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유리화 동결법에 의한 난자의 동결 중 항동해제에 Cytoskeleton Stabilizer, Taxol의 처리 후 배발달율의 개선에 관한 연구

  • 정형민;박성은;임정묵;고정재;정미경;차광렬
    • Proceedings of the KSAR Conference
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    • 2001.03a
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    • pp.74-74
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    • 2001
  • 난자 동결방법의 선별은 보다 효과적인 난자은행의 개발에 필수 불가결한 중요한 요소이다. 이전의 연구에서 마우스의 난자를 ethylene glycol과 electron microscope grid를 이용한 유리화 동결법으로 동결 융해한 결과 기존의 slow freezing 방법에서보다 높은 생존율과 배발달율이 나타남을 관찰하였다. 그러나 동결융해후의 난자는 방추사와 염색체의 이상성이 대조군에 비해 높은 빈도로 나타나 융해후의 배발달율을 감소시키는 것으로 보고되었다. 이에 본 연구에서는 유리화동결법동안 항동해제에 Cytoskeleton system을 안정화시키는 cytoskeleton stabilizer인 taxol을 첨가시킨후 동결시켰을때 생존율과 발달율을 개선시킬 수 있는지 알아보고자 본 실험을 시행하였다. ICR mouse의 성숙란을 채취하여 연구목적에 따라 taxol을 첨가시키지 않은 대조군과 첨가시킨 실험군으로 분류하였다. 동결방법은 난자를 1.5 M ethylene glycol (EG)에 2분 30초간 노출시킨후 5.5 M EG와 1 M sucrose가 첨가된 동결액에 20초간 노출시킨 후 Grid에 난자를 부착시킨후 직접 액체질소에 침지하여 동결하였다. 동결후 난자는 5단계로 융해를 실시한 후 정자와 체외수정을 시킨 후 수정된 난자는 modified P1 배약액에 124 h까지 발달율을 관찰하였고, 배양 후 발달된 배반포는 대조군과 실험군, 각각 4마리의 발정동기화된 recipient에 이식을 시행하였다. 배발달율은 대조군에 비해 실험군에서 4세포기 (48 vs. 84.4%), 8세포기 (34% vs. 70.6%), 상실배 (26% vs. 58.6%) 그리고 배반포 발달율은 (24% vs. 58.6%)로 높게 관찰되었다. 배아이식후 대조군과 실험군에서 각각 2 마리가 임신이되어 정상적인 산자를 분만하였다. 따라서 항동해제에 taxol의 첨가는 동결 융해후의 난자의 배발달율을 증진시킬 수 있었다..8%로 나타나 난할율 및 배반포 발생율에 있어서 융합조건에 따라 큰 차이는 없었으나 1.9㎸/cm, 30$\mu\textrm{s}$ 2회의 조건이 다른 조건들에 비하여 유의적으로 낮았다. 따라서, 체세포와 수핵란 세포질간의 융합율과 배반포 발생에 미치는 영향은 전압보다는 시간에 더 크게 받음을 알 수 있었으며, 이와 같은 결과에서 융합시 시간을 오래 주는 것보다 전압을 높이는 것이 수핵난자의 세포질에 상해를 줄이고 이후 배반포 발생에 유리할 것으로 사료되었다.면에서도 더욱 더 활발할 것으로 기대된다. 배란후 72시간째에 초음파진단기를 이용하여 난소의 난포발달을 조사한 결과 , 대조구와 bFF처리구에 비해 AI처리구에서 발달난포가 유의적으로 많은 것을 확인하였다. 이상과 같은 결과로, Anti-inhibin serum은 한우 자체에서 분비하는 Inhibin을 특이하게 억제하여 Inhibin에 의해 억제되는 FSH분비가 촉진됨으로써 난포발달과 estrogen의 농도가 촉진되는 것으로 사료되어 anti-inhibin serum이 한우의 과배란유기 효과가 있는 것으로 사료된다.정량 분석한 결과이다. 시편의 조성은 33.6 at% U, 66.4 at% O의 결과를 얻었다. 산화물 핵연료의 표면 관찰 및 정량 분석 시험시 시편 표면을 전도성 물질로 증착시키지 않고, Silver Paint 에 시편을 접착하는 방법으로도 만족한 시험 결과를 얻을 수 있었다.째, 회복기 중에 일어나는 입자들의 유입은 자기폭풍의 지속시간을 연장시키는 경향을 보이며 큰 자기폭풍일수록 현저했다. 주상에서 관측된 이러한 특성은 서브스톰 확장기 활동이 자기폭풍의 발달과 밀접한 관계가 있음을 시사한다.se that were all low in two aspects, named "the Nonsignificant group". And the issues were high risk perception in general setting and

