• Title/Summary/Keyword: in vitro preservation

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Effect of Short-term High $CO_2$ on Growth of Botrytis cinerea (고농도 이산화탄소의 단기 처리가 Botrytis cinerea 생장에 미치는 영향)

  • 최정희;정문철;임정호
    • Food Science and Preservation
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    • v.11 no.2
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    • pp.246-249
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    • 2004
  • This study was conducted to evaluate the inhibition efficacy of high CO$_2$ atmosphere (35, 60, and 100% ${\times}$ 24, 48, and 72 h) on growth of Botrytis cinerea in vitro in order to offer sterilizing method of horticultural crops including peach fruits. Botrytis cinerea was isolated from a naturally infected peach fruits. Growth of the fungus at 25$^{\circ}C$ declined with increased CO$_2$ concentration and treatment duration. Especially, 100% CO$_2$ provided completely inhibition effect of growth of the fungus for 72 h. After removal of high CO$_2$ condition, however, the fungus showed normal growth speed. The growth of fungus at low temperature was completely inhibited temporarily by short-term 100% CO$_2$ treatment, but resumed right after transferring to normal atmosphere at 25$^{\circ}C$.

In-Vitro Fertilization and Culture of Pig Oocytes Matured In-Vitro by Liquid Boar Sperm Stored at 4$^{\circ}C$

  • Kim, M. Y.;Y. J. Yi;Y. J. Chang;Park, C. S.
    • Proceedings of the KSAR Conference
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    • 2003.06a
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    • pp.63-63
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    • 2003
  • This study was carried out to investigate the effects of liquid boar sperm stored at 4$^{\circ}C$ on sperm motility, normal acrosome, and in-vitro fertilization and culture of pig oocytes matured in-vitro. The sperm-rich fraction (30~60 ml) of ejaculate was collected into an insulated vacuum bottle. Semen was slowly cooled to room temperature (20~23$^{\circ}C$) by 2 h after collection. Semen was transferred into 15 ml tubes, centrifuged at room temperature for 10 min at 800$\times$g, and the supernatant solution was poured off. The concentrated sperm was resuspended with 5 ml of lactose, egg yolk and N-acetyl-D-glucosamine (LEN) diluent to provide 1.0$\times$10$^{9}$ sperm/ml at room temperature. The resuspended semen was cooled in a refrigerator to 4$^{\circ}C$ and preserved for 5 days to examine sperm motility and normal acrosome. The medium used for oocyte maturation was modified tissue culture medium (TCM) 199. After about 22 h of culture, oocytes were cultured without cysteamine and hormones for 22 h at 38.5$^{\circ}C$, 5% $CO_2$ in air. Oocytes were inseminated with liquid boar sperm stored at 4$^{\circ}C$ for 2 days after collection. Oocytes were coincubated for 6 h in 500 ${mu}ell$ mTBM fertilization media with 0.2, 1, 5 and 10$\times$10$^{6}$ /ml sperm concentration, respectively. At 6 h after IVF, oocytes were transferred into 500 ${mu}ell$ Hepes-buffered NCSU-23 culture medium for further culture of 6, 48 and 144 h. There were significant differences in sperm motility and normal acrosome among preservation days and incubation times, respectively. The rates of sperm penetration and polyspermy were higher in 5 and 10$\times$10$^{6}$ sperm/ml than in 0.2 and 1$\times$10$^{6}$ sperm/ml. Male pronuclear formation was lower in 0.2$\times$10$^{6}$ sperm/ml than in 1, 5 and 10$\times$10$^{6}$ sperm/ml. Mean numbers of sperm in penetrated oocyte were highest in 10$\times$10$^{6}$ sperm/ml compared with other sperm concentrations. The rate of blastocysts from the cleaved oocytes (2~4 cell stage) was highest in 1$\times$10$^{6}$ sperm/ml compared with other sperm concentrations. In conclusion, we found out that liquid boar sperm stored at 4$^{\circ}C$ could be used for in-vitro fertilization of pig oocytes matured in-vitro. Also, we recommend 1$\times$10$^{6}$ ml sperm concentration for in-vitro fertilization of pig oocytes.

