• 제목/요약/키워드: Artificial incubation

검색결과 142건 처리시간 0.026초

고분자필름과 금속막 의료소재에 대한 생체적합성 및 독성 평가를 위한 새로운 세포배양시스템의 개발 및 적용 (Development and Application of a Novel Mammalian Cell Culture System for the Biocompatibility and Toxicity of Polymer Films and Metal Plate Biomaterials)

  • 곽문화;윤우빈;김지은;성지은;이현아;서은지;남국일;정영진;황대연
    • 생명과학회지
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    • 제26권6호
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    • pp.633-639
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    • 2016
  • 고분자(polymer), 금속(metal), 세라믹(ceramic), 합성물(composite) 등과 같은 바이오소소재(Biomaterials)는 그들의 물리화학적 성질 때문에 의료용섬유(medical fibers), 인공혈관(artificial blood vessels), 인공관절(artificial joints), 임플란트(implants), 연조직(soft tissue), 인공성형물(plastic surgery materials) 등 의료용으로 많이 사용되며, 개발연구도 활발히 진행되고 있다. 그러나, 필름(film)이나 판(plate)형태의 바이오소재에 대한 생체적합성(biocompatibility)이나 독성(toxicity)을 포유동물세포를 이용하여 평가하는 것은 적절한 평가용 장치가 없기 때문에 매우 어려운 상황이다. 따라서 이러한 문제를 해결하기 위해서, 본 연구에서는 고분자필름이나 금속판에 유용하게 적용할 수 있는 실리콘링, 상판(top panel), 하판(bottom panel)으로 구성된 새로운 포유동물배양시스템을 개발하고, 이를 실제 적용하고자 하였다. 개발된 시스템은 평가하고자 하는 시료를 상판과 하판사이에 조립하는 샌드위치시스템을 기반으로 한다. 세포배양장치의 조립 후, SK-MEL-2세포를 3가지 시료; Styela Clava Tunic (SCT)- PF, NaHCO3-added SCT (SCTN)-PF, magnesium MP (MMP)에 적용하고 37℃ 이산화탄소 배양기에서 24시간과 48시간 동안 배양하였다. MTT분석결과에서, 세포생존율(cell viability)은 24시간과 48시간 동안 SCT-PF배양그룹에서 정상적으로 유지되었지만 48시간 동안 SCTN-PF배양그룹에서는 급격하게 감소되었다. 더불어, MMP배양그룹에서 세포생존율은 24시간과 48시간 배양 후에 대조군과 유사하게 유지되었다. 이러한 결과는 본 연구에서 새롭게 개발된 샌드위치형태의 포유동물세포배양장치는 고분자필름이나 금속판형태의 바이오소재에 대한 독성이나 생체적합성을 평가하기 위한 우수한 잠재력을 보유하고 있음을 제시하고 있다.

Effects of Addition Level and Chemical Type of Propionate Precursors in Dicarboxylic Acid Pathway on Fermentation Characteristics and Methane Production by Rumen Microbes In vitro