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Purification and Properties of Arylsulfatase of Serratia marcescens (Serratia marcens Arylsulfatase의 정제와 성질)

  • Yim, Moo-Hyun
    • Microbiology and Biotechnology Letters
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    • v.5 no.4
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    • pp.177-184
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    • 1977
  • Arylsulfatase catalyzes the release of SO$\sub$4//sup2/- from sulfate esters of simple phenols. Arylsolfatase occurs widely in animal tissues and in microorganisms including soil bacteria. Its widespread distribution suggests that it has a rather fundamental function and environmental meaning. It has been shown previously that arylsulfatase of Klebsiella was purified and characterized. A condition of arylsulfatase synthesis was tested with several strains of Serratia. Serratia marcescens could not utilize some sugars, such as xylose, rhamnose, glucosamine and arabinose hut glucose and mannitol as a sole carbon source. However, arylsulfatase synthesis was repressed by glucose but not by mannitol. The enzyme synthesis was repressed ob inorganic sulfate and methionine, and this repression was relieved by addition of tyramine. Arylsulfatase of S. marcescen was purified by fractionation with ammonium sulfate and followed by chromatographies on DEAE-Cellulose CM-Cellulose, and DEAE-Sephadex A-25. The molecular weight of arylsulfatase was determined to be 46,000 by SDS-Gel electrophoresis and 49,000 by Sephadex G-100 column chromatography. The enzyme showed some different properties with that of K. aerogenes. The activity was maximum at pH 6.8. The Km and Vmax values for p-nitrophenyl sulfate were 2.5${\times}$10$\^$-4/ M and 20 nmoles/min/mg protein, respectively. The enzyme showed high activities toward phenyl sulfate, ο-and p-nitro phenyl sulfates, and p-nitrocatechol sulfate. The inhibition of enzyme was strongly affected by hydroxylamine, inorganic fluoride, sulfide and phosphate, but by inorganic sulfate. Like Klebsiella arylsulfatase, tyramine, octopamine, and dopamine gave signifcant inhibitory effect.

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High Throughput-compatible Screening of Anti-oxidative Substances by Insect Extract Library (약용곤충추출물 라이브러리를 이용한 항산화 활성의 초고속 검색)

  • Park, Ja-Young;Heo, Jin-Chul;An, Sang-Mi;Yun, Eun-Young;Han, Sang-Mi;Hwang, Jae-Sam;Kang, Seok-Woo;Yun, Chi-Young;Lee, Sang-Han
    • Food Science and Preservation
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    • v.12 no.5
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    • pp.482-488
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    • 2005
  • Oxidant stress is well-known for a pivotal parameter related to neuro-inflammatory diseases including Alzheimer's disease, Parkinson's disease, and ALS (Lou Gehrig's disease). In order to effectively screen for anti-inflammatory agents, we first established the infrastructure of high throughput screening for anti-oxidant agents from medicinal insect library extracted with water, methanol, ethanol, and dimethyl sulfoxide. By the screening system, we found that Tenodera angustipennis Saussure, Pyrocoela rupa Olivier and Papilio maackii Mntris had strong anti-oxidant activity. Moreover, Tenodera angustipennis Saussure and Tenodera aridifolia (Stoll) exhibited protection effects of cellular damage by treatment of an oxidant hydrogen peroxide. Together, the results suggest that some selected hits could be a potential agent against neuro-inflammation, although the in vivo studies should be clearly tested.