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Propagation of Endangered Species, Daphne pseudomezereum var. koreana via in vitro Bud Culture (멸종위기종 두메닥나무(Daphne pseudomezereum var. koreana)의 줄기 기내배양을 통한 식물체 생산)

  • Chu, Yerin;Park, Sanghee;Cheong, Eun Ju
    • Journal of Korean Society of Forest Science
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    • v.109 no.2
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    • pp.189-194
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    • 2020
  • Daphne pseudomezereum var. koreana is native to Korea and is distributedin Kangwon-do, Jeollabuk do, and Gyeongsang-do. This economically valuable species has experienced a dramatic decrease in natural habitat due to climate change and is difficult to cultivate. In this study, we investigate a mass propagation method for D. pseudomezereum through in vitro culture and genetic resource preservation.WPM medium was better than the MS medium for shoot growth. As a result, we compared the shoot number and length of apical (W/AP) and non-apical shoots (W0/AP) with BA and GA3 treatments in WPM medium. Their shoots and length grew well in both BA 8ìM + GA38ìM-treated apical shoot and without-apical shoot. NAA did not effectively induce rooting of the in vitro plantlet.

Production, Nutritional Quality and In vitro Methane Production from Andropogon gayanus Grass Harvested at Different Maturities and Preserved as Hay or Silage

  • Ribeiro, G.O. Jr.;Teixeira, A.M.;Velasco, F.O.;Faria, W.G. Junior;Pereira, L.G.R.;Chaves, A.V.;Goncalves, L.C.;McAllister, T.A.
    • Asian-Australasian Journal of Animal Sciences
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    • v.27 no.3
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    • pp.330-341
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    • 2014
  • Andropogon gayanus is an important grass due to its high biomass production, drought tolerance and favorable growth on low fertility acidic soils. Currently, there is little research on the impact of growth stage on the nutritional quality or the degree of $CH_4$ production that may arise from this forage during ruminal fermentation. The objectives of this study were to determine the effects of regrowth stage of A. gayanus on its chemical composition, in vitro production of gas and CH4, as well as in vitro dry matter (DM) digestibility when grown under tropical Brazilian conditions and conserved as hay or as silage. The nutritional value of A. gayanus grass declined with increasing maturity; however digestible DM yield linearly increased. After 112 d of regrowth, A. gayanus produced higher quality silage (higher lactate and lower pH and butyrate content) and higher DM yield. However, the low levels of crude protein at this time would make protein supplementation a necessity for proper rumen fermentation. No differences in $CH_4$ kinetic parameters were found with advancing maturity or preservation method (hay or silage).

Changes on the Components of Free Polysaccharide from Cell Wall of Persimmon Fruit by Treatments of Cell Wall Degrading Enzymes (세포벽분해효소의 처리에 따른 감과실의 세포벽 유리 다당류의 변화)

  • 신승렬;김미현
    • Food Science and Preservation
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    • v.2 no.1
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    • pp.173-183
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    • 1995
  • This paper was carried out to investigate changes in chromatograms of polysacctatides and soluble pectins on Sephadex G-50 and non-cellulosic neutral sugars of polysaccharides isolated from cell wall of persimmon fruits treated with polygalacturonase and $\beta$-galactosidase in vitro. The chromatogram pattern of soluble pectins extracted from cell wall treated with $\beta$-galactosidase on Sephacryl S-500 column were similar to those of untreatment, but contents of soluble pectins treated with $\beta$-galactosidase were different from those of untreatment. The patterns of chromatograms In soluble pectins extracted from cell wall treated with polygalacturonase were more complex and lower molecular polymer than those of other cell wall-degrading enzyme treatments. Non-cellulosic neutral sugar of polysaccharides in fraction I of soluble material treated with polygalacturonase was rhamnose, those in fraction II were similar to those in fraction III and contents of arabinose, xylose and glucose were higher than contents of other non-cellulosic neutral sugars. Non-cellulosic neutral sugars of polysaccharides in fraction I in soluble material by $\beta$-galactosidase treatment were rhamnose, arabinose, galactose and mannose. Content of glucose of polysaccharides in fraction II was higher than that in fraction I . Non-cellulosic neutral sugars treated with mixed enzyme were rhamnose, fucose, arabinose, xylose, mannose, galactose and glucose. Compositions of non-cellulosic neutral sugars of polysaccharides in fraction I were similar to those in fraction II and III.

