• 한국환경과학회지
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• 제19권10호
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• pp.1307-1314
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• 2010

This study was conducted to isolate and characterize a novel feather-degrading bacterium producing keratinase activity. A strain K9 was isolated from soil at poultry farm and identified as Xanthomonas sp. K9 by phenotypic characters and 16S rRNA gene analysis. The cultural conditions for the keratinase production were 0.3% fructose, 0.1% gelatin, 0.04% $K_2HPO_4$, 0.06% $KH_2PO_4$, 0.05% NaCl and 0.01% $FeSO_4$ with an initial pH 8.0 at $30^{\circ}C$ and 200 rpm. In an optimized medium containing 0.1% chicken feather, production yield of keratinase was approximately 8-fold higher than the yield in basal medium. The strain K9 effectively degraded chicken feather meal (67%) and duck feather (54%), whereas human nail and human hair showed relatively low degradation rates (13-22%). Total free amino acid concentration in the cell-free supernatant was about 25.799 mg/l. Feather hydrolysate produced by the strain K9 stimulated growth of red pepper, indicating Xanthomonas sp. K9 could be not only used to increase the nutritional value of chicken feather but also a potential candidate for the development of natural fertilizer applicable to crop plant soil.

• Bulletin of the Korean Chemical Society
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• 제10권5호
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• pp.472-474
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• 1989

• 식물병연구
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• 제11권2호
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• pp.158-161
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• 2005

결구상추 밑둥썩음병원균인 Rhizoctonia. solani에 대한 길항세균으로 보고된 Stenotrophomonas maltophilia BW-l3 균주의 배양적 특성과 항균물질의 생산능을 조사하고 대량배지를 선발하였다. BW-13 균주의 초기 pH와 배양적온을 조사한 결과, pH는 $6.0\~8.0$에서 , 온도는 $35^{\circ}C$에서 세균의 생육이 가장 높았다. 대량배양을 위한 효율적인 탄소원을 선발하기 위하여 glucose, sucrose, lactose, maltose, manose 등의 저분자 탄소원과 개량제, 미강, 옥수수전분, 고구마전분 등의 고분자 탄소원을 사용하여 BW-13균주의 생장과 항균활성에 미치는 영향을 조사하였다. 그 결과, 기초배지에 $3\%$개량제를 첨가했을 때 BW-13 균주의 세균 증식과 항균활성에 가장 효과적이었다. 따라서 기초배지($1.25\%\;K_{2}HPO_4,\;0.38\%\;KH_{2}PO_4,\;0.01\%\;MgSO_4{\cdot}7H_{2}O,\;0.5\%\;Yeast extract$)에 개량제 $3\%$ 첨가한 배지(dough conditioner media)를 BW-13 균주의 대량배양용 배지로 선발하였다.

• 한국구조물진단유지관리공학회 논문집
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• 제20권5호
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• pp.93-101
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• 2016

본 연구에서는 비국지 탄성이론과 미분변환법을 이용하여 탄성매질속 다중 크랙을 가진 비균질 나노빔의 지배방정식을 유도하고, 유도된 지배방정식과 경계조건에 미분변환법을 적용하여 나노빔의 축방향의 진동해석을 하며, 나노빔의 첫단과 끝단의 경계조건이 각각 고정단(Clamped end)과 자유단(Free end)의 경우에 대하여 수치해석을 수행하였다. 수치해석 결과를 기존 연구결과와 비교하여 타당성을 입증한 후, 비국지 작은 척도효과 (Nonlocal small scale effect), 탄성매질의 강성, 크랙의 위치, 크랙의 강성 그리고 비국지 탄성이론의 나노빔에 대한 진동해석 결과를 고찰하였다.

• Clinical and Experimental Reproductive Medicine
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• 제30권4호
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• pp.269-280
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• 2003

