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Study on Development of Canine Oocytes Treated by In Vitro Fertilization and ICSI

  • Park, Ji-Hoon;Chung, Young-Ho;Kim, Sang-Keun
    • Reproductive and Developmental Biology
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    • v.35 no.2
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    • pp.137-141
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    • 2011
  • These study was to investigate the in vitro fertilization and viability of fresh and vitrified oocytes. Also, the developmental capacity of IVF and intracytoplasmic sperm injection (ICSI) oocytes were investigated. Then vitrification was performed with the use of 20% ethylene glycol + 20% DMSO + 0.5 M sucrose + 10% FCS + TCM-199 medium. Vitrification immature oocytes are cultured in vitrification solution for 10 min afterwards transferred to expose at room temperature for 5 min. and transferred to the ice water for 5 min. The oocytes were sealed in a 1.0 mm straw and placed in a $LN_2$ container. Frozen oocytes were rapidly thawed in a water bath at $30{\sim}35^{\circ}C$, and then placed in TCM-199 medium containing 0.5 M sucrose for 5 min each, respectively, at $38^{\circ}C$. After being washed for 2~3 times, using fresh medium the oocytes were cultured in TCM-l99 medium supplemented with 5% FCS at $38^{\circ}C$ in 5% $CO_2$ and air. The normal morphology of fresh and vitrified-thawed oocytes were $87.1{\pm}2.1%$ and $54.8{\pm}2.5%$, respectively. The viability rates of fresh and vitrified-thawed oocytes were $70.0{\pm}2.2%$ and $41.9{\pm}2.6%$, respectively. Viability rates of vitrified-thawed oocytes were lower than that of fresh follicular oocytes (p<0.05). The in vitro maturation rates of fresh and vitrified oocytes were $45.1{\pm}3.6%$ and $28.9{\pm}4.4%$, respectively. The IVF rates of fresh follicular and vitrified-thawed oocytes were 34.00.2% and $20.2{\pm}2.6%$, respectively. The in vitro maturation and fertilization rates of vitrified-thawed oocytes were lower than those of the fresh follicular oocytes (p<0.05). A total of 350 oocytes were fixed and stained after co-incubation with spermatozoa, of which 88 had identifiable nuclear material. After IVF for 20 hrs, $25.1{\pm}3.4%$ of the oocytes found to have been penetrated by spermatozoas. Oocytes were fixed and stained after ICSI, and 105 oocytes contained identifiable nuclear material. After IVF and ICSI for 20 hrs, $34.3{\pm}3.4%$ and $59.0{\pm}2.0%$ of the oocytes were found to have been penetrated by spermatozoas. The developmental rates upon ICSI were significantly higher than those of the IVF method (p<0.05).

Combination Dyeing of Triacetate/PET Blended Fabric with Disperse Dye (트리아세테이트/PET 혼방 직물의 분산염료 혼합염색)

  • Kim, Myoung Ok;Lee, Jung-Soon
    • Science of Emotion and Sensibility
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    • v.19 no.4
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    • pp.3-12
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    • 2016
  • The aim of this study is to find the optimal combination dyeing condition for the enhancement of dye uptake and union dyeing of the composite material fabric made of triacetate and quick drying PET blended yarn. For the experiment, fabrics were one-bath combination dyed using the mixed dye of E-type disperse dye(C.I Disperse red 50) and S-type disperse dye(C.I. Disperse red 92) to measure dyed fabric's dye exhaustion, dye uptake, color and color difference according to the diverse conditions including dying temperature, time and mixed ratio of the dye. Dye equilibrium of combination dyeing occurred in $100^{\circ}C$, but by comparing dyed fabrics' K/S value and surface color, it was found that $120^{\circ}C$ was where the manifestation of color of triacetate and quick drying PET was identical. Mixed dye exhaustion and dye uptake merely changed as dyeing time increased, but color became more uniform. Therefore, it can be concluded that by using combination dyeing method, and by using the mixed dye which the mixing ratio of S-type dye and E-type dye is appropriately controlled, dye uptake can be improved compared to using single dyeing regardless of the color of E-type dye.

