• Title/Summary/Keyword: in vitro condition

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Studies on the Survival Rates after Ultrarapidly Frozen-Thawing of Porcine Embryos (돼지 수정난의 초급속 동결 융해후의 생존성에 관한 연구)

  • 김상근;이봉구
    • Korean Journal of Animal Reproduction
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    • v.16 no.2
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    • pp.125-131
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    • 1992
  • This study was carried out to investigate the effects of concentration and equilibration time of cryoprotective agents on the survival rate of slowly and ultrarapidly frozen porcine embryos. The porcine embryos following dehydration by cryoprotective agents and a various concentration of sucrose were directly plunged into liquid nitrogen and thawed in 38$^{\circ}C$ water bath. Survival rate was defined as development rat to the morula and blastocyst stage after in vitro culture or by FDA test. The results are summarized as follows : 1. The survival rates of porcine embryos after ultrarapid frozen-thawing in the freezing medium of 0.25M sucrose added 2.0, 2.5, 3.0, 3.5, or 4.0M glycerol was 65.3, 61.8, 64.3, 59.4 or 39.4%, respectively. Addition of 0.25M sucrose into the freezing medium containing 2.0M glycerol showed higher survival rate than those of 2.5~4.0M glycerol. 2. The survival rates of porcine embryos after ultraradpid frozen-thawing in the freezing medium of 0.25M sucroese added 2.0, 2.5, 3.0, 3.5 or 4.0M DMSO was 65.6, 67.6, 68.6, 60.6 or 23.6%, respectively. However, addition of 0.25M sucrose into the freezing medium containing 3.0M DMSO showed higher survival rate than those of 2.0, 2.5, 3.5 or 4.0M DMSO. 3. The survival rates of porcine embryos after ultrapid frozen-thawing in the freezing medium of 0.25M sucrose added 2.0, 2.5, 3.0, 3.5 or 4.0M propanediol was 63.2, 60.3, 62.1, 52.3 or 24.3%, respectively. Addition of 0.25M sucroese into the freezing medium containing 2.0M propanediol showed higher survival rate than those of 2.5~4.0M glycerol. 4. The survival rates of porcine embryos after ultrarapid frozen-thawing the freezing medium of 2.0M glycerol added 0.10, 0.25, 0.50 or 0.75M sucrose was 61.8, 70.8, 67.6 or 52.2%, respectively. Addition of 2.0M glycerol into the freezing medium containing 0.25M sucreose showed higher survival rate than that those of 0.10, 0.50 or 0.75M sucrose. 5. The higher suvival rate of porcine embryos were attained at short period of equilibration time 92.5~5min.) in the freezing medium added 0.25M sucreose and 3.0M compared to those of 10 or 20min. equilibration time in the same condition.

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The Effect of Higenamine on the Calcium Transport of Cardiac Sarcoplasmic Reticulum (Higenamine이 심근 Sarcoplasmic Reticulum의 칼슘운반에 미치는 영향에 관한 연구)

