• Title/Summary/Keyword: 첨가 시기

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Comparison of Vitrification and Slow Freezing for the Cryopreservation of Mouse Pronuclear Stage Embryos (생쥐 전핵기 배아 냉동보존에서 완만동결과 유리화동결의 비교)

  • Kim, Mi-Young;Lee, Yu-Il
    • Clinical and Experimental Reproductive Medicine
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    • v.34 no.2
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    • pp.117-124
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    • 2007
  • Objective: The aim of this study was to compare the efficacy of slow freezing with vitrification method for cryopreservation of mouse pronuclear stage embryos. Methods: Mouse pronuclear embryos obtained from superovulated mice and classified into 2 groups of slow freezing and vitrification. Slow freezing solution consisted of 1.5 M PROH, 0.1 M sucrose, while vitrification solution consisted of 40% ethylene glycol, 18% Ficoll and 0.5 M sucrose diluted in Dulbecco's phosphate-buffered saline supplemented with 10% SSS. Recovery and survival rates after thawing and development rates to hatching balstocyst stage were compared between two groups. Results: After freezing and thawing, recovery rate of slow freezing group was 93.8%, whereas vitrification group was 66.5% (p<0.01). Survival rate of recovered embryos were similar between two groups as 83.2% in slow freezing and 87.6% in vitrification. Embryo development rates to 2-cell stage after 24 hrs (77.0% vs 59.1%), 4-cell after 48 hrs (72.6% vs 53.3%), blastocyst after 96 hrs (53.1% vs 40.1%) of thawing were significantly higher in vitrification group than those of slow freezing group, respectively. Conclusion: The vitrification method may provide better developmental competence of frozen-thawed embryos than that of slow freezing method for cryopreservation of mouse pronuclear stage embryos.

Effects of Dietary Supplementation of Yeast Pichia farinosa on Performance, Intestinal Microflora, and Fecal $NH_{3}$ Emission in Laying Hens (효모 Pichia farinosa의 첨가 급여가 산란계의 생산능력, 장내미생물 변화 및 분의 암모니아 발생량에 미치는 영향)

  • 김상호;박수영;유동조;이상진;최철환;성창근;류경선
    • Korean Journal of Poultry Science
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    • v.29 no.3
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    • pp.205-211
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    • 2002
  • A feeding trial was conducted to study the effects of a live yeast, Pichia farinosa culture(PF), on the production performance and intestinal microflora in laying hens. One hundred and sixty ISA Brown layers, 21 weeks of age, were randomly allotted to four dietary treatments, with four replicates per treatment. Dietary treatments consisted of four levels (0, 0.1, 0.3, and 0.5%) of PF added to a com-soybean meal based diet. Egg production, egg weight, feed intake and fred conversion ratio(FCR) were measured. Egg qualifies were examined at 25th and 29th weeks of age. A metabolism trial was conducted following the feeding trial, during which intestinal microflora, nutrient digestibility and fecal NH3 gas emission were measured. Egg production of birds fed 0.1 and 0.3% PF were significantly higher than those from birds fed 0 and 0.5% PF(P<0.05). Daily egg mass of 0.3% PF increased significantly compared to that of 0% PF. There was no difference in egg weight among all treatments. Feed conversion ratio was significantly improved as the PF level increased. No significant difference was found in eggshell quality and Haugh unit at both 25 and 29 weeks of age. Viable count of ileal Lactobacillus spp. increased significantly as the rf level increased. However, the total number of yeast and anaerobes in ileum were similar among all treatments. Cecal Lactobacillus spp. and yeast counts showed no difference among all treatments. Fecal NH$_{3}$ gas emission of layers fed PF decreased significantly by the PF supplementation. From the result of this experiment, it could be concluded that dietary supplementation of the live yeast Pichia farinosa improves the laying performance and decreases the fecal ammonia gas emission.

Influence of medium addition and agitation on the production of embryos in isolated microspore culture of hot pepper (Capsicum annuum L.) (고추의 소포자 배양 시 배지 첨가와 진탕이 배의 생산에 미치는 영향)

  • An, Dong-Ju;Park, Eun-Joon;Kim, Moon-Za
    • Journal of Plant Biotechnology
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    • v.38 no.1
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    • pp.30-41
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    • 2011
  • The influences of the agitation as well as the addition of medium during culture on the production of embryos were invested in isolated microspore culture of hot pepper (Capsicum annuum L.). When the culture medium was added during initial liquid culture step of liquid-double layer culture, the embryo yield and quality greatly increased. The most effective time point for medium addition was 5 days after the culture commenced. On the other hand, the effect of medium addition at later double layer culture step in liquid-double layer culture on the embryo production was less compared to that of medium addition during the initial liquid culture step. Agitating the culture for 1 week during later double layer culture step in liquid-double layer culture effectively increased the production of normal cotyledonary embryos. In the case of liquid culture, agitating the culture for 1 week from 7 days after the culture commenced was also effective for embryo development. However, when the total agitation time was longer (2 to 3 weeks) during liquid-double layer culture or liquid culture, the embryos developed abnormally in both cases. The normal cotyledonary embryos obtained in this study successfully developed to plants when transferred to regeneration media. These regenerated plants were either diploid or haploid, and there was a difference in the number of chloroplasts between guard cells of diploid and haploid. These results can be used as an important data for developing an efficient microspore culture system with high quality embryo production in hot pepper.