  • Li, X.Z.;Yan, C.G.;Choi, S.H.;Long, R.J.;Jin, G.L.;Song, Man K.
    • Asian-Australasian Journal of Animal Sciences
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    • 제22권1호
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    • pp.82-89
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    • 2009
  • Two in vitro experiments were conducted to examine the effects of propionate precursors in the dicarboxylic acid pathway on ruminal fermentatation characteristics, $CH_4$ production and degradation of feed by rumen microbes. Fumarate or malate as sodium salts (Exp. 1) or acid type (Exp. 2) were added to the culture solution (150 ml, 50% strained rumen fluid and 50% artificial saliva) to achieve final concentrations of 0, 8, 16 and 24 mM, and incubated anaerobically for 0, 1, 3, 6, 9 and 12 h at $39^{\circ}C$. For both experiments, two grams of feed consisting of 70% concentrate and 30% ground alfalfa (DM basis) were prepared in a nylon bag, and were placed in a bottle containing the culture solution. Addition of fumarate or malate in both sodium salt and acid form increased (p<0.0001) pH of culture solution at 3, 6, 9 and 12 h incubations. The pH (p<0.0001) and total volatile fatty acids (VFA, p<0.05) were enhanced by these precursors as sodium salt at 3, 6 and 9 h incubations, and pH (p<0.001) and total VFA (p<0.01) from fumarate or malate in acid form were enhanced at a late stage of fermentation (9 h and 12 h) as the addition level increased. pH was higher (p<0.001) for fumarate than for malate as sodium salt at 3 h and 6 h incubations. Propionate ($C_3$) proportion was increased (p<0.0001) but those of $C_2$ (p<0.05) and $C_4$ (p<0.01 - p<0.001) were reduced by the addition of sodium salt precursors from 3 h to 12 incubation times while both precursors in acid form enhanced (p<0.011 - p<0.0001) proportion of $C_3$ from 6h but reduced (p<0.018 - p<0.0005) $C_4$ proportion at incubation times of 1, 3, 9 and 12 h. Proportion of $C_3$ was increased (p<0.05 - p<0.0001) at all incubation times by both precursors as sodium salt while that of $C_3$ was increased (p<0.001) from 6h but $C_4$ proportion was decreased by both precursors in acid form as the addition level increased. Proportion of $C_3$ was higher (p<0.01 - p<0.001) for fumarate than malate as sodium salt from 6 h incubation but was higher for malate than fumarate in acid form at 9 h (p<0.05) and 12 h (p<0.01) incubation times. Increased levels (16 and 24 mM) of fumarate or malate as sodium salt (p<0.017) and both precursors in acid form (p<0.028) increased the total gas production, but no differences were found between precursors in both chemical types. Propionate precursors in both chemical types clearly reduced (p<0.0001 - p<0.0002) $CH_4$ production, and the reduction (p<0.001 - p<0.0001) was dose dependent as the addition level of precursors increased. The $CH_4$ generated was smaller (p<0.01 - p<0.0001) for fumarate than for malate in both chemical types. Addition of fumarate or malate as sodium type reduced (p<0.004) dry matter degradation while both precursors in both chemical types slightly increased neutral detergent fiber degradability of feed in the nylon bag.

Synthesis and In-vitro Evaluation of N4-Amino Acid Derivatives of Cytarabine for Improving the Oral Delivery of Cytarabine

  • Jin, Ming-Ji;Hong, Joon-Hee;Han, Hyo-Kyung
    • Journal of Pharmaceutical Investigation
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    • 제38권4호
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    • pp.255-259
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    • 2008
  • The present study aimed to investigate the in-vitro characteristics of N4-amino acid derivatives of cytarabine for the oral delivery of cytarabine. After the synthesis of L-Ile-cytarabine, L-Leu-cytarabine and L-Arg-cytarabine, the gastrointestinal stability of each prodrug was examined using artificial gastric juice and intestinal fluids. The cellular uptake characteristics of prodrugs were also examined in Caco-2 cells. While L-Ile-cytarabine and L-Leu-cytarabine appeared to be stable in all the tested biological media during 4-hr incubation, L-Arg-cytarabine was rapidly disappeared within 5 min. Accordingly, the cellular uptake of L-Ile-cytarabine and L-Leu-cytarabine was significantly higher than that of its parent drug, cytarabine in Caco-2 cells but the cellular uptake of L-Arg-cytarabine was similar to that from its parent drug. The cellular uptake of L-Ile-cytarabine and L-Leu-cytarabine appeared to be saturable as drug concentration increased from 0.4 to 4 mM. Collectively, L-Ile-cytarabine and L-Leu-cytarabine could be promising candidates to improve the oral absorption of cytarabine via a saturable transport pathway.

장수상황버섯(Phellinus baumii)의 병재배 특성 (Cultural Characteristics of Phellinus baumii Grown in Bottle)

  • 류영현;조우식;이진만;김종국
    • 한국균학회지
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    • 제32권2호
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    • pp.101-104
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    • 2004
  • 장수상황버섯(Phellinus baumii)은 현재 원목재배가 주종을 이루는 있는데 자실체 생육기간이 길고 원목수급문제가 발생할 가능성이 있고 톱밥의 경우 목재부산물로 다량 발생하고 있어 이를 활용한 톱밥병재배을 실시하였다. 장수상황버섯(Phellinus baumii)의 톱밥병재배를 실시한 결과 참나무톱밥 + 미강(4 : 1)배지가 배양속도와 균사밀도는 가장 높았지만 자실체를 형성하지 못하였고 반면에 참나무톱밥 100% 배지는 균사밀도와 배양속도는 낮았지만 병배지에서 자실체를 형성할 수 있었고 850 ml병에 비해서 1,100 ml 병 처리가 수량성과 생체효율이 약간 높은 결과가 나타났다.

