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Effect of Cryoprotectant Kinds and Cell Stages on the Viability of Mouse Embryos Cryopreserved by OPP Vitrification (동결보호제의 종류 및 배발달단계가 OPP Vitrification 동결보존시 생쥐수정란의 생존성에 미치는 영향)

  • 공일근;조성균;조성근
    • Korean Journal of Animal Reproduction
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    • v.23 no.1
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    • pp.85-92
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    • 1999
  • This study was designed to determine effect of cryoprotectant kinds and cell stages on OPP vitrification method in mouse embryos. The freezing speed, cryoprotectants and cell stage could affect of embryo viability following various vitrification methods. The vitrification solution used were consisting of 40% (v/v) ethylene glycol, 18% (w/v) Ficoll, 0.3 M sucrose solution in holding medium (D-PBS supplemented with 5% FCS: HM) (EFS) or 16.5% ethylene glycol , 16.5% dimethyl sulfoxide, 0.5 M sucrose in HM (EDS). The embryos were collected from oviduct at 18 h after hCG injection and then washed and cultured in mHTF medium until use. In experiment 1, the blastocysts were vitrified by OPP straw to determine the optimal vitrification solution of EFS or EDS. The post-thaw survival rates at re-expanded stage rates were significantly different between EFS and EDS (95.0 vs 100%), but at hatching stage was not different between EFS and EDS (90.0 vs 95.0%). respectively. In experiment 2, zygotes, 2-, 4-cell, morula and blastocysts were vitrified by OPP method to determine the acceptable of early stage embryos. The development rates to expanded blastocyst in zygote (70.0%) were significantly lower rather than those in 2-, 4- 8-cell, compacted morula or blastocyst (89.7, 90.0, 92.8, 97.6 or 97.5%), respectively. However, the cell number of post-thaw developed to expanded blastocyst in blastocyst and control blastocyst stage (39.6$\pm$2.81, 35.7$\pm$2.98) were significanty higher than those in zygote, 2-, 4-, 8-cell, compacted morula (29.8$\pm$3.21, 31.3$\pm$3.83, 29.3$\pm$3.58, 28.9$\pm$3.21 or 30.8$\pm$2.93). In experiment 3, the zygotes were exposed in VSl for 1, 2, and 3 min to the optimal exposed time. The cleavage rates (91.6, 88.5, 88.9%) and develop mental rates to blastocyst (83.3, 74.3 and 69.4%) depends on the exposed time in VSl were not significantly different among 1, 2, or 3 min, respectively. The cell number also were not significantly different among exposed time in VS1. respectively. These results indicate that OPP method could be useful for vitrification either EFS or EDS vitrification solution. The post-thaw survival rates at zygote were significantly lower than those at 2-, 4-, 8-cell, morula or blastocyst, respectively. The zygote stage were more sensitive rather than late stage embryos. The exposing time in VS1 for 1 min was better than that for 2 or 3 min, even it was not significantly different. The OPP vitrification method could be useful of mouse embryos either with EFS or EDS vitrification solution.

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Hyaluronic acid 및 Tocopherol 첨가가 돼지 난포란 체외 발달율에 미치는 영향

