• Title/Summary/Keyword: Cold shock treatment

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Comparative Study of Growth and Gonad Maturation in Diploid and Triploid Marine Medaka, Oryzias dancena

  • Park, In-Seok;Gil, Hyun Woo;Lee, Tae Ho;Nam, Yoon Kwon;Kim, Dong Soo
    • 한국발생생물학회지:발생과생식
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    • 제20권4호
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    • pp.305-314
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    • 2016
  • The marine medaka, Oryzias dancena is a suitable sample as a laboratory animal because it has a small size and clearly distinguishes between female and male. Data on the growth and maturity of the diploid and triploid sea cucurbit species suitable for laboratory animals are very useful for studying other species. Triploidy was induced in the marine medaka by cold shock treatment ($0^{\circ}C$) of fertilized eggs for 45 min, applied two minutes after fertilization. The diploid and triploid male fish were larger than their female counterparts (P<0.05), and the concentrations of thyroid stimulating hormone (TSH) and thyroxine (T4) were higher in the induced triploids over 1 year (P<0.05). In both the diploid and tri-ploid groups the concentrations of TSH and T4 were higher in the male fish than in the females (P<0.05), while the testo-sterone and estradiol-$17{\beta}$ concentrations in the induced triploids were lower than in the diploids (P<0.05). The gonadosomatic index (GSI) of the triploid fish was lower than that for the diploids, and the GSI for females in each ploidy group were higher than that for the males. For both groups the GSI was highest at 4 months of age, and decreased thereafter to 12 months. Analysis of the gonads of one-year-old triploid fish suggested that the induction of triploidy probably causes sterility in this species; this effect was more apparent in females than in males.

느타리의 기내 자실체 형성 및 그 유도조건에 관한 연구 (Laboratory-scale fruiting body formation of Pleurotus ostreatus using the petri dish culture)

  • 조중호;추교선;김범기;공원식;유영복;이승재;조봉금;이창수
    • 한국버섯학회지
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    • 제2권1호
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    • pp.15-20
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    • 2004
  • 국내에서 많이 재배되는 버섯 중의 하나인 느타리 P. ostreatus는 먹물버섯이나 치마버섯과는 달리 실험실 수준에서의 자실체 발생이 힘든 것으로 알려져 있다. 본 연구에서는 실험실에서 느타리버섯을 샤레를 이용하여 전 발생과정을 유도하기 위한 방법을 검토하였다. 샤레상의 배양조건은 프라스틱 샤레 ($60{\times}15mm$)의 감자배지에서 균사를 접종한 후 빛이 없는 상태에서 배양한 뒤에, 균사의 환기상태, 균사표면의 상처, 빛 그리고 저온충격 등의 여러 환경요인들이 원기 및 자실체 형성에 미치는 영향을 검토하였다. 느타리의 최초 자실체 형성은 접종 이후 3주 내에 얻을 수도 있었으며, 균사접종 이후 10주 동안에 자실체의 형성은 균주에 따라서 76%에서 97%의 높은 빈도로 유도될 수 있었다. 위와 같이 샤레상에서 자실체를 형성할 수 있었으며, 정상적인 자실체의 성장을 위해서 빛은 필수적이며, 환기도 필요하였다. 또한 균사의 상처 처리가 원기, 자실체 및 포자의 형성에 미치는 영향이 균주에 따라서 크게 차이가 났으며, 같은 균주라 하더라도 발생단계별 그 반응의 차이도 크다는 사실을 확인하였다. 이들 자실체에서 수집된 담자포자는 발아가 가능하였다. 샤레상의 완전한 자실체 형성 방법은 느타리버섯의 생육주기를 단축시킬 수 있을 뿐만 아니라, 다양한 재배조건의 시료 획득과 한개의 배양 용기 내에서 버섯 발생의 전 단계를 관찰하고 분석하는데 유용할 수 있다.

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산화스트레스에 노출된 정자의 생존성 및 운동성에 있어서 커큐민의 이중효과 (Dual effect of curcumin on viability and motility of bovine sperm exposed to oxidative stress)