Inhibitory Effects of Extracts from Albizzia julibrissin on Gonadal Maturation in a Medaka (Oryzias latipes) (자귀나무 추출물이 송사리(Oryzias latipes) 생식소 성숙에 대한 억제효과)

  • Lee, Eun-Hee;Oh, Sang-Pil;Kim, Myong-Hee;Kim, Kwang-Hyun;Hong, Sang-Hoon;Han, Chang-Hee
    • Journal of Life Science
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    • v.23 no.3
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    • pp.333-340
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    • 2013
  • This study was performed to determine whether stem bark extract containing saponin of Albizzia julibrissin in the diet influences gonadal maturation and spawning in medaka (Oryzias latipes). The crude extraction containing saponin (HaBC) was partially purified from n-BuOH extraction of A. julibrissin stem bark by Diaion HP-20, Silica gel and Sephadex LH-20 chromatographies. We fed diets supplemented with HaBC to medaka. We then studied the effects of the HaBC supplement on the suppression of gonadal maturation and spawning in female medaka that were reared in aquaria with recirculation systems. In the experiment with immature female medaka, the periods of initiation of gonadal maturation and spawning were delayed in the fish that were fed diets supplemented with at least HaBC 20 mg/g-feed. In the experiment with mature female medaka, the fish that were fed diets supplemented with at least HaBC 20 mg/g-feed had lower GSIs than the control diet group did. The results showed that the growth of the immature medaka was not correlated with the amount of supplementation of HaBC in the diet. However, the condition factors (CF) in the medaka that were fed diets supplemented with at least HaBC 20 mg/g-feed were higher than in the medaka fed on the control diet. We concluded that extracts containing saponin from the stem bark of A. julibrissin have the potential to inhibit gonadal maturation in female medaka, but they did not act as growth stimulation. Further studies are required to determine the mechanism of the action.

Development to Hatching Blastocysts and Cell Allocation to the Inner Cell Mass and Trophectoderm of Pig In Vitro Embryos as Affected by Amino Acids and Serum (아미노산과 혈청이 돼지 수정란의 내부세포괴와 영양배엽세포로의 발달과 부화에 미치는 영향)

  • Uhm, Sang-Jun;Kim, Eun-Young;Kim, Myo-Kyung;Yi, Bong-Kyung;Lee, Hyeon-Sook;Kim, Te-Oan;Yoon, San-Hyun;Park, Se-Pill;Chung, Kil-Saeng;Lim, Jin-Ho
    • Clinical and Experimental Reproductive Medicine
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    • v.24 no.2
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    • pp.241-251
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    • 1997
  • 체외성숙과 수정된 돼지 난자의 체외발달능이 체외배발생 배양액인 NCSU 배양액에 0.4% BSA, 10% 혈청 혹은 아미노산 (2% BME 아미노산 용액과 1% MEM 아미노산 용액)을 첨가함으로서 조사되었다. 본 실험에 공시된 난자는 체외수정 추 30시간 (2-세포기)혹은 48 시간 ($2{\sim}4$-세포기)에 회수하였다. 실험I에서 0.4% BSA가 첨가된 NCSU 배양액에서 2-세포기 난자들의 배양경과시간에 따른 발달능을 조사한 결과, 배양 후 72 시간 (체외수정 후 102 시간)에 상실배기와 배반포기 배가 나타났으며, 배양 후 120 시간째 (체외수정 후 150 시간)에도 팽창된 배반포기 배까지만 발달하였다. 실험II는 체외수정 후 48 시간의 분할된 ($2{\sim}8$-세포기) 난자들의 핵과 외관적 분할구와의 수적 차이를 조사한 결과, $2{\sim}4$-세포기보다는 5-세포기 이상에서 핵과 분할구의 조화에 차이가 많았다. 실험III에서는 $2{\sim}4$-세포기 난자들을 배양후 5일째의 배반포들의 투명대의 두께, 난자 크기 그리고 inner cell mass (ICM)과 trophectoderm (TE)의 세포 배열을 조사한 결과, 난자의 크기가 커짐에 따라서 투명대가 얇아지고 전체 세포수가 증가하였지만, ICM의 비율은 차이가 없었다. 실험IV에서는 BSA, 혈청 혹은 아미노산이 첨가 혹은 무첨가된 배양액내에서 $2{\sim}4$-세포기 난자들의 배반포 후 부화능력을 조사한 결과, 모든 군에 있는 난자들은 팽창된 배반포기 배까지 발달할 수 있었던 반면, 난자의 부화는 아미노산 혹은 혈청이 포함된 배양액에서만 일어났다. 더우기 상실배기와 배반포기 시기에 혈청의 첨가는 부화 배반포기 배의 발달을 현저히 증가시켰다. 또한 아미노산과 혈청의 영향을 받은 팽창 배반포기 배는 얇은 투명대, 팽창된 난자의 크기 그리고 ICM과 전체 세포수의 증가를 보였다. 이상의 결과로 미루어 볼때, 배양액내에 대한 아미노산과 혈청의 첨가는 돼지 배반포기 배의 부화를 유도할 수 있다고 보며, 더우기 이들 요소들은 투명대의 두께, 난자의 크기 그리고 ICM과 전체 세포수에 영향을 미친다.

