• Title/Summary/Keyword: Pre-freezing

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Change of Sperm Viability and Acrosome Integrity of Post-thawed Korean Jeju Black Bull Spermatozoa according to Glycerol Concentration (제주 흑우 동결 정액 제조에 있어 Glycerol의 농도에 따른 생존율 및 정자 첨체 양상의 변화)

  • Choi, Sun-Ho;Ko, Min-Hee;Kang, Tae-Young;Cho, Sang-Rae;Park, Yong-Sang;Oh, Shin-Ae
    • Journal of Embryo Transfer
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    • v.26 no.3
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    • pp.187-193
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    • 2011
  • This study was carried out to establish most suitable freezing condition, to evaluate the different glycerol concentration of freezing and thawing rates on motility, viability, membrane integrity and acrosome intecrity of frozen Korean Jeju Black Bull spermatozoa, Semen was collected from a Korean Jeju Black Bull using an artificial vagina and transported to the laboratory. The semen was extended gradually 1:5 then cooled slowly for 2 hrs to 4$^{\circ}C$. The semen was diluted 1:1 with cryoprotectant extenders (3%, 5% and 7% glycerol) and equilibrated for 2 hrs at cold chamber and packed to 0.5 ml straws. The semen straws were located above 3 cm of liquid nitrogen for 5 minutes, above 5 cm for 10 min and above 8 cm for 10 min. And then the frozen straw was plunged into LN$_2$. The presented straws were examined the viability and motility after thawed at 37$^{\circ}C$ water bath. The viability and membrane integrity immediately post-thawing were significantly higher in samples frozen in 7% glycerol than 3% and 5% glycerol (p<0.05). After CTC staining to assess acrosome integrity, F pattern was significantly increased, but B pattern was significantly decreased in 7% glycerol (p<0.05). Freezing distance of 5 cm from liquid nitrogen and pre-cooling for 10 min yield better survival and membrane integrity, but not significant difference. However, AR pattern according to CTC staining was significantly decreased in 3 cm for 5 min.

Angelica gigas Nakai 현탁세포 배양의 동결보존 연구

  • Jo, Ji-Suk;Lee, Song-Jae;Jeon, Su-Hwan;Kim, Ik-Hwan;Kim, Dong-Il
    • 한국생물공학회:학술대회논문집
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    • 2000.04a
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    • pp.321-324
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    • 2000
  • Cell culture of Angelica gigas Nakai producing decursin and decursinol angelate as secondary metabolites were preserved in liquid nitrogen after pre-freezing in deep $freezer(-70^{\circ}C)$. The development of optimal procedure for cryopreservation was investigated by using cryoprotectant and pre-treatment before cooling. 0.7 M sucrose was found be the optimum osmotic pre-conditioning culture medium compared to mannitol, sorbitol and NaCl with the same osmotic pressure. In pre-culture medium, cells in exponential phase, supported the best growth after cryopreservation. Effective cryoprotectant was proved to be a mixture of sucrose, glycerol, DMSO. Higher concentration of cryoprotectant was better for cell viability. The highest relative cell viability established after the development of optimal procedure was 65%.

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The Effects of the Pre-treatments with Proteins on Dyeing of Silk Fabric with Caesalpinia sappan (견직물의 소목염색에서 단백질 전처리 효과)

  • Hwang, So Hee;Jang, Jeong Dae
    • Textile Coloration and Finishing
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    • v.28 no.3
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    • pp.208-218
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    • 2016
  • This study investigated the effects of the working mechanisms of proteins, mordants, and dyes, as well as the mordanting order, on dye uptake by silk fabric pre-treated with proteins and dyed with freeze-dried sappan wood water extract. Soybean protein and sodium caseinate were used as the proteins. 1. When Al mordants were not used, the dyeability of the fabrics increased upon protein pre-treatment as compared to the case without treatment. 2. Dyeing with protein pre-treatment, followed by mordanting, led to the highest dye uptake, and the optimal protein concentration was 5%. 3. The K/S values slightly decreased with an increase in the dyeing temperature, and the fabric turned dark red in color when dyeing was carried out at increasing temperature. Fabrics showed the highest dye uptake at $40^{\circ}C$. 4. Regarding the effect of time, the K/S values of the fabrics with and without protein treatment showed almost no increase after the initial dyeing time of 10min; further, there was hardly any difference in the cases with and without protein pre-treatment. 5. In case of protein pre-treatment fabrics, the washing fastness was level 2. The dry cleaning fastness showed very excellent result with level 4-5. The rubbing fastness was better in dry rubbing than in wet rubbing of the fabrics. For the light fastness, all dyed fabrics showed low fastness.