  • 이향혼;임기순;최선호;이창현;김광식;이연근;박진기;장원경;송해범
    • Proceedings of the KSAR Conference
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    • 2001.03a
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    • pp.27-27
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    • 2001
  • 본 연구는 돼지 난포란의 체외 성숙시 hyaluronic acid (HA) 및 Tocopherol첨가가 돼지 수정란의 배발달에 미치는 효과를 검토하기 위하여 실시하였다. 돼지 난포란은 0.1% PVA, 3.05mM D-Glucose, 0.91mM sodium pyruvate, 0.57mM cysteine, 0.5$\mu\textrm{g}$/$m\ell$ LH, 0.5$\mu\textrm{g}$/$m\ell$ FSH and 10ng/$m\ell$ EGF가 첨가된 TCM-199에서 HA와 Tocopherol을 농도별로 첨가하여 42~44시간 배양함으로써 체외성숙을 유도하였다. HA의 효과를 보기 위해 체외성숙시 0.1% HA가 첨가된 처리구에서 배발달율은 25.6%로 대조구 16.7% 보다 높았다.(Table Omitted). 또한 수정시 HA를 0, 0.02, 0.2, 2.0 로 처리한 결과 8.7%, 19.4%, 11.1%, 0%로 대조구(8.7%)에 비해 0.02% 처리구(19.4%)에서 배발달율이 가장 높았다. 수정후 control, TCM-199, TCM-199+0.1% HA가 첨가된 배양액에 washing해주었을 때 발달율은 TCM-199+0.1% HA 첨가구에서 배발달율(35.6%)이 유의적으로 높게 나타났다. Tocopherol을 0, 50, 100, 200mM로 처리한 결과 수정된 난자중 배발달율은 32.1%, 40.0%, 35.7%, 39.6%로 처리구에서 높았으며, 성숙배양 후반기 22h. 동안 Tocopherol 첨가한 후 수정전 HA로 washing한 결과 Tocopherol 무첨가구에서는 차이를 보이지 않았으나, Tocopherol 첨가구에서는 HA처리구에서(19.0%)에서 무처리구보다(14.3%) 높은 결과를 나타내었다. 따라서 배양액내에 HA, Tocopherol를 첨가함으로써 미성숙 난포란의 수정율 및 발달율을 높일수 있을 것으로 생각된다.

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Dormancy of Ginseng Seed as Influenced by Temperature and Gibberellic Acid (인삼종자의 휴면기간단축에 미치는 온도 및 지베레린의 영향)

  • Jong-Chul Lee
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.31 no.2
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    • pp.220-225
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    • 1986
  • This study was conducted to know the effects of temperature and gibberellic acid on the dormancy block-ing of ginseng seeds at different embryo growth stage. Optimum temperature for embryo growth appeared to be I soc at the beginning stage of post ripening (up to dehiscent time), 1$0^{\circ}C$ at middle stage (for 30 days after dehiscence) and 5$^{\circ}C$ at last stage (between 30 and 92 days after dehiscence). And optimum temperature for dehiscence is about 17$^{\circ}C$, also the optimum temperature for dehiscence is higher than that of embryo growth. Germination of ginseng seed with full grown embryo was accelerated at high temperature (25-3$0^{\circ}C$). Germina-tion percentage was 80% at 105th date after dehiscence under 5$^{\circ}C$, 28% at 147th date under 1$0^{\circ}C$, but no germination under the over 15$^{\circ}C$. Gibberellic acid increased the dehiscent rate, whereas the gibberellic acid treatment may not be substituted for effect of low temperature on the germination. Low temperature may be reguired to finish the embryo growth in thickness.

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Plant Regeneration via Secondary Somatic Embryogenesis and Acclimatization in Panax ginseng (장뇌삼의 2차 체세포배 발생을 통한 식물체 유도 및 순화)

  • Lee, Su-Gwang;Kim, Ji-Hee;Kang, Ho-Duck
    • Journal of Korean Society of Forest Science
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    • v.97 no.1
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    • pp.127-133
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    • 2008
  • This study was conducted to establish the optimal condition for plant regeneration and acclimatization from somatic embryos of Panax ginseng. Cotyledon segments of Panax ginseng produced primary and secondary somatic embryos when cultured on MS and WPM media supplemented with 7% sucrose. To induce plantlet conversion, cotyledonary somatic embryos were cultured on WPM solid medium with $GA_3$ at various concentrations (1~30 mg/L) for 4 weeks. Plantlets were transferred to 1/2 WPM solid medium with $GA_3$ at various concentrations (0~5 mg/L) and 0.5% activated charcoal for shoot and root elongations. Elongated plantlets further developed into well-developed leaf and root system on 1/3 SH medium with 0.5% activated charcoal under ventilation condition for 5 months. The highest survival rate to soil was 75% when plantlets were regenerated on 1/3 SH medium without sucrose under ventilation condition.

