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Effects of Different Media and Oxygen Concentrations on In Vitro Maturation and Development of Porcine Follicular Oocytes (배양액과 산소농도가 돼지난포란의 체외성숙과 배발달에 미치는 영향)

  • 천행수;한만희;김종화;박병권;이규승;서길웅
    • Reproductive and Developmental Biology
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    • v.28 no.2
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    • pp.119-126
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    • 2004
  • The present study was carried out to examine the effect of four different media (NCSU (North Carolina State University)-23, PZM (Porcine Zygotes Medium)-3, PZM-4 and TCM (Tissue Culture Medium)-l99) and two oxygen concentrations (39 , 5% $O_2$, 5% $CO_2$ and 90% $N_2$, 5% $CO_2$ in air) on in vitro production of porcine IVM/IVF embryos. The results were summarized as follows: The rates of GVBD and nuclear maturations were not significantly different (p>0.05) for 44 hours of culture with four media in two oxygen concentrations. The rates of polyspermy, penetrated sperm(s) and male and female prouclei formation were not significantly different (p>0.05). among four media in two oxygen concentrations. The cleavage rates were not significantly different (p>0.05) among four media in two oxygen concentrations. At day 7 under gas atmosphere of 5% $O_2$, 5% $CO_2$ and 90% $N_2$, the blastocyst formation was significantly higher (p<0.05) in PZM-3 (19.9$\pm$2.4) than other media. Also, NCSU-23 medium gave high rate of blastocyst formation at day 7 under gas atmosphere of 5% $CO_2$ in air (p<0.05). Based on the result of differential staining of porcine blastocyst at dat 7, inner cell mass cell and total cell numbers were not significantly different (p>0.05) among four media in two oxygen concentrations. However, the observed total cell number was higher in PZM-3 medium (36.8$\pm$6.5) than other madia. In conclusion, these results suggested that in vitro production of porcine embryos in PZM-3 medium under a gas atmosphere of 5% $O_2$, 5% $CO_2$ and 90% $N_2$ was effective on the blastocyst formation rate and total blastocyst cell number.

Effects of Antioxidants on Porcine IVM/IVF Embryos (돼지 체외수정란의 체외발육에 있어 항산화제의 효과)

  • 장현용;오진영;김종택;박춘근;정희태;김정익;이학교;최강덕;양부근
    • Reproductive and Developmental Biology
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    • v.28 no.2
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    • pp.77-82
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    • 2004
  • The purpose of this study was performed to establish the in vitro culture system of porcine in vitro maturation and in vitro fertilization(IVM/IVF) embryo. These studies was to determine the effects of antioxidants(aesculetin, taurine and melatonin) in porcine IVM/IVF embryos. In routine porcine IVM/IVF procedure, oocytes were cultured for 40∼44h incubation in NCSU 23 mediumand matured oocytes were inseminated with frozen semen. Then 2 to 8 cell embryos were removed cumulus cell and were allotted randomly to NCSU 23 containing different concentration of antioxidants in 5% $O_2$ and 5% $CO_2$ at 38.5$^{\circ}C$. Cell numbers of blastocyst were also counted using double fluorescence stain method. Aesculetin were added to NCSU 23 medium at concentration of 1 ug, 5 ug, and 10 ug, when treated with 10 ug(35.7%) of aesucletin at the rate of embryos of the morula plus blatocsyts were higher than those of any other groups (30.2%, 29.5% and 29.2%)(P<0.05). The developmental rates beyond morula stage of porcine embryos in NCSU 23 medium supplemented with taurine 0, 2.5 and 5.0 mM were 26.1%, 26.9% and 31.7%, respectively. The addition of 5.0 mM taurine was higher the developmental rate beyond morula stage than in any other groups. In NCSU 23 medium treated with melatonin 0, 1, 5 and 10 nM, the developmental rate of morula plus blastocysts were 33.3%, 39.1%, 33.3% and 27.9%, respectively. The developmental rate of morula and blascytocys treated with 1nM melatonin was higher than in any other groups(P<0.05). Cell numbers of blastocyst in NCSU 23 treated with melatonin 0, 1, 5 and 10nM were 41.0, 42.6, 39.6 and 33.0, respectively. These results indicate that aesculetin, taurine and melation can increase the developmental rate beyond the morulae and blastocysts in porcine embryos.