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Effect of SeaR gene on virginiamycins production in Streptomyces virginiae (희소방선균 SeaR 유전자가 Streptomyces virginiae의 virginiamycins 생산에 미치는 영향)

  • Ryu, Jae-Ki;Kim, Hyun-Kyung;Kim, Byung-Won;Kim, Dong-Chan;Lee, Hyeong-Seon
    • Korean Journal of Microbiology
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    • v.51 no.3
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    • pp.256-262
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    • 2015
  • In order to study the effect of the receptor protein (SeaR), which is isolated from Saccharopolyspora erythraea, we introduced the SeaR gene to Streptomyces virginiae as host strains. An effective transformation procedure for S. virginiae was established based on transconjugation by Escherichia coli ET12567/pUZ8002 with a ${\varphi}C31$-derived integration vector, pSET152, which contained int, oriT, attP, and $ermEp^{\ast}$ (erythromycin promotor). Therefore, the pEV615 was introduced into S. virginiae by conjugation and integrated at the attB locus in the chromosome of the recipients by the ${\varphi}C31$ integrase (int) function. Transformants of S. virginiae containing the SeaR gene were confirmed by PCR and transcriptional expression of the SeaR gene in the transformants was analyzed by RT-PCR, respectively. And, we examined the production time of virginiamycins in the culture media of both the transformants and the wild type. The production time of virginiamycins in the wild type and transformants was the same. When 100 ng/ml of synthetic $VB-C_6$ was added to the state of 6 or 8 hour cultivation of wild type and transformants, respectively, the virginiamycins production was induced, meaning that the virginiamycins production in the wild type was detected 2 h early than transformants. From these results, SeaR expression was also affected to virginiamycins production in transformants derived from S. virginiae. In this study, we showed that the SeaR protein worked as a repressor in transformants.

Effects of Increasing Moisture Content with or without Supplementing Inoculant (Lactobacillus plantarum) in TMR on Its Feed Value (TMR 제조 시 수분 함량 및 발효제 (Lactobacillus plantarum) 첨가유무가 사료가치에 미치는 영향)

  • Ki, Kwang-Seok;Kim, Hyeon-Shup;Lee, Hyun-June;Lee, Wang-Shik;Baek, Kwang-Soo;Kim, Sang-Bum;Lim, Keun-Bal;Jeo, Joon-Mo;Kim, Yong-Kook
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.27 no.3
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    • pp.197-208
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    • 2007
  • This study was carried out to investigate effects of increasing moisture content with or without supplementing inoculant (Lactobacillus plantarum) in TMR (total mixed ration) on its feed value. In case of exposing TMR to air, the lower the moisture level of TMR was, the less its apparent condition was changed. The time of spreading of molds tended to be faster in TMR with the higher moisture level. And also the odor was influenced by moisture content and inoculant supplement that is, sour odor was smelled from 24 hour after exposing TMR containing 35% and 50% moisture to air, but TMR supplemented with inoculant had sweet odor. The inner temperature of TMR containing 35% and 50% moisture without inoculant tended to increase continually after the lapse of 6 hours when the TMR was exposed to air. The inner temperature of TMR containing 35% moisture with inoculant tended to increase dramatically after the lapse of 48 hours when exposed to air, but that of TMR containing 50% moisture with inoculant tended to increase after 6 hours. The pH of TMR containing 15% was consistent regardless of exposing time to air, but that of TMR containing 35% and 50% moisture considerably increased after 12 and 24 hours, respectively. The concentration of $NH_3-N$ of TMR supplemented with inoculant was increased from 6 hours after exposure to air, while that or TMR without inoculant increased from 12 hours. Nutrient content or TMR tended to be increased with the increase of exposing time to air and storage time under sealed condition.