Effect of Kind of Sugar Added in Tris-buffer on Motility of Post-thaw Spermatozoa in Canine (Tris-buffer에 첨가되는 당의 종류가 동결ㆍ융해정자의 운동성에 미치는 영향)

  • 유대중;정수룡;오인석;김흥률;이계웅;조성균;배인휴;양철주;공일근
    • Journal of Embryo Transfer
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    • v.17 no.2
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    • pp.137-143
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    • 2002
  • This study was conducted to evaluate the effect of sugar kinds and combination of various sugars, and straw size and pre-freezing time on motility of post-thaw spermatozoa in canine. In general, the extender was supplemented with fructose or glucose for semen freezing. The extender used in .this study was Tris-citric acid extender (Tris-buffer) supplemented with 20% Egg-yolk, 8% glycerol, 1% Equex STM paste, and 70 mM sugars such as monosaccharide (fructose and xylose) and disaccharide(trehalose). To evaluate of sugar combination, the sugars supplemented in Tris-buffer. were combined such as control (fructose, xylose, trehalose), two combinations (Fru+Tre, Fru+Xy1, Tre+Xy1) and three combinations (Fru+Tre+Xy1). The motility after CASA analysis in Fru+Tre+Xy1 was higher than those in fructose, trehalose, xylose, Fru+Tre, Fru+Xy1, Tre+Xy1 (69 vs. 58, 61, 50, 65, 20, 54). However, the progressive motility after CASA analysis in Fru+Tre group was higher than those in fructose, trehalose, xylose, Fru+Xy1, Tre+Xy1, Fru+Tre+Xy1 (59% vs. 47, 55, 42, 13, 49, 44%). The survival rate of post-thaw spermatozoa in 0.25 $m\ell$ straw for 10 min pre-freezing was significantly higher than those in 0.25 $m\ell$ straw for 10 min, 0.25 and 0.5 $m\ell$ for 5 min (80+0.0 vs. 65+7, 68+16, 58+8%; P<0.05). The results indicated that the progressive motility of post-thaw spermatozoa in 70 mM Fru +Tre (two combination) following CASA analysis and 0.25 ml straw for 10 min pre-freezing time could be better for freezing of semen in canine.

A Trifloxystrobin Fungicide Induces Systemic Tolerance to Abiotic Stresses

  • Han, Song-Hee;Kang, Beom-Ryong;Lee, Jang-Hoon;Lee, Seung-Hwan;Kim, In-Seon;Kim, Chul-Hong;Kim, Young-Cheol
    • The Plant Pathology Journal
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    • v.28 no.1
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    • pp.101-106
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    • 2012
  • Trifloxystrobin is a strobilurin fungicide, which possesses broad spectrum control against fungal plant diseases. We demonstrated that pre-treating red pepper plants with trifloxystrobin resulted in increased plant growth and leaf chlorophyll content compared with those in control plants. Relative water content of the leaves and the survival rate of intact plants indicated that plants acquired systemic tolerance to drought stress following trifloxystrobin pre-treatment. The recovery rate by rehydration in the drought treated plant was better in those pre-treated with trifloxystrobin than that in water treated plants. Induced drought tolerance activity by trifloxystrobin was sustained for 25 days after initial application. The trifloxystrobin treated red pepper plants also had induced systemic tolerance to other abiotic stresses, such as frost, cold, and high temperature stresses. These findings suggest that applying the chemical fungicide trifloxystrobin induced systemic tolerance to certain abiotic stresses in red pepper plants.

