• Title/Summary/Keyword: extender

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Effects of Taurine and $\alpha$-Tocopherol Treatment during freezing on Sperm Characteristics and Function in Frozen-Thawed Porcine Semen (돼지 정액의 동결시 Taurine과 $\alpha$-Tocopherol 첨가가 동결$\cdot$융해 정자의 성상과 기능에 미치는 영향)

  • Shin H. A.;Kim C. K.;Chung Y. C.;Pang M. G.
    • Reproductive and Developmental Biology
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    • v.29 no.3
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    • pp.155-162
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    • 2005
  • The present study evaluated whether an exogenous antioxidants, taurine and $\alpha$-tocopherol, could, when added to the freezing extender, improve the post-thaw sperm characteristics, function, the level of reactive oxygen species (ROS) generation, and the level of lipid peroxidation (LPO) in frozen-thawed porcine semen. CASA (computer-aided sperm analysis), HOST (hypoos-motic swelling test), chemiluminescence using luminol and lucigenin and the detection of malondialdehyde for LPO was performed in frozen-thawed porcine sper-matozoa. The results obtained in these studies are as follows. While no beneficial effects of taurine and $\alpha$-tocopherol supplementation were visible in motility, viability, acrosome reaction, tail swelling patterns, and the generation of $O^{2-}$ of frozen-thawed porcine sper-matozoa, $H_{2}O_{2}$ was decreased by all treatments except taurine 50mM treatment. In conclusion the taurine and $\alpha$-tocopherol treatments during freezing reduced generation of reactive oxygen species and production of malondialdehyde in frozen-thawed porcine semen, and the ROS savangers may minimize various damages of spermatozoa during freezing.

Analysis of Frozen-Thawed Sperm Characteristic in Alpha 1,3-Galactosyltransferase(GalT) Knock-out Cloned Miniature Pig (Alpha 1,3-Galactosyltransferase (GalT) Knock-out 복제 미니돼지 정액의 동결-융해 후 정액 성상 분석)

  • Woo, Jea-Seok;Lee, Yong-Seung;Yoo, Han-Jun;Hwang, Seong-Soo;Oh, Keon-Bong;Cheong, Hee-Tae;Yang, Boo-Keun;Park, Soo-Bong;Park, Choon-Keun
    • Reproductive and Developmental Biology
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    • v.35 no.3
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    • pp.385-390
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    • 2011
  • The purpose of this study was undertaken to evaluate of cryopreservation efficiency in ${\alpha}$ 1,3-galactosyltransferase knock-out(GalT KO) cloned miniature pig sperm. To compare ability of frozen-thawed sperm characteristics, three different pig strains (GalT KO) cloned miniature pig, PWG miniature pig and Duroc were used. The ejaculated semen from the three pig species was diluted with same volume extender and added to LEY solution for freezing. The diluted semen was placed in 0.5 ml straws, and freezing was initiated by exposing the straws to liquid nitrogen ($LN_2$) vapours for 10 min before placing them into $LN_2$ for cryopreservation. After thawing, the sperm ability were assessed for viability (SYBR-14/PI staining), abnormality (Rose Bengal staining), and acrosome status (intactness, intensity and capacitation) (chlorotetracycline, CTC staining). The viability of frozen-thawed GalT KO pig sperm had no significant difference as compared with Duroc and PWG miniature pig sperm. However, The CTC pattern of frozen-thawed GalT KO cloned miniature pig spermatozoa showed significantly lower rates in F pattern and AR pattern (p<0.05) and significantly higher rates in B pattern than Duroc and PWG miniature pig (p<0.05). The abnormality of GalT KO cloned miniature pig sperm was significantly lower as compared to Duroc and PWG miniature pig sperm (p<0.05). In conclusion, GalT KO cloned miniature pig semen can be cryopreserved successfully and used for artificial insemination reasonably.

Evaluation of Glucose-BSA as Extender of Chilled Canine Spermatozoa for Long-term Storage (장기간 냉각 보존을 위한 개 정자 희석액으로서 Glucose-BSA의 유효성 평가)