  • Kim, Joo-Hyun;Lee, Yung-Kyoon;Kim, Hae-Won;Kim, Myung-Suk;Park, Chan-Woong;Lim, Jung-Kyoo
    • The Korean Journal of Pharmacology
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    • v.18 no.2
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    • pp.79-87
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    • 1982
  • Higenamine(dl-demethylcoclaurine, dl-1-(4-hydroxybenzyl)-6,7-dihydroxy-1,2,3,4-tetrah-ydroisoquinoline hydrochloride), which has recently been isolated from Aconite root by Drs. Kosuge and Yokota, has known to be the main cardiotonic component of the Aconite root. The present study was undertaken to investigate the effects of Higenamine on the calcium binding and release and ATPase activity of fragmented cardiac sarcoplasmic reticulum under in vitro condition. The calcium binding and release of sarcoplasmic reticulum were measured by using the double-beam spectrophotometer and the calcium sensitive dye, murexide. In the presence of $10^{-4}{\sim}5{\times}10^{-3}M$ of Higenamine, the maximal calcium binding and the initial binding rate of porcine cardiac sarcoplasmic reticulum were inhibited dose dependently by up to 43%. However, the calcium release from cardiac sarcoplasmic reticulum, which was loaded with $Ca^{++}(50{\mu}M)$, was stimulated in dose dependent manner. When incubated in the medium of 20 mM Tris-maleate(pH 7.0), 100 mM KCl, 10 mM $MgCl_2,\;0.05mM\;CaCl_2\;and\;0.014{\sim}1\;mM\;Tris-ATP\;at\;30^{\circ}C$ in the presence of Higenamine $(10^{-4}{\sim}5{\times}10^{-3}M)$, both $Ca^{++}-and\;Mg^{++}-ATPase$ of sarcoplasmic reticulum were inhibited non-competitively by Higenamine and values of $K_i$ were 4.896 mM and 6.875 mM respectively. It is suggested from the above findings that the cardiotonic effects of Higenamine might be partially explained by the inhibition of calcium binding and the stimulation of calcium release from the sarcoplasimic reticulum which may increase the free intracellular calcium that is available in the contraction of the cardiac muscle fiber.

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Introduction of two-step culture method for multiple seed bulb development from shoot tip culture of garlic (Allium sativium L.) (마늘의 경정배양에서 기내인경구 대량생산을 위한 2단계 배양의 도입)

  • Hwang, Hye-Yeon;Lee, Young-Bok
    • Journal of Plant Biotechnology
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    • v.35 no.1
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    • pp.75-80
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    • 2008
  • In vitro culture of shoot tip of garlic (Allium sativium L. cv. Seosan) was carried out to find medium condition of the induction of multiple shoots and bulbing for muliproduction of virus-free seed bulbs. For this work, tank culture was introduced. In shoot tip culture on MS solid medium the induction of multiple shoots and bulbing were better by adding 3% sucrose than 8%. Supplementation with 2mg/L 2ip and 0.2 mg/L IAA in this medium was effective. Three point three shoots including 2.7 bulbs were formed from a shoot tip after cultivation for 30 days on this medium. Bulbing of garlic in liquid culture with plastic water tank of 20L supplied air at the side of the lower part was better by adding 3% sucrose than 8% by subculture for 45 days with shoots obtained from shoot tip culture for 30 days on soid MS medium. Shoot growth was vigorous at 3% sucrose however bulb growth was more effective on the medium of 8% sucrose. Because of the effectiveness on solid medium added 3% sucrose, 2 mg/L 2ip and 0.2 mg/L IAA for initial production of multi-shoot in stem tip culture and the effectiveness in liquid culture with water tank for growth of bulbs, the method of two-step culture could be introduced for the multiple production of seed bulb of high quality. It was more desirable by supply of 0.2 mg/L BA and 0.02 mg/L NAA at tank culture time. But growth of the bulbs became poor by increasing concentration of NAA of the medium.

Establishment of Producing Conditions of Fermentation Feed for Swine (양돈용 발효사료의 발효조건 설정 연구)

  • Cho, S.B.;Kim, D.W.;Yang, S.H.;Park, K.H.;Choi, D.Y.;Yoo, Y.H.;Hwang, O.H.
    • Journal of Animal Environmental Science
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    • v.18 no.3
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    • pp.137-144
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    • 2012
  • This study was conducted to determine the effect of feed additives including probiotics, moisture and feed ingredients and the effect of fermented feed on digestibility and volatile fatty acid (VFA) level in finishing pigs. Feed was mixed with microbials including Saccharomyces, Lactobacillus, Enterococcus and Pediococcus together with different levels of probiotics, 0.25, 0.5, 1, 2, 3%. Addition of probiotics showed improved fermentation rate after 48 h incubation. To determine the optimal moisture level for fermentation, different levels of water, 30, 40, 50 and 60%, were added into the feed. Fermentation rate of feed with 40~50% moisture level was higher than that from 60% level at 60 h post-fermentation. In vitro fermentation rate of feed ingredients was analyzed by comparing VFA levels. Beet pulp and tapioca showed higher fermentation rate compare to other ingredients including canola meal or rapeseed meal. To determine the effect of administration of fermented feed In vivo, feces from finishing pigs were analyzed. Finishing pigs administrated with fermented feed showed improved digestibility and higher volatile fatty acid (VFA) level. In conclusion, results from the current study indicate that 40~50% of moisture with addition of beet pulp and tapioca in feed is optimal condition for fermentation. Furthermore, our data suggest that fermentation of feed can improve the feed quality and digestibility, thereby provide more nutrient in finishing pigs.