The Evaluation of Various Conditions in the Cryopreservation of Mouse Embryos - Rapid and Slow Method of Cryopreservation, Culture Media and Cell Stages (생쥐배아의 냉동보존에 있어서 여러 조건의 평가 - 저속 처리단계와 급속 처리단계, 배양액, 세포기)

  • Yi, Seung-Yeun;Kwon, Ju-Taek;Song, Hee-Won;Cho, Yun-Hee;Lee, Ky-Sook;Rheu, Cheul-Hee;Kim, Jong-Duk
    • Clinical and Experimental Reproductive Medicine
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    • v.26 no.2
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    • pp.127-135
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    • 1999
  • Cryopreservation is able to store the surplus pre-embryos for freezing and furthermore thawing and transfer in a subsequent cycle. Cryopreserving cells which are maintaining their viability are the very complex process. This study has been carried out in order to find the effects of cryopreservation steps, freezing media and embryonic stages on the rates of viability and development of cryopreserved mouse embryos. Female ICR mice ($6{\sim}8$ weeks old) were induced to superovulate by sequential intraperitoneal injection of 5 IU PMSG and 5 IU hCG 48h apart. Mouse embryos were collected according to its developmental stage after the injection of hCG. Embryos were cryopreserved not only by cryoprotectant step (1 step${\sim}$4 step) but also in a variety of media (HTF, IVF medium, D-PBS) and cell stage. The results were as follows: There is no clear advantage in these freezing media of rapid method, but 4 cell and 8 cell of slow method (2, 3, 4 step) have advantage in D-PBS. The development of embryos according to cell stage become greater in 8 cell stage. In the treatment steps of cryopreservation, the development of embryo to blastocyst was similar among rapid method, but the development of 4 cell and 8 cell embryos to blastocyst according to slow method was better than rapid method.

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An Experimental Study for the Effective Use Scheme of Snow Removal Materials on Road (도로 제설재의 효과적 사용방안에 관한 실험적 연구)

  • Do, Jongnam;Kim, Taesoo;Lee, Chanbok;Kim, Yeonjoong;Chun, Byungsik
    • Journal of the Korean GEO-environmental Society
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    • v.13 no.2
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    • pp.5-17
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    • 2012
  • The amount of snow due to global warming and climate change has recently increased. The effective management of snow removal materials will be required. First, on the basis of domestic winter temperature, it is not necessary to get a baseline to less than the freezing point of $-52^{\circ}C$ for the calcium chloride($CaCl_2$) of 30%.. In terms of cost comparison between calcium chloride($CaCl_2$) and sodium chloride($NaCl$), the calcium chloride($CaCl_2$) is 2 to 3 times more expensive, and the supply of the calcium chloride($CaCl_2$) is not produced in domestic country and is in the conditions that have to imports all needed. Accordingly, the effective use scheme of snow removal materials should be considered to multifaceted ways. Thus, the objective of this study is to develop effective method and to replace from calcium choride($CaCl_2$) to sodium chloride($NaCl$) solution in the current snow removal operating system that uses a pre-wetted salt spreading method. The effective method that equals to the quality of the existing snow removal materials was developed in this study through performance tests for deicing chemicals, corrosion test of steel and freezing and thawing tests of concrete.

Evaluation on Performance of Repair Mortar Used for Pre-wetting Spray Method (프리웨팅 스프레이 공법용 모르타르의 성능평가)

  • Nam, Yong-Hyuk;Chung, Young-Jun;Jang, Suk-Hwan;An, Young-Ki;Kim, Sung Chil
    • Journal of the Korea institute for structural maintenance and inspection
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    • v.9 no.4
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    • pp.235-242
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    • 2005
  • This study is on the evaluation of performance of polymer cement mortar which is used for pre-wetting spray method. Pre-wetting spray method is an epoch-making method to repair concrete structures damaged, which is added a small quantity water preciously to dry mortar to reduce dust and rebound and spray mortar mixed with fixed quantity water at nozzle before spray. The result showed that physical performance such like compressive, flexural and adhesive strength of polymer cement mortar, TS 100 used for pre-wetting spray method was superior to other repair mortar. Also durable performance such as resistance on permeability of chloride ion, carbonation, chemical and freezing-thawing was excellent.