  • 화정석;김은진;류지현;;박창윤;최창용;강다원
    • 한국수정란이식학회지
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    • 제31권3호
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    • pp.299-305
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    • 2016
  • Although cryopreservation of sperm is routinely used for clinical requirement, it has some problems, such as high generation of reactive oxygen species (ROS) and cold-shock. To reduce the detrimental damage in sperm, anti-oxidants were added to cryoprotectant for sperm. Curcumin is one of anti-oxidants, which are added in cryoprotectants. However, recent studies have demonstrated that curcumin decreases sperm viability and motility. This study was performed to identify the effect of curcumin on hydrogen peroxide ($H_2O_2$)-exposed bovine sperm, which were cryopreserved-thawed. In $H_2O_2$-exposed bovine sperm, reactive oxygen species (ROS) were significantly reduced by treatment with curcumin in a dose-dependent manner (p < 0.05). Among tested concentrations of curcumin (1 to $50{\mu}M$), 30 and $50{\mu}M$ curcumin showed anti-oxidant effect on $H_2O_2$-induced ROS generation. On the other hand, combination of 30 or $50{\mu}M$ curcumin with anti-oxidant $H_2O_2$ increased the percentage of apoptotic sperm compared to only $H_2O_2$ treatment. Sperm viability was also decreased in the combination of 30 or $50{\mu}M$ curcumin with $H_2O_2$ as judged by FDA/PI staining. $H_2O_2$-induced decrease in sperm progressive motility was recovered by treatment with $1{\mu}M$ curcumin. These results show that high concentration of curcumin has anti-oxidant effect, but it has also cytotoxic effect on bovine sperm. Sperm viability and motility might be more affected by cytotoxic signals of curcumin compared to antioxidant signals.

미꾸라지(Misgurnus mizolepis)의 웅성발생성 처녀생식: II. 웅성발생성 이배체 유도 및 생존능력 평가 (Intraspecific Androgenesis in Mud Loach (Misgurnus mizolepis): II. Diploid Restoration and Viability Assessment)

  • 남윤권;방인철;노충환;조영선;김동수
    • 한국양식학회지
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    • 제19권4호
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    • pp.281-287
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    • 2006
  • 우리나라 주요 담수어종인 미꾸라지(Misgurnus mizolepis)를 어류 염색체 조작 모델로 개발하기 위한 연구의 일환으로 동일 종내 웅성발생성 이배체 유도 조건을 개발하고 유도된 웅성발생성 이배체의 생존능력을 평가하였다. 자외선(UV; $10,800\;ergs/mm^2$)을 이용하여 난자의 유전물질을 불활성화시키고 인공수정 28분 후(제1난할 중기)에 온도처리(고온 자극 $40.5\;^{\circ}C$ 120초 후 저온 자극 $1\;^{\circ}C$ 45분)를 통해 제1난할을 억제하였다. 본 온도처리를 통해 생존력 있는 웅성발생성 이배체들이 유도되었고(평균 부화율, 26.9%), 부화 후 1주일째를 기준으로 평균 웅성발생성 이배체 수율은 최초 처리난의 7%내외로 나타났다. 그러나 웅성발생성 이배체 유도 효율은 cytoplasmic donor의 질에 크게 영향을 받음이 관찰되어 사용한 암컷 친어별로 큰 편차가 관찰되었다. Flow cytometry 분석에 의해 생존력 있는 웅성발생성 개체들의 이배체 복원이 확인되었으며 아울러 형질전환 표지를 이용하여 동형접합성 개체 유도의 가능성이 확인되었다. 웅성발생성 이배체들은 일반대조군 이배체에 비해 낮은 생존능력을 보였으며 특히 부화 후 1개월까지 유의적으로 낮은 생존율을 나타내었다. 그러나 후기 생존능력은 일반 이배체군과 유사하였다.

비생물학적 스트레스 관련 벼 Ac/Ds 삽입 변이체의 선발 및 유전자 발현 분석 (Selection of (Ac/Ds) insertion mutant lines by abiotic stress and analysis of gene expression pattern of rice (Oryza sativar L.))

  • 정유진;박슬아;안병옥;윤도원;지현소;이강섭;박용환;서석철;백형진;이명철
    • Journal of Plant Biotechnology
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    • 제35권4호
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    • pp.307-316
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    • 2008
  • 식물에서 전이인자를 이용한 삽입 변이체의 유전자 기능분석 연구가 최근 가장 활발하게 이루어지고 있다. 본 연구에서는 동진벼의 Ac/Ds 삽입 변이체인 F2 세대 30,000 계통을 이용하여 고염과 저온에 민감한 계통과 내성이 있는 계통을 대량 스크리닝을 통해 선발하였다. 첫 번째 스크리닝에서 선발한 212 계통을 Southern blot 분석을 통해 Ds의 삽입여부 및 copy 수를 꽉인하고 표현형과 비교하여 고염과 저온에서 총 19 계통을 선발하였고, 이 중 copy 수가 하나인 계통은 13 계통이었다. 선발한 계통을 FSTs 분석을 통해 Ds의 삽입위치 및 knock-out유전자를 확인하고 염기서열 정보를 이용하여 벼 전체 염기서열 정보와 상동성 비교분석 결과 세포의 신호전달 과정과 조절 관여하는 유전자 그룹인 transpoter, protease family protein and apical meristem family protein, 삼투압조절에 관여하는 유전자 그룹인 heat shock potein, O-methyltransferase, glyceraldehyde-3-phosphate dehydrogenase and drought stress Induce protein 그리고 식물의 소포유통(vesicle trafficking)에 관여하는 유전자 SYP 5 family protein로 구분할 수 있었다. 선발된 19개 유전자의 발현 분석을 위해 9종류 비생물학적 스트레스 하에서 RT-PCR을 수행한 결과 이들 knock-out 유전자는 비생물학적 스트레스에 각각 다른 발현 패턴을 보였다. 이 연구의 결과는 삽입 변이체를 통한 유전자의 기능분석에 있어서 비생물학적인 스트레스의 응답 반응계에 관여하는 유전자를 연구하는데 유용할 것이라고 생각된다.