In Vitro Propagation of Zanthoxylum piperitum DC. - II. Effect of $NH_4NO_3, KNO_3$ and Casein hydrolysate on Somatic Embryogenesis- (초피나무 (Zanthoxylum piperitum DC) 의 기내증식 - II. $NH_4NO_3, KNO_3$ Casein hydrolysate의 기내 부정배 발생효과 - ( In Vitro Propagation of Zanthoxylum piperitum DC. - II. Effect of NH4NO3 , KNO3 and Casein hydrolysate on Somatic Embryogenesis - ))

  • 송원섭
    • Korean Journal of Plant Resources
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    • v.8 no.2
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    • pp.153-157
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    • 1995
  • Embryogenic callus induces from shoot tip and leaf segment of Zanthoxylum piperitum for producing somatic embryogenesis and plant regeneration were cultured in vitro on Murashige and Tucker's(MT) medium treated with casein hydrolysate $NH_4NO_3$, $KNO_3$ and plant growth regulator. The most effective somatic embryogensis was observed in the medium added by two fold $NH_4NO_3$(3300mg/l)+2. 4-D 0.1mg/l and $KNO_3$(3800mg/l)+2.4-D 0.1mg/l. Also, MT medium supplemented with casein hydrolysate 700mg/l added by 2, 4-D 0.1mg/l were effective in obtainingn somatic embryos from embryogenic callus The effect ofm MT medium supplemented with casein hydrolysate without 2, 4-D was lower than that with (3300mg/l) 2, 4-D for the formation of somatic embryos.

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Effect of Oxygen Concentrations with Superoxide Dismutase on In Vitro Maturation of Porcine Follicular Oocytes and In Vitro Development of Porcine IVM/IVF Embryos (산소농도 및 Superoxide Dismutase가 돼지 난포란의 체외성숙과 배발달에 미치는 영향)

  • 한만희;이규승
    • Journal of Embryo Transfer
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    • v.16 no.3
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    • pp.173-182
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    • 2001
  • The present study was carried out to examine the effect of superoxide dismutase (SOD) on in vitro maturation (IVM) of porcine follicular oocytes and oxygen concentration with SOD on in vitro development (IVD) of porcine IVM/IVF embryos. The results were summarized as follows : 1. The rates of nuclear maturation, penetrated oocytes, polyspermic oocytes and mean numbers of the penetrated sperm were not different in the NCSU-23 maturation media with 0, 100, 500 and 1,000 units/ml SOD. However. the pronucleus formation rates were significantly lower in oocytes matured with addition groups than those of no addition groups of SOD (P>0.05). 2. The rates of blastocyst formation and total cell numbers of blastocyst at day 7 after in vitro fertilization were significantly lower in addition groups than those of the no addition groups of SOD (P>0.05). 3. The rates of blastocyst formation at day 7 after in vitro fertilization were higher in the NCSU-23 culture medium with 100 units/ml SOD than in those cultured with 0, 500 and 1,000 units/ml SOD under the 5% and 20% $O_2$concentrations. However, no differences was found in the total cell numbers of blastocyst among the treatments. In conclusion, these results suggested that the addition of SOD was not adequate for porcine oocyte maturation and further development, also the rates of blastocyst formation and total cell numbers of blastocyst at day 7 of porcine IVM/IVF embryos were not different in the NCSU-23 culture medium under the 5% and 20% $O_2$concentrations.

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Impact of Awareness and Educational Experiences on Cardiopulmonary Resuscitation in the Ability to Execute of Cardiopulmonary Resuscitation among Korean Adults (한국 성인에서 심폐소생술에 대한 인지, 교육경험이 그 시행능력에 미치는 영향)

  • Lee, Jae-Kwang;Kim, Jeongwoo;Kim, Kunil;Kim, Keunhyung;Kim, Dongphil;Kim, Yuri;Moon, Seonggeun;Min, Byungju;Yu, Hwayoung;Lee, Chealim;Jeong, Wonyoung;Han, Changhun;Huh, Inho;Park, Jung Hee;Lee, Moo-Sik
    • Journal of agricultural medicine and community health
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    • v.43 no.4
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    • pp.234-249
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    • 2018
  • This study was performed to identify the impact of awareness and educational experiences on cardiopulmonary resuscitation in the ability to execute of cardiopulmonary resuscitation among Korean adults. This study used original data of 2014 Community Health Data Survey. 228,712 participants in this survey were resident in South Korea who is aged 19 or older on July 2014. Participants in this survey were sampled an average of 900 residents(target error ${\pm}3percent$) per community health center of Korea. Data were analyzed by using R 3.1.3 employing chi-squared test, fisher's exact analysis, and logistic regression analysis. Ability to execute CPR was significantly higher in males(3.34 time), higher the education level (1.61 times), the white color occupation (1.14 times), the higher the income level (1.07 times), the higher the education level (0.91 times), non-hypertensive patients (1.12 times), non-diabetic patients (1.16 times), non-dyslipidemic patients (0.86 times), non-stroke patients (0.30 times), CPR education experience group (3.25 times), CPR experience group with manikin-based training (4.30 times), higher subjective health status (1.08 times, 1.16 times) respectively. This study identified that awareness, educational experience, and mannequin-based learning experience of CPR impacted on the ability to execute CPR. Responding to education-related factors could contribute to reducing the rate of out-of-hospital acute cardiac arrest by improving the ability to execute CPR of the general public.