Probiotic Characterization of Acid- and Bile-tolerant Lactobacillus salivarius subsp. salivarius from Korean Faeces

  • Bae, H.C.;Nam, M.S.;Lee, J.Y.
    • Asian-Australasian Journal of Animal Sciences
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    • v.15 no.12
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    • pp.1798-1807
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    • 2002
  • This study was conducted to investigate lactobacillus salivarius subsp. salivarius having probiotic properties to be used as the health adjuncts with fermented milk products. Acid- and bile-tolerant lactobacillus salivarius subsp. salivarius was isolated with lactobacilli MRS broth from faeces of 80 healthy persons (infants, children and adults). It was used as a probiotic strain in fermented milk products. The pH of fermented milk decreased from pH 6.7 to 5.0 and titratable acidity increased from 0.3% to 1.0% by L. salivarius subsp. salivarius (isolation strain 20, 35, and 37), when incubated for 36 h at 37$^{\circ}C$. The number of viable cell counts of fermented milk was maximized at this incubation condition. The SDS-PAGE evidenced no significant change of casein but distinct changes of whey protein were observed by isolated L. salivarius subsp. salivarius for titratable acidity being incubated by 0.9-1.0% at 37$^{\circ}C$. All of the strains produced 83.43 to 131.96 mM of lactic acid and 5.39 to 26.85 mM of isobutyric acid in fermented products. The in vitro culture experiment was performed to evaluate ability to reduce cholesterol levels and antimicrobial activity in the growth medium. The selected L. salivarius subsp. salivarius reduced 23-38% of cholesterol content in lactobacilli MRS broth during bacterial growth for 24 h at 37$^{\circ}C$. All of the isolated L. salivarius subsp. salivarius had an excellent antibacterial activity with 15-25 mm of inhibition zone to E. coli KCTC1039, S. enteritidis KCCM3313, S. typhimurium M-15, and S. typhimurium KCCM40253 when its pH had not been adjusted. Also, all of the isolated L. salivarius subsp. salivarius had partial inhibition zone to E. coli KCTC1039, E. coli KCTC0115 and S. enteritidis KCCM3313 when it had been adjusted to pH 5.7. The selected strains were determined to have resistances of twelve antibiotic. Strains 27 and 35 among the L. salivarius subsp. salivarius showed the highest resistance to the antibiotics. These results indicated that some of the L. salivarius subsp. salivarius (strain 27 and 35) are considered as effective probiotic strains with a potential for industrial applications, but the further study is needed to establish their use as probiotics in vivo.

Effects of Hormone and Several Factors on the Regeneration and Transformation rate of Potato Cultivars Bred in Korea (국내 육성 감자의 재분화와 형질전환 효율에 미치는 생장조절제의 조성 및 몇 가지 요소)