Methanol을 이용한 개 정액 동결시 융해후 양호한 활력 및 생존율을 나타내는 정액 처리 조건 (Semen Treatment to Maintain Good Quality of Post-thaw Motility and Viability of Canine Spermatozoa Frozen Using Methanol)

  • 김용준;박영재;김병진;유일정
    • 한국임상수의학회지
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    • 제11권2호
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    • pp.545-552
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    • 1994
  • This experiment was carried out to investigate the renditions to maintain good post-thaw motility and viability of canine spermatozoa when the semen was frozen using methanol. The semen from two male dogs which had been proven to be fertile in the previous one year was treated with different compositions of semen diluent and was frozen at different freezing temperatures, When canine semen was frozen at-2$0^{\circ}C$, -6$0^{\circ}C$, or -8$0^{\circ}C$, the spermatozoa frozen and stored at -2$0^{\circ}C$ showed very low post-thaw motility and viability from day 2 to 7 and showed no viability since day 15 after freezing. The spermatozoa frozen and stored at -6$0^{\circ}C$ or -8$0^{\circ}C$ showed higher post-thaw motility and viability on day 2, 1, 15 and 30 after freezing than that frozen and stored at-2$0^{\circ}C$(p<0.01), with no difference between two groups. Among different composition groups of the semen diluents of control(tris + egg yolk + glycerol), egg yolk-free, 히ycerol-free, and tris-free, Prior to freezing, the egg yolk-free diluent showed significantly love. motility and viability than the other diluents(p<0.05). On each thawing day (from day 2 to 15 after freezing), control group showed considerably higher motility and viability than the other groups(p<0.01). The canine spermatozoa frozen and stored at -6$0^{\circ}C$ and -8$0^{\circ}C$ showed gradual decrease of motility from day 2 to 30 after freezing and the spermatozoa of these two groups thawed on day 30 showed considerably love. motility than those thawed on day 2 after freezing, respectively(p<0.01). These results indicate that the freezing temperature of either -6$0^{\circ}C$ or -8$0^{\circ}C$ can be applicable to the freezing method using methanol and also all of the components of the semen diluent including cryoprotectant, buffer and cold-shock buffer are very important to maintain motility and viability of canine spermatozoa in the freezing and thawing procedure.

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농작물 조직배양의 육종학적 이용에 관한 연구 제1보 벼 약배양효율에 대한 유전자형과 다른 요인간의 복합효과 (Studies on the Application of Tissue Culture for Plant Breeding I. Effect of genotype and other combined factors on the callus induction and plant regeneration in anther culture of rice)

  • 김광호
    • 한국작물학회지
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    • 제28권4호
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    • pp.425-430
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    • 1983
  • 벼의 약배양효율을 증진시키는데 관여하는 요인들의 중요성을 검토하기 위하여 13개의 품종, 4개조합의 F$_1$, 7개조합의 F$_2$를 공시하여 이들의 약을 R$_2$배지에서 배양하였다. 유전자형간 약배양능력의 차이와 함께 약에 대한 저온처리, 배지의 당함량, 배지에 대한 생장조절제 첨가 등의 단독효과 및 유전자형과의 복합효과가 검토될 수 있도록 실험이 수행되었다. 1. 동일한 환경에서 재배된 벼품종 및 교배조합별 F$_1$들의 약배양능력간에는 큰 차이가 인정되었으며 포장에서 재배된 F$_2$들도 교배조합간에 평균 약배양능력이 크게 달랐다. 2. 동일한 교배조합의 F$_2$개체들 간에도 Callus분화율이 큰 변이를 보이고 있어 약배양능력이 유전변이를 보이는 형질임이 확인되었다. 3. 약을 배지에 치상하기 전 8$^{\circ}C$에 5-10일간 온도처리하는 것은 Callus 분화율을 향상시키는데 효과적이었는데 그 효과는 약배양능력이 높은 품종에서 더 컸다. 4. R$_2$배지의 당함량은 3-4%인 것이 Callus분화에 좋았으며 3-5%사이에서는 유전자형과의 복합효과가 인정되지 않았다. 5. R$_2$배지에서는 NAA만을 첨가하는 것이 Callus 분화율을 높이는데 효과적이었으며 약배양능력이 높은 유전자형을 이용했을 때 그 효과가 훨씬 더 컸다. 6. 약배양능력이 높은 유전자형을 선발, 이용하는 것이 약배양에 관여하는 요인들의 효율도 높일 수 있어 그 중요성이 가장 크다고 할 수 있다.