The Effects of Melatonin and Sodium Nitroprusside (SNP) on Development of Porcine IVM/IVF Embryos (돼지 체외수정란의 체외발육에 있어 Melatonin과 Sodium Nitroprusside(SNP) 첨가 효과)

  • 장현용;오진영;김종택;박춘근;정희태;김정익;이학교;최강덕;양부근
    • Reproductive and Developmental Biology
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    • v.28 no.2
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    • pp.83-87
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    • 2004
  • The objective of this study was performed to establish the in vitro culture system of porcine in vitro maturation and in vitro fertilization(IVM/IVF) embryo. These studies was to determine the effects of melatonin, nitric oxide donor(SNP), and the combination effects of SNP and melatonin in porcine IVM/IVF embryos. In routine porcine IVM/IVF procedure, oocytes were cultured for 40∼44h incubation, and the zygotes were cultured for 40∼44h in NCSU 23 medium. Then 2 to 8 cell embryos were removed cumulus cell and were allotted randomly to NCSU 23 containing different concentration of melatonin, SNP and SNP plus melatonin in 5% $O_2$, 5% $CO_2$ and 90% $N_2$ at 38.5$^{\circ}C$. Cell numbers of blastocyst were also counted using double fluorescence stain method. In NCSU 23 medium treated with melatonin 0, 1, 5 and 10 nM, the developmental rate of morula plus blastocysts were 33.3%, 39.1%, 33.3% and 27.9%, respectivly. This result show that the developmental rate of morula and blascytocys treated with 1 nM melatonin was higher than in any other groups(P<0.05). The developmental rates of morula plus blastocysts were 41.9% in 0 uM SNP, 25.6% in 50 uM and 28.4% in 100 uM, respectively. The developmental rate of morula plus blastocysts were decreased treated with SNP in NCSU 23. In combined effects of SNP plus melatonin (0, SNP 50 uM, SNP 50 uM plus melatonin 1 nM, SNP 50 uM plus melatonin 5 nM and SNP 50 uM plus melatonin 10 nM), the developmental rates beyond morula stage of porcine embryos were 31.3%, 34.1%, 39.5%, 29.4% and 39.5%, respectively. The addition of SNP 50 uM plus maltonin 1 nM, developmental rates of blastocyst was higher rate than in any other groups. Cell numbers of blastocyst in NCSU 23 treated with melatonin 0, 1, 5 and 10 nM were 41.0, 42.6, 39.6 and 33.0, respectively. In combined effects of SNP plus melatonin (0, SNP 50 uM, SNP 50 uM plus melatonin 1 nM , SNP 50 uM plus melatonin 5 nM and SNP 50 uM plus melatonin 10 nM), cell numbers of developed blastocyst were 36.3, 34.6, 39.0, 39.9 and 39.0, respectively. These result show that the cell numbers of blastocyst treated with 0, 1 and 5 nM melatonin were higher than in 10 nM group(P<0.05), but cell numbers of blatocyst produced by SNP plus melatonin were not significantly difference in all experimental groups.

Variation in Muscle Lipids and Fatty Acid Composition of Neutral and Phospholipids of Mudskipper (Boleophthalmus pectinirostris) during Hibernation (동면중 짱뚱어 근육유(筋肉油)의 지방질과 중성 및 인지질 조성의 변화)