  • Yi, Jung-Yoon;Seo, Hyo-Won;Cho, Ji-Hong;Lee, Shin-Woo;Yun, Han-Dae
    • Journal of Plant Biotechnology
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    • v.30 no.1
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    • pp.27-33
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    • 2003
  • The optimal condition of in vitro regeneration and transformation were investigated for newly bred potato varieties in Korea. Leaf and internodal stem tissues of 'Chubak', 'Namsuh', 'Jasim', 'Jopung' and 'Jowon' were used to investigate the influence of growth regulator on regeneration efficiency The effect of phenolic compound acetosyringone on gene transformation efficiency was also investigated. Leaf tissue of 'Jowon' and internodal stem tissues of 'Jopung' were showed high regeneration efficiency on M5 medium supplemented with 0.1 mg/L GA₃, 2.0 mg/L Zeatin and 0.01 mg/L NAA. The other three potato cultivars were showed low regeneration efficiency less than 25%. The effect of acetosyringone on Agrobacterium-mediated gene transformation with leaf and internodal stem tissues were noticeable. By adding the 75 μM of acetosyringone during the Agrobacterium innoculation, the transformation efficiency was increased up to 1.5∼4.0 fold compare to non-treated control. In case of 'Jowon' the transformation efficiency was 87.9% in leaf tissue and 'Jopung' was 68.4% in internodal stem tissues.

In Vitro Propagation of Lilium Asiatic Hybrid 'Hae Hwa' via the Formation of Shoot Clusters (신초 Cluster 형성에 의한 Lilium Asiatic Hybrid 'Hae Hwa'의 기내번식)

  • Han, Bong-Hee;Yu, Hee-Ju;Yae, Byeoung-Woo;Goo, Dae-Hoe
    • Journal of Plant Biotechnology
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    • v.29 no.1
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    • pp.19-23
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    • 2002
  • This experiment was conducted to micropropagate bulblets via shoot cluster formation and massproduce normal bulblets from the sections of proliferated shoot clusters in Lilium asiatic hybrid 'Hae Hwa'. The induction of shoot clusters from the culture of bulblet sections was more effective than that of bulb scales on MS medium with 1.0 mg/L BA and 0.5 mg/L IAA. Proliferation of shoot clusters from the formed shoot cluster sections was the most favorable on medium containing 5.0 mg/L BA and 0.5 mg/L IAA. The formation and the growth of bulblets from shoot cluster sections were achieved effectively on medium with 60∼90 g/L sucrose. The leaves derived from shoot clusters grew vigorously but the bulblets from shoot clusters grew very poor in 5L air-lift bioreactor culture. By the addition of 30 mL fresh liquid medium containing doulble strength MS salts, 250 g/L sucrose and 5 g/L activated charcoal after 8 weeks in the shoot cluster culture on MS medium with 5.0 mg/L BA and 0.5 mg/L IAA, the number of bulblets was increased in light condition, but the growth of bulblets was not affected by light. Bulblet production was possible with the bulblet product at 53 to 68 mg in fresh weight by liquid medium addition after the proliferation of shoot cluster.

A Case Study on Performance Analysis of Antimicrobial Copper Film Attaching to Window for Responding to COVID-19 and Others (코로나19 등 대응을 위한 "유리창 부착용 항바이러스 동필름" 성능분석 사례연구)

  • Kim, Seong Je
    • Journal of Korean Society of Disaster and Security
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    • v.14 no.1
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    • pp.23-40
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    • 2021
  • In the era of the global coronal 19 pandemic, there is a risk of cross-infection in hospitals at the stage where treatments and vaccines are currently being developed and marketed, so individuals should enhance their acquired immunity and generalize their living systems by the performance of copper ions in the social environment. In order to prevent the spread of infection, the need for anti-bacterial film and its efficacy were analyzed through anti-viral performance tests based on research and development cases of worldwide and immemorial time. he Korea Construction Research Institute (KCL) has received anti-bacterial performance certification and anti-viral test scores from the "National Approval Performance Certification Agency." At the time, NCCP 43326 Human Corona virus (BetaCoV/Korea/KCDC03/2020), which was approved by the Centers for Disease Control and Prevention, was introduced to ensure that the activity rate of infected cells was satisfied in the anti-viral performance test. Anti-proliferation measures for the Corona 19 virus require a quality clinical trial study comparing the experimental group within the glass space where the antiviral copper film is constructed with the comparator of the same condition without copper film.