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Stellite bearings for liquid Zn-/Al-Systems with advanced chemical and physical properties by Mechanical Alloying and Standard-PM-Route

  • Zoz, H.;Benz, H.U.;Huettebraeucker, K.;Furken, L.;Ren, H.;Reichardt, R.
    • 한국분말야금학회:학술대회논문집
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    • 한국분말야금학회 2000년도 춘계학술강연 및 발표대회 강연 및 발표논문 초록집
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    • pp.9-10
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    • 2000
  • An important business-field of world-wide steel-industry is the coating of thin metal-sheets with zinc, zinc-aluminum and aluminum based materials. These products mostly go into automotive industry. in particular for the car-body. into building and construction industry as well as household appliances. Due to mass-production, the processing is done in large continuously operating plants where the mostly cold-rolled metal-strip as the substrate is handled in coils up to 40 tons unwind before and rolled up again after passing the processing plant which includes cleaning, annealing, hot-dip galvanizing / aluminizing and chemical treatment. In the liquid Zn, Zn-AI, AI-Zn and AI-Si bathes a combined action of corrosion and wear under high temperature and high stress onto the transfer components (rolls) accounts for major economic losses. Most critical here are the bearing systems of these rolls operating in the liquid system. Rolls in liquid system can not be avoided as they are needed to transfer the steel-strip into and out of the crucible. Since several years, ceramic roller bearings are tested here [1.2], however, in particular due to uncontrollable Slag-impurities within the hot bath [3], slide bearings are still expected to be of a higher potential [4]. The today's state of the art is the application of slide bearings based on Stellite\ulcorneragainst Stellite which is in general a 50-60 wt% Co-matrix with incorporated Cr- and W-carbides and other composites. Indeed Stellite is used as the bearing-material as of it's chemical properties (does not go into solution), the physical properties in particular with poor lubricating properties are not satisfying at all. To increase the Sliding behavior in the bearing system, about 0.15-0.2 wt% of lead has been added into the hot-bath in the past. Due to environmental regulations. this had to be reduced dramatically_ This together with the heavily increasing production rates expressed by increased velocity of the substrate-steel-band up to 200 m/min and increased tractate power up to 10 tons in modern plants. leads to life times of the bearings of a few up to several days only. To improve this situation. the Mechanical Alloying (MA) TeChnique [5.6.7.8] is used to prOduce advanced Stellite-based bearing materials. A lubricating phase is introduced into Stellite-powder-material by MA, the composite-powder-particles are coated by High Energy Milling (HEM) in order to produce bearing-bushes of approximately 12 kg by Sintering, Liquid Phase Sintering (LPS) and Hot Isostatic Pressing (HIP). The chemical and physical behavior of samples as well as the bearing systems in the hot galvanizing / aluminizing plant are discussed. DependenCies like lubricant material and composite, LPS-binder and composite, particle shape and PM-route with respect to achievable density. (temperature--) shock-reSistibility and corrosive-wear behavior will be described. The materials are characterized by particle size analysis (laser diffraction), scanning electron microscopy and X-ray diffraction. corrosive-wear behavior is determined using a special cylinder-in-bush apparatus (CIBA) as well as field-test in real production condition. Part I of this work describes the initial testing phase where different sample materials are produced, characterized, consolidated and tested in the CIBA under a common AI-Zn-system. The results are discussed and the material-system for the large components to be produced for the field test in real production condition is decided. Outlook: Part II of this work will describe the field test in a hot-dip-galvanizing/aluminizing plant of the mechanically alloyed bearing bushes under aluminum-rich liquid metal. Alter testing, the bushes will be characterized and obtained results with respect to wear. expected lifetime, surface roughness and infiltration will be discussed. Part III of this project will describe a second initial testing phase where the won results of part 1+11 will be transferred to the AI-Si system. Part IV of this project will describe the field test in a hot-dip-aluminizing plant of the mechanically alloyed bearing bushes under aluminum liquid metal. After testing. the bushes will be characterized and obtained results with respect to wear. expected lifetime, surface roughness and infiltration will be discussed.

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