  • Kim, Myung-Kon;Baek, Seung-Hwa;Choe, Sun-Nam;Kim, Jong-Bae;Park, Il-Woong
    • Applied Biological Chemistry
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    • v.41 no.7
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    • pp.533-538
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    • 1998
  • In order to gain fundamental information on the utilization of lipids as energy source in mud-skipper, muscle lipids and their fatty acid composition were investigated with respect to life cycle-maturation (Aug), before-hibernation (Nov), and after-hibernation (Apr). Crude and neutral lipid were found to decrease from 1.2, 68.3% (Aug) via 0.7, 53.8% (Nov) to 0.4, 42.6% (Apr), respectively, whilst phospholipids and glycolipids increased from 29.5 and 2.2% (Aug) to 52.1 and 5.3% (Apr), respectively. In neutral lipids, TG contents gradually decreased from 53.8% (Aug) via 33.6% (Nov) to 23.1% (Apr), while FFA and sterol contents increased from 13.5 and 14.2% (Aug) to 22.3 and 24.5% (Apr), respectively. In phospholipids, PC content decreased from 61.2% (Aug) to 50.6% (Apr), while changes in PS and PE contents, as a whole, showed the opposite trends. In neutral lipids, the levels of some fatty acids such as 16:0, 16:1, 18:0, 18:1 and 20:5 $({\omega}3)$ were analyzed to be high, with the 20 : 5 being predominant, and the levels of saturated and monoene-acids gradually decreased, while polyene-acids increased in before and after hibernation. In before hibernation, 16:0, 16:1, 18:1 mainly decreased but 18:2, 18:3 $({\omega}3)$ and most of saturated and monoene acids such as 14:0, 14:1, 16:1, 18:0, 18:1 slightly decreased in after hibernation. From these findings, it was suggested that those fatty acids decreased during hibernation were used as a energy source, particularly 16:1 and 18:1 being most preferentially used. In phospholipids, the levels of 16:0, 18:0, 20:5 $({\omega}3)$, 22:5 $({\omega}3)$ and 22:6 $({\omega}3)$ were found to be high. Throughout the life cycle, the levels of monoene-acids in phospholipids stayed constant, whilst those of 18:2, 18:3 $({\omega}3)$ and saturated acids such as 16:0, 18:0 were found to be decreased gradually in before and after hibernation, whereas those for the high degree of polyene-acids such as 20:4 $({\omega}6)$, 20:5 $({\omega}3)$, 22:5 $({\omega}3)$, 22:6 $({\omega}3)$ increased, particularly 20:4 $({\omega}6)$, 20:5 $({\omega}3)$ being most increased.

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Comparison of Rice Growth under Subtropical and Temperate Environments (아열대와 온대 기후 하에서 벼 생육 비교)

  • Park H.K.;Xu Migging;Lee K.B.;Choil W.Y.;Choil M.G.;Kim S.S.;Kim C.K.
    • Korean Journal of Agricultural and Forest Meteorology
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    • v.8 no.2
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    • pp.45-53
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    • 2006
  • The objectives of this study are to determine the primary yield components responsible for yield differences in a subtropical environment of the Hunan province China and in a temperature environment of Honam province Korea. Field experiments were conducted in a subtropical environment in Hunan province China during 2002 and in a temperate environment in Honam province Korea during 2003. Seven rice cultivars were grown under optimum crop management in each experiment field. Yield, yield components and plant dry matter were determined at maturation. The highest yield (567 kg/10a) was produced at Honam province by Jinyou 207, a Chinese cultivar, The maximum yield at Hunan province was 453 kg/10a by Sanyou 63. On the average across cultivars, Honam produced 23% greater yields than Hunan. Sink size (spikelets per $m^2$) was responsible far these yield differences. Panicle number per $m^2$ was much greater at Honam.

Functional implications of gene expression analysis from rice tonoplast intrinsic proteins during seed germination and development (벼 종자에서 액포막 aquaporin (tonoplast intrinsic protein) 유전자의 발현과 기능)

  • Huh, Sun-Mi;Lee, In-Sook;Kim, Beom-Gi;Shin, Young-Seop;Lee, Gang-Seop;Kim, Dool-Yi;Byun, Myung-Ok;Kim, Dong-Hern;Yoon, In-Sun
    • Journal of Plant Biotechnology
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    • v.37 no.4
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    • pp.517-528
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    • 2010
  • Rice seed maturation and germination involve drastic changes in water and nutrient transport, in which tonoplast aquaporins may play an important role. In the present study, gene expression profiles of 10 tonoplast intrinsic proteins (TIP) from rice were investigated by RT-PCR during seed development and germination. OsTIP3;1 and OsTIP3;2 were specifically expressed in mature seeds. Their transcript level rapidly decreased after onset of seed germination and gene expression was induced by ABA treatment. In contrast, expression of OsTIP2;1 and OsTIP4;3 was not seed specific as transcripts were found in vegetative tissues as well. Their respective transcript levels decreased at an early stage of seed development, whereas they increased at a later stage of seed germination and elongation of embryonic roots and shoots. When seed germination was inhibited by various stress conditions and ABA, expression of OsTIP2;1 and OsTIP4;3 was completely suppressed. In contrast, the expression level of OsTIP2;2 rapidly increased after seed imbibition and the transcript level was maintained under conditions inhibiting seed germination. These results implicate that tissue specific and developmental transcriptional regulation of OsTIPs in rice seeds depends on their specific function. In addition, OsTIPs can be discriminated by different potential phosphorylation and methylation sites in their protein structures. OsTIP3;1 and OsTIP3;2 possess unique phosphorylation signatures at their N-terminal domain, loop B and loop E, respectively. OsTIP2;1 and OsTIP4;3 have a potential methylation site at their Nterminal domain. This suggests that activity of specific tonoplast aquaporins may be regulated by post-translational modification as well as by transcriptional control.