Objective: We reported the overcoming effect of $Ni^{2+}$ on the in vitro 2-cell block of mouse embryos. In this study, we aim to investigate whether $Ni^{2+}$ should induce intracellular $Ca^{2+}$ transient in the mouse embryos. Materials and Methods: Embryos were collected at post hCG 32hr from the oviduct of the ICR mouse and cultured in M2 medium omitted phenol red. Intracellular $Ca^{2+}$ was checked by using a confocal laser scanning microscope and fluo-3AM by using various intracellular $Ca^{2+}$ antagonists. Results: In 1mM $Ni^{2+}$ treated medium which contained $Ca^{2+}$(1.71mM), 75.7% of the embryos showed $[Ca^{2+}]i$ transient about 200 sec later. In the $Ca^{2+}$-free medium, 69.8% of the embryos showed $[Ca^{2+}]i$ transient. In U73122, phospholipaseC(PLC) inhibitor (5uM, 10min) pretreated group, 33.3% of the embryos showed $[Ca^{2+}]i$ transient. Heparine, inositol 1, 4, 5-triphosphate receptor(IP3R) antagonist preinjected embryos showed no response with 1mM $Ni^{2+}$. In danthrolene treatment, ryanodine receptor(RyR)-antagonist, 43% embryos showed $[Ca^{2+}]i$ transient but they showed delayed response about 340sec in the presence of $Ca^{2+}$. Conclusions: Summing up the above results, $Ni^{2+}$ seems to induce $Ca^{2+}$-release from the $Ca^{2+}$-store even in the $Ca^{2+}$-free medium. IP3 receptors of the mouse 2-cell embryos might have an essential role for the intracellular $Ca^{2+}$ increase by $Ni^{2+}$.

• Journal of Animal Science and Technology
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• 제57권8호
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• pp.27.1-27.6
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• 2015

This study was designed to investigate the effects of ${\alpha}$-tocopherol and granulosa cells monolayer on nuclear maturation and cleavage rates of ovine cumulus-oocyte complexes (COCs). The COCs (n = 2814) were matured in maturation medium supplemented with various concentration of ${\alpha}$-tocopherol (0, 5, 10, $15{\mu}g/ml$), oocytes were incubated at $39^{\circ}C$ with 5 % $CO_2$ for 24 h in three culture systems: (a) maturation medium (MM; n = 884), (b) co-cultured with granulosa cells (CG; n = 982) and (c) co-cultured with granulosa cells and cells were further cultured in MM for 12 h (CG + 12hMM; n = 948). Our results showed that ${\alpha}$-tocopherol had no effect on GVBD and MII as compared to control group, but when ${\alpha}$-tocopherol added to maturation medium the rate of cleavage decreased. This indicates interaction of above mentioned factors in any of the treatments showed no significant differences on the rate of maturation and cleavage stages (MII, GVBD and cleavage) (p > 0.05). The oocytes co-cultured with granulosa cells for 24 h had beneficial effects on cleavage rate. The maximum MII and cleavage rates were achieved when oocytes had extra 12 h culture in the maturation medium without granulosa cells. Results also showed our modified co-culture system (CG + 12hMM), improved rates of MII and the cleavage in comparison with other studied maturation systems.

• Journal of Microbiology and Biotechnology
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• 제1권2호
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• pp.142-149
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• 1991

Gardenia callus was induced in MS medium containing $10{\;}{\mu}M$ of 2,4 diphenoxy acetic acid (2,4-D), $1{\;}{\mu}M$ kinetin, and 3% sucrose in the dark. $B_5$ medium was identified to be the most adequate medium for cell growth. Indole-3-acetic acid (IAA) was better growth regulator than 2,4-D not only for cell growth but slso for carotenoid production. Ligt also played a critical role on synthesis of carotenoid. Gardenia cells grown in $B_5$ medium could utilize a polysaccharide, soluble starch, as a carbon source. The cell growth was stimulated in $B_5$ medium fortified with 0.2% yeast extract. The optimum pH for cell growth was 5.7. High density cultures can be maintained by increasing inoculum size and medium concentration accordingly. Specific growth rate and mass doubling time were 0.095 $day^{-1}$ and 7.3 days, respectively. The cell immobilized in alginate tends to formulate more enlarged vacuoles containing yellow pigment compared with those of suspended cell. Carotenoid content of immobilized cell was about $264.4{\;}{\mu}g/g$ fresh weight (F.W.) corresponding twice of the content of suspended cell ($112.08{\;}{\mu}g/g$ F.W.). The color of gardenia cell was shifted from yellow to red when carbohydrase-secreting fungus, Trichoderma reesei, was co-cultivated with gardenia cells.

• Journal of Microbiology and Biotechnology
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• 제1권2호
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• pp.116-120
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• 1991

The transformant Bacillus subtilis ED213 carrying the pSO100 which cloned the cdd gene encoding cytidine deaminase (cytidine /2'-deoxycytidine aminohydrolase, EC 3.5.4.5, CDase) originated from wild type B. subtilis was cultivated in Spizizen minimal medium (SMM). To overcome poor expression of the cdd gene in SMM medium, the medium compositions and growth conditions were optimized. The optimized medium compositions and growth conditions were cytidine concentration of 80 mg/l, glycerol of 25 g/l, and $(NH_4)_2SO_4$ of 10 g/l, along with $37^{\circ}C$ and pH 7.0. The intracellular CDase production was increased 3 times from 1,000 unit/ml to 3,200 unit/ml, and extracellular CDase also increased from nearly undetectable amounts to 1,500 unit/ml. The cytidine concentration was signified as the most critical compositional factor for overproduction of CDase by increasing the cell density mainly in culture broth. The plasmids were more stable in cells that were grown in original SMM medium with stability of 90% compared to those grown in optimized SMM medium with stability of 80% after 48 hours cultivation. The most active amplification of plasmid was occurred in the logarithmic phase, which showed a value around four times higher than the initial copy number. In the exponential phase, the CDase production was closely related to the plasmid copy number along with the cell density. However, it was not accorded with cell density at the stationary phase.