Anti-Obesity Effect and Fermentation Characteristics of American Preferred Kimchi Added to Garcinia Cambogia Extracts (Hydroxy Citric Acid) (가르시니아 캄보지아 추출물(Hydroxy Citric Acid) 첨가 미국인 선호 김치의 발효특성 및 항비만 효과)

  • Yang, Yu-Jin;Ahn, In-Sook;Han, Ji-Sook
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.34 no.6
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    • pp.776-783
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    • 2005
  • To develop functional kimchi which had anti-obesity effect, garcinia cambogia extract containing $51.46\%$ hydroxy citric acid (HCA) was used as a sub-ingredient of American preferred kimchi (APK). The APK added to garcinia cambogia extracts of 0.5$\%,\;1.0\%,\;1.5\%\;and\;2.0\%$ were prepared, and fermentation characteristics and anti-obesity effect of those kimchi were investigated. The pH of APK added to garcinia cambogia extracts (APKH) was low as the amount of garcinia cambogia extract increased at initial stage of fermentation but the pH of those kimchi showed similar values after optimum ripened stage. The number of Lactobacillus sp. and Leuconostoc sp. were small as the amount of garcinia cambogia extract increased while the period was delayed that the number of Lactobacillus sp. and Leuconostoc sp. attained to maximum. In Hunter's color values of APKH, lightness and redness decreased as the amount of garcinia cambogia extracts increased while yellowness increased. Sensory scores in overall acceptance, taste and texture of APKH evaluated by Americans as sensory panels were similar until the addition amount of garcinia cambogia extract was $1.5\%$, therefore the garcinia cambogia extract of $1.5\%$ was determined as tile amount adding to APK. The secretions of glycerol and leptin as a key signalling factor for anti-obesity effect were examined in APK and APKH added to garcinia cambogia extract of $1.5\%$. There were no significant differences in the glycerol secretion when adipocytes were treated with APK and APKH extracts. However, leptin secretion in the adipocytes treated with APKH extract was significantly decreased compared to that of control (p<0.05).

Synthesis of Modified Polyetherimide and Toughening of Epoxy Resin (변성 폴리에테르이미드의 합성과 이를 이용한 에폭시 수지의 강인화)

  • Lee Sin Duk;Ahn Byung Hyun;Lee Kwang Gi;Kim Won Ho
    • Polymer(Korea)
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    • v.29 no.3
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    • pp.231-236
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    • 2005
  • Amino terminated polyetherimide (AP-PEI) has been synthesized using 2,2'-bis [4-(3,4-dicarboxyphenoxy)-phenyl]propane dianhydride (BPADA) and m-phenylenediamine. Polyetherimide containing pendant carboxy group (CP-PEI) has also been synthesized by the reaction of BPADA, m-phenylenediamine and 3,5-diaminobenzoic acid. The modified PEIs were used as toughening agent for diglycidyl ether of bisphenol-A epoxy resin which was cured with nadic methyl anhydride (NMA). Thermal properties, fracture toughness ($K_{IC}$) and solvent resistance of toughened epoxy resin were measured. The $K_{IC}$ of epoxy resin containing 20 phr of AT-PEI was 2.88$MPa{\cdot}m^{0.5}$ without sacrificing thermal properties. The $K_{IC}$ of epoxy resin which contained 20 phr of CP-PEI was 2.82$MPa{\cdot}m^{0.5}$.

Studies on the Changes in Activities of ALPase, ACPase, ATPase and Synthesis of Volutin Granules upon Phosphate Concentration in Saccharomyces uvarum (Saccharomyces uvarum의 인삼염첨가배양에 따른 ALPase, ACPase, ATPase 활성도와 volutin과립의 함량변화)

  • 이기성;최영길
    • Korean Journal of Microbiology
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    • v.23 no.2
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    • pp.84-89
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    • 1985
  • The effect of exogenous phosphate supply on the regulation of phosphate metabolism was investegated during catabolic repression and catabolic derepression in yeast (Saccharomyces uvarum). As the results, when sugar was supplimented in cells cultivated under phosphate free, the growith rate was low but it was capable of cell division. Polyphosphate "B" was accumulated highly in proportion to amount of phosphate added to the medium. Without regard to phosphate supply of the medium, the total amount of polyphospgate was almost similar, although each polyphosphate was turned over. Activities of all phosphatases remained continuousoy high in the cells cultivated in the phosphate free medium. Especially under catabolic repression, the function of polyphosphate system was shown to compensate the ATP/ADP system as phosphate donor, energy source and regulator.

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