Screening for Effective Organic Farming Materials for the Control of Tuber Rot of Gastrodia elata Caused by Fusarium spp. (천마 괴경썩음병 방제에 효과적인 유기농업자재 선발)

  • Kim, Chang-Su;Lee, Han-Na;Kim, Hyo-Jin;Seo, Sang-Young;Jo, Jong-Hyun;Lee, Eun-Suk;Ahn, Min Sil;Kim, Hee-Jun
    • Korean Journal of Organic Agriculture
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    • v.29 no.4
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    • pp.561-574
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    • 2021
  • This study was conducted to evaluate the control efficacy of the organic farming materials (OFMs) on tuber rot of Gastrodia elata caused by Fusarium spp. The antifungal activities in vitro as well as the suppressive effect of 15 OFMs on the spore germination and germ tube growth by inoculating spore suspension on immature tubers in vivo were investigated. 7 OFMs inhibited the mycelial growth of Fusarium spp. and 7 of them were microbial agents. In the screening using immature tubers, 3 OFMs were very effective with control efficacy value of 70%. Among them, sulfur provided suppressive effect on both mycelial growth and spore germination against tuber rot of G. elata. Finally, 3 OFMs were selected to test the protective and curative effects, and all chosen OFMs significantly suppressed disease incidence when applied in the preventive action, in comparison with the curative action. Especially, sulfur and Bacillus subtilis gave excellent protective control efficacy with control values of 93.2% and 86.9%, respectively, whereas its curative control effect was relatively low (73.3%, 60.2%). On the other hand, the preventive and curative effects of citronella + paraffin oil + ethyl alcohol were 73.3% and 67.0%, respectively. This study suggests that tuber rot of G. elata can be controlled by some OFMs in the rain shelter greenhouse under continuous cultivation condition and protective treatment is more important and efficient.

The Effects of Swimming Training on Lymphocyte Proliferation and ROS Production in Spleen Lymphocytes of BALB/c Mice (규칙적인 수영훈련이 마우스 비장세포의 ROS생성과 림프구 증식에 미치는 영향)

  • Kwak, Yi-Sub;Park, Jeon-Han;Kim, Se-Jong;Jang, Yun-Soo;Lee, Bong-Ki
    • IMMUNE NETWORK
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    • v.2 no.2
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    • pp.96-101
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    • 2002
  • Background: Aerobic training can be defined as any physical exercise that increases the heart rate and enhances the body's intake of oxygen long enough to benefit the condition of body. Running, cycling, and swimming are examples of aerobic activities. This type of exercise optimises immune functions. Recently several experimental findings suggested that the regular swimming training increase immune response, but there have been very few reports which compare warm water exercise with cold water exercise in spleen lymphocytes. Methods: This study was designed to examine the effects of regular swimming training on Index, the number of lymphocytes, proliferative activity and production of reactive oxygen species (ROS) by splenocytes in BALB/c mice. Thirty six mice (6 week old) were performed 10 weeks of regular swimming training and they were divided into 6 groups according to the regular swimming training (CRG: control resting group, CEG: control exercise group, WRG: warm water trained resting group, WEG: warm water trained exercise group, CORG: cold water trained resting group, COEG: cold water exercise group). Analytical items were weight change, spleen index, the number of lymphocytes, proliferative activity and production of ROS. All data were expressed as mean and standard deviation by using SPSS package program (ver. 10.0). Results: The swimming training significantly decreased body weight, and increased spleen index, the number of lymphocytes and proliferative activity in the presence or absence of Con A and LPS added conditions. For the WRG and CORG, the quantity of ROS from splenocytes was higher than CRG, whereas, ROS by spleen lymphocytes was lower following 90 min acute exercise stress. Conclusion: These results suggested that the swimming training not only increases the number of lymphocytes but also increases proliferative activity by splenocytes in vitro.