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Antioxidant Activity of Flavonoids in Plant Origin Food (식물성 식품에 존재하는 Flavonoids의 항산화 활성)

  • 김건희;최미희
    • Food Science and Preservation
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    • v.6 no.1
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    • pp.121-135
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    • 1999
  • Effective synthetic antioxidants such as butylated hydroxyanisole(BHA) and butylated hydroxytoluene(BHT) have been widely used in the food industry, but they are suspected to be toxic and carcinogenic effects. Therefore, the development of safely available natural antioxidants such as ascorbic acid, ${\alpha}$-tocopherol, ${\beta}$-carotene, flavonoids and selenium is essential. In particular, flavonoids, 2-phenyl-benzo-${\alpha}$-pyrones, are polyphenolic compounds that occur ubiquitously in food of plant origin. flavonoids occur in foods generally as O-glycosides with sugars bound usually at the C\ulcorner position. And variations in their heterocyclic ring gibes rise to flavones, flavonols, flavanones, flavanols, catechins, anthocyanidins, chalcone and isoflavones. Vegetables, fruits, and beverages are the main dietary sources of the flavonols, primarily as quercetin, kaempferol, and myricetin and the corresponding flavones, apigenin and luteolin. These flavonoids have biological activity such as antioxidant, anti-inflammatory, antithrombotic, antimutagenic, anticarcimogenic antiallergic and antimicrobial activity effects in vitro and in vivo. Flavonoids posses strong antioxidant activities acting as oxygen radicals scavenger, metal chelators and enzyme inhibitor. The antioxidant activity of flavonoids is determined by their molecular structure and more specially, by the position and degree of hydroxylation of the ring structure. All flavonoids with the 3`, 4`-dihydroxy(ortho-dihydroxy) posses marked antioxidant activity. And antioxidant activity increases with the number of hydroxyl groups substituted on the A-and B-rings. There is as yet no certainty about the effect of the presence of a double bond between C\ulcorner and C\ulcorner on the antioxidant activity of flavonoids.

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Genotoxicological Safety of the Two gamma-Irradiated Herbs ; Houttuynia cordata Thunberg and Kycium Chinense Miller (감마선조사 생약재 (어성초, 구기자)의 안전성에 관한 유전독학성적 평가)

  • Jo, Sung-Kee;Yu, Young-Beob;Oh, Heon;Kwak, Youn-Gil;Byun, Myung-Woo
    • Food Science and Preservation
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    • v.7 no.1
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    • pp.68-73
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    • 2000
  • These experiments were performed to investigate the safety of two herbs-Houttuynia cordata Thunberg and Lycium chinese Miller-irradiated with gamma-rays in respect of genotoxicity. Water extracts from the 10 kGy gamma-irradiated herbs were examined in two short -term in vitro tests ; (1) Salmonella typhimurium reversion assay (Ames test) in strain TA 98 and Ta100 and (2) Micronuclues test on clutured Chinese hamster ovary(CHO) cells. No mutagenicity was detected in the two assays with or without metabolic activation . From these results , the safety of the herbs irradiated with gamma-rays at practical doses could be revealed in further tests of genotoxicity in vivo, chronic and reproductive toxicity.

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A Simple Confocal Microscopy-based Method for Assessing Sperm Movement

  • Kim, Sung Woo;Kim, Min Su;Kim, Chan-Lan;Hwang, In-Sul;Jeon, Ik Soo
    • Development and Reproduction
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    • v.21 no.3
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    • pp.229-235
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    • 2017
  • In the field of reproductive medicine, assessment of sperm motility is a key factor for achieving successful artificial insemination, in vitro fertilization, or intracellular sperm injection. In this study, the motility of boar sperms was estimated using real-time imaging via confocal microscopy. To confirm this confocal imaging method, flagellar beats and whiplash-like movement angles were compared between fresh and low-temperature-preserved ($17^{\circ}C$ for 24 h) porcine sperms. Low-temperature preservation reduced the number of flagellar beats from $11.0{\pm}2.3beats/s$ (fresh sperm) to $5.7{\pm}1.8beats/s$ and increased the flagellar bending angle from $19.8^{\circ}{\pm}13.8^{\circ}$ (fresh) to $30.6^{\circ}{\pm}15.6^{\circ}$. These data suggest that sperm activity can be assessed using confocal microscopy. The observed motility patterns could be used to develop a sperm evaluation index and automated confocal microscopic sperm motility analysis techniques.