Inhibition Effect against the Rat Blood Plasma Oxidation of the $Makgeolli$ ($Takju$) Korean Rice Wine (막걸리(탁주)의 쥐 혈장 과산화물 생성 억제효과)

  • Wang, Seung-Jin;Lee, Hyoung-Jae;Cho, Jeong-Yong;Jang, Mi-Young;Park, Keun-Hyung;Moon, Jae-Hak
    • Food Science and Preservation
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    • v.19 no.1
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    • pp.116-122
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    • 2012
  • The antioxidant activities of $makgeolli$ and other alcoholic beverages were compared. Based on the same volume (70 ${\mu}L$ eq.) of the alcoholic beverages, the 2,2'-azinobis (3-ethylbenzothiazoline-6-sulfonic acid ammonium salt) (ABTS+) radical-scavenging activities were as follows: whisky > $makgeolli$ crude extract (MCE) > rice wine (RW) > clarified $makgeolli$ (CM) > soju. Based on the same alcohol concentration (6%) of the alcoholic beverages, however, $makgeolli$ showed the highest activity. In addition, based on the same volume (70 ${\mu}L$ eq.), the inhibition effects against the formation of cholesteryl ester hydroperoxide (CE-OOH) were as follows: soju > whisky > RW > MCE > CM. Based on the same alcohol concentration (6%), however, the inhibition effects against the formation of CE-OOH were as follows: RW > MCE > soju > whisky > CM. Therefore, it was suggested that $makgeolli$ may contain radical-scavenging- and metal-ion-chelate-type antioxidants and may increase the antioxidant activity in the blood.

Variation of Bolting at Cultivation of Different Regions and Molecular Characterization of FLC homologs in Angelica gigas Nakai (재배 지대에 따른 참당귀의 추대 변이와 FLC 유전자 특성)

  • Kim, Young-Guk;Yeo, Jun-Hwan;An, Tae-Jin;Han, Sin-Hee;Ahn, Young-Sup;Park, Chung-Beom;Jang, Yun-Hee;Kim, Jeong-Kook
    • Korean Journal of Medicinal Crop Science
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    • v.20 no.5
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    • pp.359-364
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    • 2012
  • This study were carried out to find bolting response of cultivation in different regions and to isolate FLC (FLOWERING LOCUS C) homologs in Angelica gigas Nakai. The mean temperature of different regions, ordering in altitude, were as follows: 100 m > 350 m > 530 m > 700 m. The largest amount of rainfall was occurred in the region of 350 m while the longest time of sunshine was occurred in the region of 100 m. The content of soil chemical properties in regions showed pH 6.2 ~ 7.4, T-N 0.17 ~ 26, organic mater $1{\sim}32gkg^{-1}$, $P_2O_5$ ${151{\sim}664_{mgkg}}^{-1}$, exchangeable potassium and calcium and magnesium were 0.78 ~ 1.15, 3.9 ~ 10.0, ${0.7{\sim}3.2_{cmol}}^{+kg-1}$. L5 line of A. gigas was occurred in bolting at all regions, but the bolting ratio was 60.0% in 700 m region with non-mulching treatment. Manchu of A. gigas was not occurred in bolting at all regions. The accumulation bolting ratio of L5 line by non-mulching was higher than that of mulching as 90.4% and 72.8% in 100 m region. The MADS-box transcription factor FLC is one of the well-known examples as a strong floral repressor. We decided to isolate FLC homologs from A. gigas as a starting point of flowering mechanism research of this plant. We have isolated two RT-PCR products which showed very high amino acid sequence homology to Arabidopsis FLC.

Geophysical and Geochemical Studies for the Saline Water Intrusion under the Paddy Field in Kyoung-gi area, Korea (경기 지역 농경지 하부로의 해수 침투에 관한 지구물리 및 지구화학적 연구)

  • Lee Sang-Ho;Kim Kyoung-Woong;Lee Sang-Kyu
    • Geophysics and Geophysical Exploration
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    • v.2 no.2
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    • pp.96-103
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    • 1999
  • Most of saline water inousions have been diagnosed by geophysical or geochemical approach independently. The objective of this study is to provide the effective method to detect the saline water intrusion on the ground water in the vicinity of seashore using these two methods. Schulumberger sounding, frequency domain electromagnetic sounding and geochemical analysis of ground water were carried out to explore saline water intrusion. Schulumberger sounding was implemented in dry surface condition before irrigation water was introduced into the field, while electromagnetic sounding was carried out in wet ground condition after the irrigation. The purpose of duplicated measurements on the equivalent spot at different times was to investigate the variation of anomaly zone depending on the amount of ground water. It was possible to discriminate the anomalous zone due to high water saturation from the low electric resistivity zone by high salt concentrations through this way. For the verification of the geophysical result, the ground water samples in the study area were collected and analysed at the 23 points near the measuring spots. The groundwater at the spot nearest to the sea water intrusion identified by geophysical method indicates higher salinity than the standard limit concentration for agricultural irrigation water (250 mg/1). Isotope analysis of $D({^2}H)$ vs. is ${^18}O$ and PCA analysis were used to discriminate the anthropogenic pollution from those of high salinity from sea water intrusion.