Effect of Different Light Emitting Diode (LED) Lights on the Growth Characteristics and the Phytochemical Production of Strawberry Fruits during Cultivation (파장별 LED광이 딸기의 생장 특성과 생리 활성 물질 형성에 미치는 효과)

  • Choi, Hyo Gil;Kwon, Joon Kook;Moon, Byoung Yong;Kang, Nam Jun;Park, Kyoung Sub;Cho, Myeong Whan;Kim, Young Cheol
    • Horticultural Science & Technology
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    • v.31 no.1
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    • pp.56-64
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    • 2013
  • Recent unusual weather due to global warming causes shortage of daily sunlight and constitutes one of the primary reasons for agricultural damages. LED light sources are frequently utilized to compensate for the shortage of sunlight in greenhouse agriculture. The present study is aimed at evaluating formations of phytochemicals as well as growth characteristics of mature strawberry fruits ('Daewang' cultivar) during cultivation in a closed growth chamber equipped with artificial LED light as a sole light source. Each LED light of blue (448 nm), red (634 and 661 nm) or mixed blue plus red (blue:red = 3:7) was separately supplied and the intensity of each light was adjusted to $200{\pm}1{\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}$ at plant level with a photoperiod consisted of 16 hours light and 8 hours darkness. Strawberries grown under mixed LED light of blue and red wavelengths showed a higher production of fruits than those grown under other LED treatments. Fructose, one of the free sugars, increased in mixed LED light-grown fruits. Anthocyanin contents were elevated remarkably in the mixed LED light-grown fruits compared with those in other LED treatments. Contrastingly, contents of total phenolics and flavonoids were not of much different from one another among the fruits treated with various LED lights. On the other hand, ripening of strawberry fruits was found to be faster when grown under blue LED light compared with other LED treatments. Moreover, antioxidant activities of blue or red LED light-grown fruits, respectively, were significantly higher than those of mixed LED light-grown fruits. We suggest that when daylight is in shortage during cultivation in a greenhouse, supplementation of sunlight with LED light, which is composed of blue and red wavelengths, could be useful for the enhancement of productivity as well as of free sugar content in strawberry fruits. In addition, for the strawberry culture in the plant factory, selective adoption of LED light wavelength would be required to accomplish the purpose of controlling fruit maturation time as well as of enhancing contents of sugars and antioxidants of fruits.

Inhibitory effects of persimmon (Diospyros kaki Thumb.) against diet-induced hypertriglyceridemia/hypercholesterolemia in rats (떫은감 (Diospyros kaki Thumb.)이 흰쥐의 식이성 이상지질혈증에 미치는 영향)

  • Ahn, Youngsook;Regu, Geberea manuel Meron;Oh, Eun Kyoung;Kwon, Oran
    • Journal of Nutrition and Health
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    • v.50 no.3
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    • pp.225-235
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    • 2017
  • Purpose: This study aimed to investigate the potential of freeze-dried persimmon powder (Diospyros kaki Thumb.) to protect against dyslipidemia induced by a high-fat/cholesterol diet (HFD) in a rat model. Methods: Fifty Wistar rats were randomly divided into five groups: normal control (NC), high-fat/cholesterol control (HC), tannin in HFD (HT, 1% of diet), immature persimmon in HFD (HI, 7% of diet), and mature persimmon in HFD (HM, 7% of diet). Tannin was used as a positive control. Biochemical, molecular, and histopathological changes were observed in the blood and liver. Results: We confirmed that a high fat/cholesterol diet successfully induced dyslipidemia, which was characterized by significantly altered lipid profiles in the plasma and liver. However, oxidized low-density lipoprotein levels, histopathological damage in the liver, and hepatic triglyceride levels were significantly reduced in all HT, HI, and HM groups compared to those in the HF group. In contrast, plasma apolipoprotein B level was significantly reduced only in the HT and HM groups, whereas reduction of the LDL-C level was detected only in the HI group. Although HF-induced sterol regulatory element-binding protein (SREBP) gene expression was significantly reduced in all treated groups, downstream gene expression levels varied among the different groups; significant reduction of fatty acid synthase (FAS) and 3-hydroxy-3-methylglutaryl-CoA (HMGCR) gene expression was detected only in the HI group, whereas cholesterol $7{\alpha}$-hydroxylase (CYP7A1) gene expression was significantly elevated only in the HM group. Conclusion: Taken together, the data suggest that protection of LDL oxidation and hepatic lipogenesis might be, at least partly, attributed to tannin in persimmons. However, the identified mechanisms varied up to the maturation stage of persimmon. In the case of immature persimmon, modulation of FAS and HMGCR gene expression was prominent, whereas in the case of mature persimmon, modulation of CYP7A1 gene expression was prominent.