Determination of the effective components in the various parts of Luffa cylindrica (L.) Roemer and development to remove the flesh from its fruit : (II) -Removal of flesh on the net fiber In fruit of sponge-gourd and improvement of quality of the net fiber- (수세미외의 부위별(部位別) 유효성분(有效成分) 조사(調査) 및 사과락중(絲瓜絡中) 육질제거(肉質除去) 방법(方法) 개발(開發) 연구(硏究) : (II) -사과락 육질제거 및 섬유품질개선-)

  • Yoo, Tae-Bang;Chang, Ki-Woon;An, Byung-Chang;Shin, Jong-Sun;Park, Joung-Sang
    • Applied Biological Chemistry
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    • v.34 no.4
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    • pp.373-378
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    • 1991
  • The skin and fleshy substance on the net fiber of sponge-gourd fruit pressed mechanically was removed with 0.2% NaOH solution in $3{\sim}5$ hours. The treatment of 0.2% NaOH with 0.02% Monopol(non-ionogenic polyoxethylen derivative) as surfactnat and with 0.1% $Ca(OCl)_2$ as bleaching agent enhanced the effect to remove the fleshy substance and improve the quality of net fiber. Also, the wet hardness and tensile strength of net fiber were controlled by the crosslinkage of the fiber with glu tardialdehyde, glyoxal, and formalin, respectively. The net fiber was stable on the acid and alkaline solutions. Also the range of temperature to degradate the fiber was $338{\sim}385^{\circ}C$. These values indicated a fair stability. The improved net fiber can be used for raw material of bath, dish washing, oil and gas filter, and many kinds of decorations.

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Growth of Green Pepper(Capsicum annuum L.) in a Plastic Greenhouse Covered with Anti-dropping Plasma Film (방적성 Plasma 처리 필름으로 피복된 플라스틱온실의 풋고추 생육)

  • Chun, Hee;Kim, Kyung-Je;Kim, Jin-Young;Kim, Hyun-Hwan;Lee, Si-Young
    • Journal of Bio-Environment Control
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    • v.9 no.3
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    • pp.156-160
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    • 2000
  • The Plasma film treated with a high electric voltage was developed to enhance flow down of condensation drops on inside plastic film. Arch type greenhouse framed with iron pipe of 25mm diameter defand 1.5mm thickness were covered with either the developed plasma film or surfactant film(control). Green pepper seedlings raised for 40 days in plug trays were transplanted at a density of 110cm by 30cm in each greenhouse. The mount of condensational water on film surface, generated by 7$0^{\circ}C$ water bath chimney systems and flew down, was collected and measured. The amount of collected water after 150 minutes was 2.56 mL.100c $m^{-2}$ and 0.94mL.100c $m^{-2}$ , respectively, in the plasma film and surfactant film-covered greenhouses. The amount of condensational water drops attached on the cover at 08:20 a.m. at 60 days filter covering was 0.34mL.100c $m^{02}$ and 0.32mL.100c $m^{-2}$ , respectively, in the plasma film- and surfactant film-covered greenhouses. Solar irradiance transmitted into greenhouse was 2.0% higher in the greenhouse covered with the plasma film tan that in the greenhouse covered with the surfactant film. Air temperature in the plasma film-covered greenhouse was higher than the surfactant film-covered greenhouse by 0.5$^{\circ}C$. However, there was no difference in relative humidity between the two greenhouse. Plant height, leaf area, dry weight and early yield showed no significant differences.s.

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Study on Meat Tenderness of a Pretense Extracted from Domestic Pear (국산배에서 추출한 단백질 분해효소의 식육 연화제로서의 활용에 관한 연구)

  • Han Seung K.;Chin Koo B.
    • Food Science of Animal Resources
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    • v.24 no.4
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    • pp.326-328
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    • 2004
  • Domestic pear has been reported that it contained a pretense, which used for tenderizer of meat, however no researches for optimum level of the enzyme with maximum tenderness effect have been studied. Thus, this study was peformed to determine the optimum level of a protease for meat tenderness. Moisture contents (%) of domestic pears was determined. A pretense was homogenized in a mixer and centrifuged at 10,000 G for 1hr. After taken the supernatant, dialysis was conducted to remove salts and sugars, and freeze-dried. Then, various level (0.05, 0.1 and 0.2%) of the purified pretense were added to pork loins (3cm thickness). Then, pork samples were boiled at 80 for 12 min in a water bath to reach the interval temperature of 71 and chilled in an ice. Moisture contents (%) of domestic pears ranged from 87.2 and 87.8%. No differences in cooking loss of pork meats were observed (p>0.05) among various levels of a pretense. After centrifugation, the protein concentrations of a protease showed from 5.96 $\mu\textrm{g}$/fmL to 7.25 $\mu\textrm{g}$/mL. Increased level of a pretense up to 0.1% reduced (p<0.05) the shear value (kg/g), however no further reduction of shear value was observed at the level of higher than 0.1% of the purified pretense. The approximate molecular weight of the pretense analysed by sodium-dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) was 30 kDa. These results suggest that the optimum level of a pretense for the maximum effect of meat tenderness is above 0.1%. Further research will be peformed to determine the effect of various domestic pears and ingredients, such as salt and phosphate, on meat tenderness.