배 품종별 생장시기에 따른 이화학적 특성

  • Jang, Sun;Na, Chang-Soo;Eun, Jong-Bang
    • Proceedings of the Korean Society of Postharvest Science and Technology of Agricultural Products Conference
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    • 2003.04a
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    • pp.105.2-106
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    • 2003
  • 배 폐과를 효율적으로 활용하여 식품소재나 이를 이용한 기능성식품 개발을 위하여 배 품종별 생장시기에 따른 이화학적 특성을 조사하였다. 나주 지역에서 생산한 풍수, 신고, 추황 등 3품종을 시료로 하여 생장시기에 따라 배 무게의 증가를 관찰하였고 표면의 착색도는 Hunter 색차계로 L, a, b값을 측정하였으며 수분함량은 상압가열건조법으로 측정하였다. pH와 가용성 고형물은 각각 pH meter와 refractometer로, 총산은 적정법으로 측정하여 citric acid로 나타내었다. 총당은 phenol-황산법으로, 환원당의 함량은 DNS법으로 정량 하였고 총탄수화물의 함량과 전분의 함량은 산 가수 분해시킨 후 DNS법으로 측정하였다. 유기산과 유리당은 HPLC를 이용하여 정성, 정량 분석하였다. 풍수 과중의 증가는 거의 직선적으로 증가하였고 신고와 추황은 생장전기에는 급속히 증가하다가 후기에는 완만한 증가를 보였다. 풍수는 L, a, b값이 모두 약간의 증가를 보였고 신고와 추황은 a값이 각각 -1.80, 4.01에서 9.22, 9.70으로 증가하였으며 L, b값은 큰 변화를 나타내지 않았다. 과피의 수분함량은 약 61%에서 79%로 증가하였고 과육의 수분함량은 76%에서 90%로 증가하였다. 풍수와 추황의 경우 과육의 수분함량은 생장초기에 이미 생장후기와 비슷한 수분함량 비율을 갖고 있는 것을 알 수 있었다. 3품종 모두 생장초기 총산의 함량은 수확시기의 함량보다 높았고 추황의 경우 성장이 진행됨에 따라 과피와 과육의 총산의 함량은 감소하다가 다시 약간의 증가 추세를 나타내었다. 유기산은 주로 tartaric acid, malic acid, oxalic acid, citric acid 그리고 shikimic acid가 검출되었고 수확시기에 3품종에서 모두 malic acid 함량이 가장 높았고 malic acid와 citric acid의 함량이 풍수와 신고에서는 약 0.3%, 추황은 0.4% 이상으로 나타났다. 또한 3품종 모두에서 총산과 같은 추세로 성숙기의 유기산의 함량이 생장초기의 함량보다 많이 감소되었음을 알 수 있었다. 배의 가용성 고형물은 전체적으로 과실의 성장이 진행됨에 따라 증가하는 추세를 나타내었다. 풍수와 신고 과피의 총당과 환원당은 거의 같은 추세로 증가하였고 성숙이 가까워지면서 환원당이 감소하였다. 배 과육의 총당과 환원당 또한 거의 같은 추세로 증가를 하다가 수확 전 20일 혹은 30일부터 총당의 함량은 계속 증가하지만 환원당의 함량은 큰 변화가 없는 것으로 나타났다. 배 과실의 전반 성장과정에 있어서 전분함량은 감소하였고 총탄수화물의 함량은 과육에서는 증가하나 과피에서는 반대로 감소하는 경향을 보였다. 풍수의 전분함량은 최고 2.19%에서 0.23%로 감소하였고 신고에서는 0.43%로 추황에서는 0.48%로 감소하였다. 배 유리당은 fructose, glucose, sorbitol, sucrose 둥 4종류의 당이 검출되었고 3품종 모두에서 생장과정 중 비환원당인 sucrose 함량은 계속 증가하였고 fructose, glucose, sorbitol의 함량(추황의 sorbitol을 제외)은 생장이 촉진됨에 따라 증가하다가 다시 점차적으로 감소하였다. 이러한 결과는 총당과 환원당의 측정결과와 일치한 것으로 나타났다. 결론적으로 배의 성장에 따라 산 함량은 감소하였고 당 함량은 증가하였다.