Seminal Plasma Heparin Binding Proteins Improve Semen Quality by Reducing Oxidative Stress during Cryopreservation of Cattle Bull Semen

  • Patel, Maulikkumar;Gandotra, Vinod K.;Cheema, Ranjna S.;Bansal, Amrit K.;Kumar, Ajeet
    • Asian-Australasian Journal of Animal Sciences
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    • v.29 no.9
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    • pp.1247-1255
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    • 2016
  • Heparin binding proteins (HBPs) are produced by accessory glands. These are secreted into the seminal fluid, bind to the spermatozoa at the time of ejaculation, favour capacitation, acrosome reaction, and alter the immune system response toward the sperm. The present study was conducted with an objective to assess the effect of purified seminal plasma-HBPs (SP-HBPs) on cross bred cattle bull sperm attributes during two phases of cryopreservation: Pre freezing and freezing-thawing. SP-HBPs were purified from pooled seminal plasma by heparin affinity chromatography. Three doses of SP-HBPs i.e. 10, 20, $40{\mu}g/mLs$ semen were standardized to find out the optimum dose and $20{\mu}g/mLs$ was found to be an optimum dose. Semen as such and treated with SP-HBPs was diluted with sodium citrate-egg yolk diluter and cryopreserved as per the standard protocol. Sperm parameters i.e. motility, viability, Hypo-osmotic swelling test (HOST), acrosome damage, in vitro capacitation and lipid peroxidation were evaluated in SP-HBP treated and untreated (control) semen at both phases of cryopreservation. A considerable variation in percent sperm motility, viability, membrane integrity (HOST), acrosome damage, acrosome reaction and lipid peroxidation was observed at both phases among the bulls irrespective of the treatment. Incubation of neat semen with $20{\mu}g/mL$ SP-HBP before processing for cryopreservation enhanced the average motility, viability, membrane integrity by 7.2%, 1.5%, 7.9%, and 5.6%, 6.6%, 7.4% in pre-frozen and frozen-thawed semen in comparison to control. There was also an average increase of 4.1%/3.9% in in vitro capacitation and acrosome reaction in SP-HBPs-treated frozen-thawed semen as compared to control. However, binding of SP-HBPs to the sperm declined acrosome damage and lipid peroxidation by 1.3%/4.1% and 22.1/$32.7{\mu}M$/$10^9$ spermatozoa in SP-HBP treated pre-frozen/frozen-thawed semen as compared to control, respectively. Significant (p<0.05) effects were observed only in motility, HOST and in vitro acrosome reaction. It can be concluded that treatment of neat semen with SP-HBPs before cryopreservation minimized the cryoinjury by decreasing the generation of reactive oxygen species.

Characteristics of Mortar at Low Temperature with De-icing Agency (시판 방동제의 저온 양생된 모르타르 특성)

  • 유성원;서정인
    • Proceedings of the Korea Concrete Institute Conference
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    • 2002.10a
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    • pp.33-38
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    • 2002
  • Concrete placed under cold weather has some defects such as the decrease of initial strength by hydration delay, strength unrecovery at unhardened concrete freezing, and structural failure and crack by expansion pressure. So, in this study, we tried to evaluate the JIS mortar which was made under cold weather using de-icing agency. In mortar test, the do-icing agency increased compressive strength under standard curing, and the de-icing agency made by NaNO$_2$ gave the highest strength. However, as pre-curing time under 21$^{\circ}C$ was short, the de-icing agency made by NaNO$_2$ and Ca[NO$_3$]$_2$ had the highest strength.

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