  • Yu, I.J.
    • Journal of Embryo Transfer
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    • v.21 no.4
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    • pp.323-330
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    • 2006
  • 본 연구의 목적은 3주 동안 장기간 냉각상태로 보관된 개 정자의 기능적 특성을 알아보고자 두 종류의 정자 희석액, Glucose-BSA(G-BSA), Dimitropoulos-II(BIMI)가 정자의 기능적 특성에 미치는 영향을 알아보고자 한다. 개 정액을 수지법에 의해 채취하여 원심분리한 후 정장은 제거하였다. 정자를 G-BSA나 DIMI으로 희석하여 최종 농도를 $100\times10^6$ sperm/ml로 하였다. 희석된 정자를 정자 운송 시스템을 이용하여 루지애나 주립 대학에서 뉴올리언즈 실험실까지 운송하였다. 정자 운송 시스템은 $20^{\circ}C$에서 $9^{\circ}C$까지 분당 $0.08^{\circ}C$ 냉각율로 설정하였다. 희석된 정자는 $4^{\circ}C$에서 3주간 보관하였다. 보관 1일부터, 매주 단위로 정자의 활력, 정자 원형질막 고유성(생존성), 정자 첨체의 고유성을 검사하였다. 또한 체외조건하에서의 정자 번식 능력을 평가하기 위해 zona binding assay를 실시하였다. 실험 결과 냉각 상태로 3주 동안 보관된 개 정자는 여전히 정자의 기능적 특성을 나타냈다. 냉각기간이 길어질수록 정자의 활력 및 생존성은 감소하였으며(P<0.01), 첨체의 고유성은 냉각기간에 따라 감소하였으나 유의성은 없었다. 정자의 특성 중 활력이 다른 특성에 비해 냉각처리에 민감한 반응을 보였다. G-BSA배지에 보관된 정자의 활력 및 생존성이 DIMI에 비해 높게 나타났으며, 1 주 동안 보관된 정자의 생존성의 비교시 유의성이 높게 나타났다(P<0.05). 그러나 첨체의 고유성은 1일 동안 DIMI에 보관된 정자에서 높게 나타났다(P<0.05). 3주 동안 냉각상태로 보관된 개 정자는 여전히 난자의 투명대에 결합하는 능력을 보였으며, G-BSA에 보관된 정자의 경우 난자에 부착되는 평균 정자 수가 보관일수에 따라 유의적으로 감소하였다(P<0.05). 그러나 희석액 종류에 따른 난자에 부착되는 평균 정자 수의 유의적 차이는 없었다. 정자의 활력 및 생존성과 난자의 투명대에 결합하는 정자 수와의 상관관계를 본 결과, 활력 및 생존성이 높은 정자일수록 난자의 투명대에 많이 결합하여 정자의 잠재적 번식력이 증가함을 보였다. 결론적으로 G-BSA나 DIMI의 희석액으로 희석된 후 $4^{\circ}C$에서 냉각 보관된 정자는 3주 동안 정자의 기능적 특성을 유지하였으며 냉각 보관 2주까지 좋은 성적의 정자 특성을 나타냈다. 개 정자 냉각 보존에 대안적인 희석액으로 G-BSA도 사용 가능한 것으로 사료된다.

Investigation of the Korean Traditional Hobun(Oyster shell W.) Manufacturing Technique : Centering on Calcination Method (전통 호분(합분) 제조기술 연구 : 소성방법을 중심으로)

  • Lee, Han-Hyoung;Kim, Soon-Kwan;Kim, Ho-Jeong;Jeong, Hye-Young
    • Journal of Conservation Science
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    • v.23
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    • pp.103-118
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    • 2008
  • Hobun(Oyster shell W.) is a traditional material used as extender and white pigment from ancient times. The production method of it, however, has been discontinued. We have studied the traditional production method of Hobun through calcination of oyster shell, which is one of the traditional ways for preparing Hobun. Our work has the important meaning in that we can reproduce the manufacturing method of the discontinued traditional material and also it provides a solid background knowledge to stabilize the production and supply of Hobun for the cultural asset repairing materials. The result can be summarized as followings: The production processes of Hobun by calcination method are divided into 4 steps - calcination ${\rightarrow}$ slaking(pulverization) ${\rightarrow}$ separating fine powder by submergence in water ${\rightarrow}$ drying. In calcination step, the temperature is required to exceed $700^{\circ}$ to get pure white color of Hobun, since organic materials in the shell cause the final powder to be less white below $600^{\circ}$. And the calcination methods produce significant amount of calcium hydroxide, which is incongruent for pigment materials without additional treatments. The experimental study also demonstrated that the additional treatment process introduced in traditional paintings can be a probable process since the calcium of potassium hydroxide is observed to be promoted by this treatment. It is also concluded that, the calcination method of Hobun is appropriate for a small amount and high quality production.

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Comparison Study on the Material Characteristics of Oil Paints (I) (유화물감의 재질적 특성 비교 연구 (I))

  • Kim, Jung Heum;Park, Hye Sun;Lim, Sung Jin
    • Journal of Conservation Science
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    • v.33 no.2
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    • pp.85-95
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    • 2017
  • Oil paints are mixtures of pigments, drying oils and additives. In the past, oil paints were mainly composed of inorganic pigments. However, recently color matching techniques vary depending on manufacturers due to the development of various kinds of synthetic pigments. Despite this, most studies of oil paints in South Korea are about durability tests, and there is no comparative study on the characteristics of commercial oil paint components. This study aims to compare the properties of four different kinds of oil paints from four manufacturers, which are the most popularity used. Extender pigments in oil paint from C brand differed from that of other manufacturers and various kinds of coloring pigments were differently used depending on the oil paints and the manufacturers. The mixing ratios and the pigment types differed even for oil paints having the same product name. It is assumed that these differences could affect the colors. The result of this study is expected to contribute to the analysis of artworks through the accumulation of scientific data of oil paints. In addition, it can be utilized as a scientific basis for art history studies, including the characteristics of artists or production year.