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Antifungal Activity of Bacillus sp. BCNU 2003 against the Human Pathogenic Fungi (인체 병원성 진균에 대한 Bacillus sp. BCNU 2003의 항진균 효과)

  • Choi, Hye-Jung;Yang, Uk-Hee;Kim, Ya-Ell;Choi, Yeon-Hee;Ahn, Cheol-Soo;Jeong, Young-Kee;Kim, Dong-Wan;Joo, Woo-Hong
    • Journal of Life Science
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    • v.20 no.2
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    • pp.269-274
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    • 2010
  • An antifungal antibiotic-producing strain, BCNU 2003, was isolated from forest soil in Korea. The morphological and physiological characters, and 16S rRNA sequences analysis of strain BCNU 2003 identified this strain as Bacillus genus. The Bacillus sp. BCNU 2003 showed strong antifungal activities against Aspergillus niger, Trichophyton mentagrophytes and Trichophyton rubrum with inhibition ranging from 62.05 to 63.49% by using dual culture technique. Bacillus sp. BCNU 2003 produced a maximum level of antifungal substances under aerobic incubation at 28oC and pH 6.5-7.2 for 6 days in LB broth. Ethyl acetate extract of the cultured broth showed strong antifungal activity and a broad antifungal spectrum against various pathogenic fungi. The minimum inhibitory concentration (MIC) values for its active extracts ranged between 0.0625 mg/ml and 1 mg/ml. In addition, Bacillus sp. BCNU 2003 was determined to have the ability to produce enzymes such as amylase, protease, gelatinase and catalase.

PEP-1-GSTpi protein enhanced hippocampal neuronal cell survival after oxidative damage

  • Sohn, Eun Jeong;Shin, Min Jea;Kim, Dae Won;Son, Ora;Jo, Hyo Sang;Cho, Su Bin;Park, Jung Hwan;Lee, Chi Hern;Yeo, Eun Ji;Choi, Yeon Joo;Yu, Yeon Hee;Kim, Duk-Soo;Cho, Sung-Woo;Kwon, Oh Shin;Cho, Yong-Jun;Park, Jinseu;Eum, Won Sik;Choi, Soo Young
    • BMB Reports
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    • v.49 no.7
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    • pp.382-387
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    • 2016
  • Reactive oxygen species generated under oxidative stress are involved in neuronal diseases, including ischemia. Glutathione S-transferase pi (GSTpi) is a member of the GST family and is known to play important roles in cell survival. We investigated the effect of GSTpi against oxidative stress-induced hippocampal HT-22 cell death, and its effects in an animal model of ischemic injury, using a cell-permeable PEP-1-GSTpi protein. PEP-1-GSTpi was transduced into HT-22 cells and significantly protected against H2O2-treated cell death by reducing the intracellular toxicity and regulating the signal pathways, including MAPK, Akt, Bax, and Bcl-2. PEP-1-GSTpi transduced into the hippocampus in animal brains, and markedly protected against neuronal cell death in an ischemic injury animal model. These results indicate that PEP-1-GSTpi acts as a regulator or an antioxidant to protect against oxidative stress-induced cell death. Our study suggests that PEP-1-GSTpi may have potential as a therapeutic agent for the treatment of ischemia and a variety of oxidative stress-related neuronal diseases.