Evaluation of the mechanical properties and clinical efficacy of biphasic calcium phosphate-added collagen membrane in ridge preservation

  • Lee, Jung-Tae;Lee, Yoonsub;Lee, Dajung;Choi, Yusang;Park, Jinyoung;Kim, Sungtae
    • Journal of Periodontal and Implant Science
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    • v.50 no.4
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    • pp.238-250
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    • 2020
  • Purpose: This study aimed to evaluate the biocompatibility and the mechanical properties of ultraviolet (UV) cross-linked and biphasic calcium phosphate (BCP)-added collagen membranes and to compare the clinical results of ridge preservation to those obtained using chemically cross-linked collagen membranes. Methods: The study comprised an in vitro test and a clinical trial for membrane evaluation. BCP-added collagen membranes with UV cross-linking were prepared. In the in vitro test, scanning electron microscopy, a collagenase assay, and a tensile strength test were performed. The clinical trial involved 14 patients undergoing a ridge preservation procedure. All participants were randomly divided into the test group, which received UV cross-linked membranes (n=7), and the control group, which received chemically cross-linked membranes (n=7). BCP bone substitutes were used for both the test group and the control group. Cone-beam computed tomography (CBCT) scans were performed and alginate impressions were taken 1 week and 3 months after surgery. The casts were scanned via an optical scanner to measure the volumetric changes. The results were analyzed using the nonparametric Mann-Whitney U test. Results: The fastest degradation rate was found in the collagen membranes without the addition of BCP. The highest enzyme resistance and the highest tensile strength were found when the collagen-to-BCP ratio was 1:1. There was no significant difference in dimensional changes in the 3-dimensional modeling or CBCT scans between the test and control groups in the clinical trial (P>0.05). Conclusions: The addition of BCP and UV cross-linking improved the biocompatibility and the mechanical strength of the membranes. Within the limits of the clinical trial, the sites grafted using BCP in combination with UV cross-linked and BCP-added collagen membranes (test group) did not show any statistically significant difference in terms of dimensional change compared with the control group.

Protective effect of STAR of STAR series on CCl4 induced acute hepatotoxicity by regulation of reactive oxygen species (활성산소종의 조절을 통한 음료 '별의별간'의 급성간독성 보호효과)

  • Chang, Bo Yoon;Oh, Jun Seok;Han, Ji Hye;Kim, Da Eun;Hong, Jae Heoi;Kim, Sung Yeon
    • Food Science and Preservation
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    • v.23 no.2
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    • pp.275-282
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    • 2016
  • STAR of STAR (SS 01-04) is a series of drinks that consist of various extracts obtained from Coriolus versicolor, Artemisia capillaris Thunb., Hovenia dulcis, Acanthopanax sessiliflorus, Lycium chinense, Citrus reticulata, Saururus chinensis, Pueraria lobata, Pyrus pyrifolia, and Oenanthe javanica. A purpose of this study was to investigate the hepatoprotective effect of SS 01-04. Antioxidant activity of the drinks was evaluated by conducting a hydroxyl radical-scavenging assay. Cytotoxicity and hepatoprotective potential were determined using HepG2 cells in vitro, while protective effects against acute hepatotoxicity was evaluated in vivo. The antioxidant activity of the SS 01-04 at concentration of 100 and 250 mg/mL was similar to that of $50{\mu}M$ vitamin C. tert-Butyl hydroperoxide (tBHP)-induced production of reactive oxygen species (ROS) was blocked by SS 01, 03 and 04 in a dose-dependent manner. Treatment with SS 04 significantly lowered the serum levels of alanine aminotransferase and aspartate aminotransferase in an animal model of carbon tetrachloride $(CCl_4)-induced$ hepatotoxicity (p<0.05). In addition, SS04 increased glutathione level while decreased malondialdehyde level in the liver considerably (p<0.05). It also inhibited the $CCl_4-induced$ increase in the levels of triglyceride and cholesterol in serum and the liver. These findings indicated that SS 01-04 possessed antioxidant activity and protect against ROS. In particular, SS 04 is potentially highly beneficial in treating liver damage as it scavenges reactive free radicals and boosts the endogenous antioxidant system.