Preparation of in situ Patterned ZnO Thin Films by Microcontact Printing (Microcontact Printing을 이용한 미세패턴 ZnO 박막 제조)

  • 임예진;윤기현;오영제
    • Journal of the Korean Ceramic Society
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    • v.39 no.7
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    • pp.649-656
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    • 2002
  • In situ patterned zinc oxide thin films were prepared by precipitation of Zn(NO$_3$)$_2$ aqueous solution containing urea and by microcontact printing using Self-Assembled Monolayers(SAMs) on A1/SiO$_2$/Si substrates. The visible precipitation of Zn(OH)$_2$ that was formed in the Zn(NO$_3$)$_2$ aqueous solution containing urea was enhanced with an increase of the reaction temperature and the amount of urea. As the reaction time of Zn(NO$_3$)$_2$ with urea was prolonged, the thickness and grain size of Zn(OH)$_2$ thin layers were increased, respectively. The optimum precipitation condition was at 80$\^{C}$ for 1 h for the solution with the ratio of Zn(NO$_3$)$_2$ to urea of 1 : 8. Homogeneous ZnO thin films were fabricated by the heat treatment of 600$\^{C}$ for 1 h of Zn(OH)$_2$ precipitation on Al/SiO$_2$/Si substrate. This was available to the in-situ patterned ZnO thin films with uniform grain size. Hydrophobic SAM, Octadecylphosphonic Acid(OPA) and hydrophilic SAM, 2-Carboxyethylphosphonic Acid(CPA) were applied on the Al/SiO$_2$/Si substrate by microcontact printing method. In situ patterned ZnO thin film was successfully prepared by the heat treatment of Zn(OH)$_2$ precipitated on the surface of hydrophilic SAM, CPA.

Physiochemical Properties of Functional Oils Produced Using Red Yeast-Rice Ethanol Extracts and Diacylglycerol Oil (홍국쌀 에탄올 추출물과 Diacylglycerol Oil을 이용하여 제조한 기능성 유지의 이화학적 특성 연구)

  • Kim, Nam-Sook;Lee, Ki-Teak
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.36 no.2
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    • pp.201-208
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    • 2007
  • Functional oils (FOs) were produced from commercial diacylglycerol oil and red yeast rice extracts from 80% ethanol for 1 hr in a shaking water bath at $35^{\circ}C$ and 175 rpm. FOs contained (A) 600, (B) 1200, (C) 1800, and (D) 2280 ppm of red yeast-rice extracts, respectively. The Hunter a value and b value were risen whereas L value was reduced along with the increase of extract concentration. Content of monacolin K and total phenolic compounds in FOs significantly increased according to the increase of extract concentration. The oxidation stability of FOs was observed by Rancimat at $98^{\circ}C$. Induction time decreased according to the increase of extract concentration. The major volatile compounds of FOs were compared using the electronic nose (EN) system and solid phase microextraction (SPME) method combined with gas chromatograph/mass spectrometry (GC/MS). EN was composed of 12 different metal oxide sensors. Sensitivities (Rgas/Rair) of sensors from EN were analyzed by principal component analysis (PCA), whose proportion was 99.66%. For qualitative or quantitative analysis of volatile compounds by SPME-GC/MS, the divinylbenzene/carboxene/polydimethyl-siloxane fiber and sampling temperature of $50^{\circ}C$ were applied.

Preparation of nanoparticles CuInSe2 absorber layer by a non-vacuum process of low cost cryogenic milling (저가의 cryogenic milling 비진공법을 이용한 나노입자 CuInSe2 광흡수층 제조)

  • Kim, Ki-Hyun;Park, Byung-Ok
    • Journal of the Korean Crystal Growth and Crystal Technology
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    • v.23 no.2
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    • pp.108-113
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    • 2013
  • Chalcopyrite material $CuInSe_2$ (CIS) is known to be a very prominent absorber layer for high efficiency thin film solar cells. Current interest in the photovoltaic industry is to identify and develop more suitable materials and processes for the fabrication of efficient and cost-effective solar cells. Various processes have been being tried for making a low cost CIS absorber layer, this study obtained the CIS nanoparticles using commercial powder of 6 mm pieces for low cost CIS absorber layer by high frequency ball milling and cryogenic milling. And the CIS absorber layer was prepared by paste coating using milled-CIS nanoparticles in glove box under inert atmosphere. The chalcopyrite $CuInSe_2$ thin films were successfully made after selenization at the substrate temperature of $550^{\circ}C$ in 30 min, CIS solar cell of Al/ZnO/CdS/CIS/Mo structure prepared under various deposition process such as evaporation, sputtering and chemical vapor deposition respectively. Finally, we achieved CIS nanoparticles solar cell of electric efficient 1.74 % of Voc 29 mV, Jsc 35 $mA/cm^2$ FF 17.2 %. The CIS nanoparticles-based absorber layers were characterized by using EDS, XRD and HRSEM.