Epidemiological change on Paragonimiasis 2nd intermediate host in Kang Hwa Gun (강화군(江華郡)의 폐흡충중간숙주(肺吸虫中間宿主)에 관(関)한 역학적(疫學的) 추이(推移))

  • Lee, J.S.;Kim, J.R.;Kim, S.J.;Rim, H.J.;Song, O.D.;Kim, M.S.
    • Journal of agricultural medicine and community health
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    • v.4 no.1
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    • pp.71-80
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    • 1979
  • Paragonimiasis is a common trematodiasis of medical importance and one of public health problems in many areas of sob thern part of Korea. During the last sixty years, several on the prevalence among the Korean population have been carried out by various investigators, although the general distribution and prevalences of paragonimiasis are now yet clearly determind. This study was undertaken to evaluate the status of human paragonimiasis in Kang Hwa Gun, Kyung gi Province, Korea by the use of intradermal screening test whih Veronal buffered saline antigen. A total number of 208 residents (male 119, female 89) of all age group in Kilgeong-Ri, Yangdo Myun, Kang Hwa Gun were examined by the intradermal test with paragonimus antigen. The infestation rate of crayfish with Paragonimus metacerearia was also examine in Konneung and Jeoungha village, Kilgeong-Ri from March 1973 to October 1978 by the interval of two to five years. 1. In the resnlt of 208 residents, Kilgeong-Ri, the positive rate showed in 29.3% in as a whole(36.1% in male, 20.2% in female). The highest positive rate showed in 10 to 19 age group(41.5%). 2. Among the residents in Kilgeong-Ri, Paragonimus eggs were detected in 11 residents by the sputum and the stool examinations without the intradermal test result. 3. The infestation rate of crayfish with Paragonimus inetacercaria was 90.6%(116 out of 128 crayfishes) in Konneung village, and 61.5%(24 out of 39) crayfishes) in Jeongha village, Kilgeng-Ri. 4. The fluctation of the infestation tate and density of crayfish with Paragonimus metacercaria was decreased by the interval of two to five years in Konneung and Jeongha village in Kilgeong-Ri.

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Simultaneous Determination of Chromium (III) and Chromium(VI) by High Performance Liquid Chromatography(HPLC) (고성능 액체크로마토그래피(HPLC)를 이용한 3가, 6가 크롬의 동시정량에 관한 연구)

  • Roh, Jae Hoon;Kim, Chi Nyon;Kim, Choon Sung;Kim, Kyoo Sang
    • Journal of Korean Society of Occupational and Environmental Hygiene
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    • v.4 no.2
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    • pp.189-197
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    • 1994
  • Analytic methods for Cr(VI) level in industrial hygienic field were suggested by the National Institute for Occupational Safety and Health(NIOSH method 7600, 7604). There were growing needs for measurement of Cr(III) and Cr(VI) levels simultaneously. Two analytical methods were suggested to determine Cr(III) and Cr(VI) levels simultaneously. The one is method by using reversed phase high peformance liquid chromatography(HPLC) and the other is by using ion exchange HPLC. The purpose of this work was to evaluate the usefulness of these two analytic methods. For the difference of ionic charges of Cr(III)-ethylendiamine tetraacetic acid(EDTA) chelate and $CrO_4{^-2}$, we could detect them simultaneously by ion exchange HPLC. Also, we attempted to determine the levels of Cr(III) and Cr(VI) chelated with sodium diethyldithiocarbamate(NaDDTC) by using reversed phase HPLC. The confirmation of Cr(III) and Cr(VI) were checked by fraction collector and nameless atomic absorption spectrometer. The optimal conditions for the formation of Cr(III)-EDTA chelate were two hours incubation period with pH 5. Cr(III)-EDTA and Cr(VI) in EDTA solution were successfully separated by anion exchange column using $Na_2CO_3/NaOH$ mixture as mobile phase. Peaks of Cr(III)-EDTA and Cr(VI) in EDTA were identified at 5 minutes and 7 minutes of retention time respectively by the ion exchange HPLC. The formation of Cr(III)-NaDDTC and Cr(VI)-NaDDTC chelates were twelve hours incubation period. Cr(III)-NaDDTC and Cr(VI)-NaDDTC chelates were separated by reversed phase column using methanol and water mixture as mobile phase. Peaks of Cr(VI)NaDDTC and Cr(III)-NaDDTC chelates were identified at 13 minutes and 26 minutes of retention time respectively by the reversed phase HPLC. Due to reduction of Cr(VI) to Cr(III), it seems to be not suitable for simultaneous determination of Cr(III)-NaDDTC and Cr(VI)-NaDDTC chelates by reversed phase HPLS. Simultaneos determination of Cr(III) and Cr(VI) by ion exchange HPLC was more accurate and simple method.

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