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Analysis of Fine Particle Transfer and Shear Strength Increase Using PFC in Permeation Grouting (PFC를 이용한 침투그라우팅시 미세입자의 이동 및 전단강도증가 해석)

  • Lee, Wan-Ho;Lim, Heui-Dae
    • Journal of the Korean Geotechnical Society
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    • v.23 no.11
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    • pp.49-58
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    • 2007
  • Numerical experiments using a distinct element code (PFC3D) were carried out for the analysis of grout-material transfer in soil layers and also for the analysis of increase in mechanical strength after permeation grouting. For rapid analysis, up-scaling analysis in length scale was adopted, and the following observations were made from the numerical experiments. Firstly, the relative size of grout material with respect to the in situ soil particles controlled the transfer distance of the grout particles. When the size of grout particle was 0.2 to 0.25 times of the in situ soil particles, clogging of pore spaces among the in situ soil particles occurred, resulting in restricted propagation of grout particles. It was also found that there was a threshold value in the size of grout particle. Below the threshold value, the transfer distance of the grout particle did not increase with the decrease of particle size of the grout material. Secondly, the increase in cohesion and internal friction angle was observed in the numerical specimen with grouting treatment, but not with the untreated specimen.

Effects of Cytochalasin B on Parthenogenetic Development of Porcine Follicular Oocytes (돼지 난포란의 단위발생에서 Cytochalasin B의 영향)

  • Kim, Hyun-Jong;Son, Dong-Soo;Choi, Sun-Ho;Cho, Sang-Rae;Choe, Chang-Yong;Kim, Young-Gun;Han, Man-Hye;Ryu, Il-Sun;Kim, In-Cheul;Kim, Il-Hwa;Im, Kyung-Soon
    • Development and Reproduction
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    • v.9 no.1
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    • pp.49-52
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    • 2005
  • The effects of cytochalasin B was studied for the cleavage and development of in vitro matured porcine follicular oocytes. The follicular oocytes were collected from slaughtered pig ovaries and matured for 65 hours. The matured oocytes were activated by 7% ethanol(v/v) in DPBS and the activated oocytes were subjected to cytochalasin B concentrations of 2.5, 5.0 and $7.5\;{\mu}g/mL$ for 3, 5 and 7 hours, and then the treated oocytes were cultured in NCSU23 with 0.4% BSA for 7 days. The cleavage rates were not different significantly in each treatment. However, the oocytes treated with $5.0\;{\mu}g/mL$ for 5 hours yielded a significantly higher morula rate(19.7%) than oocytes treated with $2.5\;{\mu}g/mL$ for 3 and 5 hours(9.4%). The sum rate of $2.5\;{\mu}g/mL$ concentration(10.5%) by hour was also significantly lower than those of 5.0(18.0%) and $7.5\;{\mu}g/mL$ concentration(14.6%). The blastocyst rate in oocytes treated with $5.0\;{\mu}g/mL$ for 3(9.4%) and 5 hours(9.0%) was significantly higher than the rate in oocytes treated with $2.5\;{\mu}g/mL$ for 3 hours(0%). The sum rate of $5.0\;{\mu}g/mL$ concentration also significantly higher than those of 2.5 and $7.5\;{\mu}g/mL$ concentration. The results demonstrated that the treatment of oocytes with cytochalasin B of $5.0\;{\mu}g/mL$ for $3{\sim}5$ hours was the optimal concentration and duration for parthenogenetic activation and blastocyst formation of in vitro matured porcine oocytes.

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