Rheological Properties of Gelatinized Model Foods (모형식품의 리올로지 특성)

  • Chun, Ki-Chul;Park, Young-Deok;Chang, Kyu-Seob
    • Korean Journal of Agricultural Science
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    • v.22 no.1
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    • pp.103-109
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    • 1995
  • The model foods were prepared by simulating mositure, protein and starch, and they were heated for 30 mins, at $80^{\circ}C$ and then cooled at $25^{\circ}C$ in water bath. Their rheological properties were investigated by the use of Brookfield wide-gap rotational viscometer at $30{\sim}60^{\circ}C$, and the rotation speed ranged from 0.6 to 6 rpm and solid content ranged from 8% to 11%, the results obtained were as follows. 1. All the model foods ($P_1S_3$, $P_2S_2$, $P_1S_1$, $P_2S_1$, $P_3S_1$, $P_4S_0$) exhibited pseudoplastic behaviors with yeild stress and were thixotropic foods which showed time - dependent structural decays, but the starch food of 8 ~ 11 % solid content did not show the flow behavior. 2. The correlation between the rheological parameters and the protein content of model foods in various moisture content did not appeared a constant relationship. 3. The change of shear stress against shear rate in high starch foods was larger than that in high protein foods and the structure at initial shear time was decayed with a quatic equation according to the Tiu's Model and structural decay was in parallel with the increase of shear rate. 4. The temperature dependency of the apparent viscosity of $P_1S_2$, and $P_2S_1$ was fully expressed by Arrhenius equation and activation energies of their food were 2.35 and $1.34Kcal/g{\cdot}mol$, respectively.

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Effect of Hypo-osmotic Swelling (HOS) Test on Subsequent Post-thaw Testicular Spermatozoa (고환조직 동결-융해 후 회수된 고환 정자에 대한 Hypo-osmotic Swelling (HOS) Test의 효과)

  • Park, Yong-Seog;Lee, Hyoung-Song;Song, Sang-Jin;Kim, Jeong-Wook;Kang, Inn-Soo;Seo, Ju-Tae
    • Clinical and Experimental Reproductive Medicine
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    • v.27 no.3
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    • pp.267-273
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    • 2000
  • Objectives: We have previous reported that thawed testicular sperm and sperm extracted from seminiferous tubule could achieved optimal fertilization and pregnancy in azoospermic patients. However, thawed testicular sperm did not show motility in many cases. Therefore we studied viability of immotile sperm extracted from frozen-thawed seminiferous tubule using hypo-osmotic swelling (HOS) test and eosin-Y test. Materials and Methods: After sperm extraction using for ICSI, the remained sections of seminiferous tubules were frozen with a computerized freezer. For thawing and preparation of testicular sperm, the seminiferous tubules were thawed by removing from $LN_2$ and letting them at room temperature for 10 min followed by %37^{\circ}C$ water bath for 10 min. The prepared samples were washed for free of preservation medium and sperm preparation method described previous. Sperm was suspended in 0.1 ml hypoosmotic solution. After 30 minutes, the type of distally coiled sperm were assessed. Results: In 44 cases of cryopreservation of seminiferous tubules in obstructive azoospennic patients, the fertilization rates with 2PN were 71.4% and pregnancy rates were 34.1%. The presence of motile spermatozoa on subsequent post-thaw testicular sperm remarked 15.1% and were increased to 77.3% just before ICSI. After sperm extracted from frozen-thawed seminiferous tubule, 3 hrs later in in vitro culture, the cases of presence of motile sperm, reaction of hypo-osmotic swelling test and viable sperm were 63.6% (28/44), 93.2% (41/44), and 77.3% (34/44), respectively. Conclusions: Just after post-thawed testicular sperm did not showed motility. Although motility was gained after in vitro culture, many cases showed non-motile sperm until optimal insemination time. However, HOS test showed positive reaction in non-motile sperm. Therefore, HOS test is an alternative method for the selection of viable sperm for ICSI.

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