• Clinical and Experimental Reproductive Medicine
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• 제27권3호
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• pp.275-282
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• 2000

Objective: This study was to determine the effect of different concentration of calcium III medium on the preimplantational development of zygotes and early 2 cell embryos. Methods: Female mice of ICR strain ($5{\sim}8$ weeks old) were superovulated and mated with fertile males. Zygotes or early 2-cell embryos were collected by flushing the oviducts $31{\sim}32$ hours after hCG injection. The embryos were cultured in various concentrations of $Ca^{2+}$ in medium or with EDTA, EGTA and $Ni^{2+}$. Result and Conclusion: Treatment of high concentraion of $Ca^{2+}$ (3.42 mM $(2X){\sim}17.l$ mM (10X)) in medium didn't develop well compared to the control. Low concentrations of $Ca^{2+}$ (0.214mM $(1/8X){\sim}0.855$ mM (1/2X) were deterimental to development beyond 2-cell stage. EDTA, $Ca^{2+}$ chelating agent was treated with ranged concentrations of EDTA (0.014 $mM{\sim}0.107$ mM) to medium contaning 1.71 mM $Ca^{2+}$ showed beneficial effect to development to blastocyst compared to the control. EGTA, extracellular $Ca^{2+}$ chelator, was treated with ranged concentrations of EGTA ($0.014{\sim}0.107$ mM) to the medium contaning 1.71 mM $Ca^{2+}$. There is no significant difference with the control. $Ni^{2+}$ (50 ${\mu}M$), T-type $Ca^{2+}$-channel blocker was treated to medium contaning low concentration of $Ca^{2+}$. It overcame 2-cell block significantly. Rate of degenerated embryos decreased and developmental rate to morula and blastocyst increased more than low $Ca^{2+}$ concentration alone. Further studies are needed for the overcoming effect of 2-cell block by $Ni^{2+}$.

• 생태와환경
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• 제45권3호
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• pp.271-277
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• 2012

본 연구에서는 D. magna의 배양배지로서의 국내 자연수의 적절성을 평가하기 위하여, Elendt M4 배지에서 생산된 태어난 지 24시간 미만 된 어린 D. magna를 Elendt M4 배지, 탈염소 수도수 및 먹는 샘물에 21일 동안 노출시켜 생존율 및 번식능력을 평가하였다. 대조배지인 Elendt M4 배지와 먹는 샘물에서 배양한 D. magna는 어미의 생존율, 첫 배를 생산하는 시기, 생존한 어미 당 생산된 총 어린 물벼룩 평균수, 생존한 어미 당 생산된 죽은 어린 물벼룩 평균수는 2회의 번식시험 모두에서 Jonczyk과 Gilron (2005) 및 OECD No. 211, Daphnia magna Reproduction Test 지침서(OECD, 2008)의 기준을 벗어나지 않았다. 그러나 탈염소 수도수에서 배양을 한 경우에는 2번의 번식시험 모두 어미의 사망률이 20% 이상으로, 배양 13일, 15일, 18일에 사망된 개체가 관찰되었다. D. magna는 경도가 80 mg $CaCO_3\;L^{-1}$ 이상인 물에서 사용을 추천하고 있으나, 본 연구에서 사용된 탈염소 수도수의 경도는 50~53 mg $CaCO_3\;L^{-1}$ 이었다. 탈염소 수도수에서 나타난 지연된 사망률은 배양배지의 급격한 경도 차이에 의한 영향으로 판단된다. 그러므로 국내의 자연수(지하수, 표면수, 탈염소 수도수 등)를 사용하여 D. magna를 배양할 경우, 배양배지의 경도를 100 mg $CaCO_3\;L^{-1}$ 이상 강화시켜 사용하는 것이 필요하다. 그리고 궁극적으로는 국내에 서식하는 토착 물벼룩류를 대상으로 국내 수 환경에 적합한 시험생물을 개발하는 국가적인 연구가 필요하다고 사료된다.