Optimum Chain Extension and Change of the Average Particle Size of Aqueous Polyurethane Dispersion (수분산 폴리우레탄 제조시 최적 사슬연장 조건과 입도 변화에 관한 연구)

  • Kong, H.C.;Jhon, Y.K.;Cheong, I.W.;Kim, J.H.
    • Journal of Adhesion and Interface
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    • v.3 no.2
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    • pp.9-16
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    • 2002
  • In the synthesis of water-based polyurethane using self-emulsification process, after being neutralized, polyurethane pre-polymers containing ionic pendant groups are dispersed by simple convective mixing. Preparation of dispersion is followed by chain extension reaction, which is considered as important step for growth of the molecular weight of polyurethane. In this work, pH variations in the aqueous phase were monitored during the chain extension process in the presence of water-soluble diamines. The optimum degree of chain extension and the average particle size in the polyurethane dispersions were examined with varying ionic pendent group contents, type of chain extenders, and feed rate of chain extenders, The initial pH value in the aqueous phase linearly increased and the optimum chain extension point could be obtained from the intersection of two linear lines having different slopes, All average particle sizes before chain extension reaction were almost same, however, the final average particle size increased as feed rates of chain extenders increased, In addition, as the ionic pendant group contents increased, the particle size decreased since the hydrophilicity and hydrodynamic volume increased. As carbon numbers of the chain extenders increased, the final particle size increased significantly. From the results, it was concluded that the chain extension reaction took place among the particles not only in a particle.

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The Reduction of Hydrogen Peroxide in Viable Boar Sperm Cryopreserved in the Presence of Catalase (Catalase 첨가에 따른 돼지 정액 동결 및 융해 후 생존 정자에서 Hydrogen Peroxide의 감소)

  • Kim, Su-Hee;Lee, Young-Jun;Kang, Tae-Woon;Kim, Yong-Jun
    • Journal of Veterinary Clinics
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    • v.28 no.1
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    • pp.13-19
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    • 2011
  • Semen cryopreservation induces the formation of reactive oxygen species (ROS), and the ROS cause sperm damage. We aimed to investigate the effects of the antioxidative enzyme catalase (CAT) on sperm quality and ROS during cryopreservation. Sperm rich fractions collected from five Duroc boars were cryopreserved in freezing extender with (200 or 400 U/mL) or without CAT (control). After thawing, sperm motility, viability, normal morphology, plasma membrane integrity, mitochondrial function and intracellular ROS were evaluated. CAT significantly improved total sperm motility at a concentration of 400 U/mL (P < 0.05), but didn't improve progressive sperm motility, viability, morphological defects, plasma membrane integrity and mitochondrial function in frozen-thawed boar sperm. In evaluation of ROS, CAT had no effect on reduction in ${\cdot}O_2$, but scavenged $H_2O_2$ in viable frozen-thawed boar sperm at concentrations of 200 and 400 U/mL (P < 0.05). In conclusion, CAT was not enough to improve quality of frozen-thawed sperm, but can reduce $H_2O_2$ generation in viable boar sperm during cryopreservation.

The Effect of Ionic Group and MMA Contents on the Physical Properties of PU/PMMA Hybrids (PU/PMMA Hybrids의 물성에 대한 이온성기와 MMA함량의 영향)

  • Jeong, Chang Nam;Cho, Hang Kyu;Noh, Si Tae
    • Applied Chemistry for Engineering
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    • v.8 no.4
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    • pp.575-581
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    • 1997
  • UDs were synthesized from two different polyols(PTMG, PBEAG), ionic chain extender(DMPA), EDA with $H_{12}-MDI$. PU/PMMA hybrids were prepared with free radical polymerization of MMA monomer in MMA-swelled PUD. PUD particle size and film properties were investigated ionic content and polyol type. Mechanical and thermal properties of PU/PMMA hybrid film were studied in terms of PU's ionic content and the venation of PU/PMMA compositions. As DMPA content increased from 2wt% to 10wt% in PUD, particle size of PUD decreased. PUD's particle size with ester type polyol was found to be smaller then ether type polyol used. Phase separation between hard segment(HS) and soft segment(SS) with ionic contents in PU was shown by the thermal, mechanical property measurement. Although the composition of MMA was changed from 0 to 40 wt% in PU/PMMA hybrid, the particle size of the hybrid did not increase. Using the ester type polyol, tensile strength of hybrid was found to increase by 2wt% - 6wt% DPMA content, but as higher content the strength of hybrid decreased. Moreover with the ether type polyol, tensile strength of hybrid was observed to increase by 2wt% - 4wt% DMPA content, while decreasing at higher content. PU and PMMA polymer molecule being mixed in molecular level was confirmed from the pattern of $T_g$ in DSC thermogram.