Histological Study on the Reproductive Cycle of Potamocorbula amurensis (Bivalvia: Corbulidae) (계화도조개 (Potamocorbula amurensis)의 생식주기에 관한 조직학적 연구)

  • LEE Ju Ha
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.32 no.5
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    • pp.629-636
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    • 1999
  • Gonadal development, gametogenesis, reproductive cycle, egg-diameter and composition, condition factor, and the first sexual maturity of the clam, Potamocorbula amurensis were investigated by histological observation. Samples were collected monthly from the tidal flat of Moonpo, Puan-gun, Chollabuk-do, west coast of Korea from November 1996 to October 1997. P. amurensis is dioecious and oviparous. The gonads were composed of a number of gametogenic follicles. The oogonia and fully ripe oocytes were $9\~12\mu$m and $50\~60\mu$m in diameter, respectively. Each of the spermatogenic follicle formed stratified layers composed of spermatogonia, spermatocytes spermatids, and spermatozoa in groups on the follicular wall. The reproductive cycle of P. amurensis could be classified into five successive stages: early active, late active, ripe, partially spawned, and recovery. Spawning occurred twice a year from May to July and from September to October, the main spawning seasons also appeared twice a year between May and June, and in October when the water temperatures reached above $18^{\circ}C$. The monthly changes in the condition factor were closely related with the reproductive cycle. Minimum size for the sexual maturation of female and male were 8.1 mm in shell length. There were two patterns for the gametogenesis: 1. After spawning, the undischarged ripe oocytes and spermatozoa in the follicles were degenerated and absorbed, but in part, the existing follicles were not contracted significantly and then they took part in new gametogenesis within one or two months (especially, in summer). 2. After spawning, each follicle was contracted, thereafter gametogenesis again occurred in newly formed follicles.

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Fruit and Wine Quality on Maturing Time of Korea New Grape Cultivar 'Doonuri' (국내 육성 포도 '두누리' 품종의 성숙 시기별 과실특성과 포도주 품질특성)

  • Chang, Eun Ha;Jeong, Sung Min;Noh, Jung Ho;Park, Kyo Sun;Lim, Byung Sun
    • Food Science and Preservation
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    • v.19 no.6
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    • pp.882-892
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    • 2012
  • This study was conducted to investigate several indicators of fruit maturity, and expert sensory evaluation was done to determine the most appropriate maturity time for the new Korea grape cultivar 'doonuri' for high quality winemaking. The grape component changed dramatically during ripening, after veraison. Considerable drops in berry firmness (<0.60 kgf/$cm^2$) and titratable acidity (<0.66%) were found at the ripening stage while the soluble solid content significantly increased in the ripening process. Most of the organic acids contained were tartaric and malic acid. The malic acid content was about twice the tartaric acid content in the fruit-growing period but was less than the tartaric acid content in the maturity period. The total anthocyanin and polyphenol contents increased sharply after veraison. In particular, the total anthocyanin and polyphenol contents reached their maximum levels when the acidity was 0.55%, and after that, there was almost no change. Correlations between anthocyanin accumulation and several factors were found in sugar 0.7811 (p<0.05), pH 0.9315 (p<0.05), and Brix/acid ratio 0.9409 (p<0.05). $Brix{\times}pH^2$ and brix/acid ratio were used as indicators of the proper maturation of the grapes when the acidity was 0.53 to 0.55%, and at sugar 17 Brix. When surveyed, the quality characteristics and sensory evaluation of the wine made using the latest harvested grapes showed the most reliable sourness. The color, aroma, and overall harmony of the wine, however, were evaluated to be the best when the wine acidity was about 0.60% or when the grape acidity was below 0.55%.