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The Effects of Supplementation of BSA or Fatty Acid Free BAS on the Motility of Fresh or Cryopreserved Rooster Spermatozoa (BSA 또는 Fatty Acid Free BSA 첨가가 닭 희석 정자와 동결 정자의 생존성에 미치는 영향)

  • Kim, Sung Woo;Kim, Min Soo;Yu, Yeonhui;Kim, Chan-Lan;Jeon, Ik Soo;Kim, Chongdae
    • Korean Journal of Poultry Science
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    • v.44 no.1
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    • pp.59-65
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    • 2017
  • This study examined factors affecting the analysis of motility of chicken semen. The viability of spermatozoa was estimated using varying dilution ratios and supplementation with BSA or fatty acid free (FAF)-BSA as protein sources in semen diluent. Fresh semen was examined after preparing dilutions in beltsvile poultry semen extender (BPSE) of 1/8, 1/16 and 1/32 at $25^{\circ}C$. The motility of incubated semen at each dilution was observed at 3 min (89.9%, 69.9% and 53.2%), 30 min (86.7%, 71.4% and 51.7%), 1 h (89.5%, 74.0% and 53.5%) and 3 h (78.5%, 66.5% and 45.7%), respectively. The addition of BSA or FAF-BSA to BPSE diluent significantly increased the viability of semen in 1/32 dilution with results of 53.2% (control), 84.8% (BSA) and 92.9% (FAF-BSA) (p<0.05). This phenomenon was also observed in the dilution of frozen semen, where FAF-BSA treatment increased the viability of thawed semen from 17.6% to 34.0% in a 1/8 dilution (p<0.05). When the protein sources were used in the dilution, the survival rates of diluted chicken semen were also increased with time lapse. These results show that FAF-BSA may act to protect chicken semen and is suitable as a basic component of chicken semen diluent for the method of analyzing rooster semen after freezing.

Demineralized Bone Matrix (DBM) as a Bone Void Filler in Lumbar Interbody Fusion : A Prospective Pilot Study of Simultaneous DBM and Autologous Bone Grafts

  • Kim, Bum-Joon;Kim, Se-Hoon;Lee, Haebin;Lee, Seung-Hwan;Kim, Won-Hyung;Jin, Sung-Won
    • Journal of Korean Neurosurgical Society
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    • v.60 no.2
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    • pp.225-231
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    • 2017
  • Objective : Solid bone fusion is an essential process in spinal stabilization surgery. Recently, as several minimally invasive spinal surgeries have developed, a need of artificial bone substitutes such as demineralized bone matrix (DBM), has arisen. We investigated the in vivo bone growth rate of DBM as a bone void filler compared to a local autologous bone grafts. Methods : From April 2014 to August 2015, 20 patients with a one or two-level spinal stenosis were included. A posterior lumbar interbody fusion using two cages and pedicle screw fixation was performed for every patient, and each cage was packed with autologous local bone and DBM. Clinical outcomes were assessed using the Numeric Rating Scale (NRS) of leg pain and back pain and the Korean Oswestry Disability Index (K-ODI). Clinical outcome parameters and range of motion (ROM) of the operated level were collected preoperatively and at 3 months, 6 months, and 1 year postoperatively. Computed tomography was performed 1 year after fusion surgery and bone growth of the autologous bone grafts and DBM were analyzed by ImageJ software. Results : Eighteen patients completed 1 year of follow-up, including 10 men and 8 women, and the mean age was 56.4 (32-71). The operated level ranged from L3/4 to L5/S1. Eleven patients had single level and 7 patients had two-level repairs. The mean back pain NRS improved from 4.61 to 2.78 (p=0.003) and the leg pain NRS improved from 6.89 to 2.39 (p<0.001). The mean K-ODI score also improved from 27.33 to 13.83 (p<0.001). The ROM decreased below 2.0 degrees at the 3-month assessment, and remained less than 2 degrees through the 1 year postoperative assessment. Every local autologous bone graft and DBM packed cage showed bone bridge formation. On the quantitative analysis of bone growth, the autologous bone grafts showed significantly higher bone growth compared to DBM on both coronal and sagittal images (p<0.001 and p=0.028, respectively). Osteoporotic patients showed less bone growth on sagittal images. Conclusion : Though DBM alone can induce favorable bone bridging in lumbar interbody fusion, it is still inferior to autologous bone grafts. Therefore, DBM is recommended as a bone graft extender rather than bone void filler, particularly in patients with osteoporosis.