Phelligridin D from Phellinus baumii Reduces Boar Sperm Viability

  • Yi, Young-Joo;Lee, In-Kyoung;Seo, Dong-Won;Yun, Bong-Sik;Lee, Sang-Myeong
    • 한국균학회지
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    • 제44권2호
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    • pp.122-125
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    • 2016
  • Phelligridin D (Phe D) is a compound isolated from Phellinus baumii, which is known for various biological activities. In this study, the authors examined the effect of Phe D on boar spermatozoa for its potential application in assisted reproductive technology for mammals. Sperm motility and deubiquitinylating activity significantly decreased when boar spermatozoa were incubated with Phe D (>$0.5{\mu}M$). The fluorescence intensities of dead sperm, and reactive oxygen species production increased after sperm incubation in the presence of Phe D. Although Phe D is associated with antioxidant and free radical scavenging activity, sperm viability deteriorated after its addition. This could lead to fertilization failure, including that following artificial insemination or in vitro fertilization. Phe D might have other biological functions in spermatozoa, and therefore requires additional studies in the future.

월년잠종의 인공부화에 관한 연구 (Studies on Artificial Hatching of Hibernating Eggs, Bombyx mori L.)

  • 김윤식;손해용
    • 한국잠사곤충학회지
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    • 제18권1호
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    • pp.7-10
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    • 1976
  • 월년종의 냉장처리일수 및 인공부화에 따른 부화상태를 알고자 춘기채종 잠 103$\times$잠 104를 공시하여 1월하순에서 2월 중순에 걸친 시험에서 1월 하순경에 잠난은 휴면타파가 이루어 졌으나 5$^{\circ}C$ 냉장기간이 길어짐에 따라 휴면이 완전 타파되어 최청이 가능하다고 생각한다. 인공난화처리는 영기 비중 1.075 보다는 1.10이 변화에 좋은 영향을 미쳤으며 침지시간은 2분보다는 3분이 효과적이었다. 저온처리와 인공부화처리간에는 저온처리가 짧은 경우는 인공난화처리로서 부화에 효과적이나 반대로 긴 경우는 인공부화 양리량이 강할 수록 난 영산농도와 침지시간이 강할 수록 효과적이었다.

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Role of Non-Thermal DBD Plasma on Cell Migration and Cell Proliferation in Wound Healing

  • Ali, Anser;Lee, Seung Hyun;Kim, Yong Hee;Uhm, Han Sup;Choi, Eun Ha;Park, Bong Joo
    • 한국진공학회:학술대회논문집
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    • 한국진공학회 2013년도 제44회 동계 정기학술대회 초록집
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    • pp.526-526
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    • 2013
  • Plasma technology isbeing developed for a range of medical applications including wound healing. However, the effect of plasma on many cells and tissues is unclear. Cell migration and cell proliferation are very important biological processes which are affected by plasma exposure and might be a potential target for plasma therapy during wound healing treatment. In this study, we confirmed the plasma exposure time and incubation time after plasma treatment in skin fibroblast (L-929 cells) to evaluate the optimal conditions forplasma exposure to the cell in-vitro. In addition, we used a scratch method to generate artificial wound for evaluating the cell migration by plasma treatment. Where, the cells were treated with plasma and migration rate was observed by live-cell imaging device. To find the cell proliferation, cell viability assay was executed. The results of this study indicate the increased cell proliferation and migration on mild plasma treatment. The mechanisms for cell migration and cell proliferation after plasma treatment for future studies will be discussed.

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Hexavalent Chromium Reduction by Bacteria from Tannery Effluent

  • Batool, Rida;Yrjala, Kim;Hasnain, Shahida
    • Journal of Microbiology and Biotechnology
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    • 제22권4호
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    • pp.547-554
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    • 2012
  • Chromium is generated from several industrial processes. It occurs in different oxidation states, but Cr(III) and Cr(VI) are the most common ones. Cr(VI) is a toxic, soluble environmental contaminant. Some bacteria are able to reduce hexavalent chromium to the insoluble and less toxic Cr(III), and thus chromate bioremediation is of considerable interest. An indigenous chromium-reducing bacterial strain, Rb-2, isolated from a tannery water sample, was identified as Ochrobactrum intermedium, on the basis of 16S rRNA gene sequencing. The influence of factors like temperature of incubation, initial concentration of Cr, mobility of bacteria, and different carbon sources were studied to test the ability of the bacterium to reduce Cr(VI) under variable environmental conditions. The ability of the bacterial strain to reduce hexavalent chromium in artificial and industrial sewage water was evaluated. It was observed that the mechanism of resistance to metal was not due to the change in the permeability barrier of the cell membrane, and the enzyme activity was found to be inductive. Intracellular reduction of Cr(VI) was proven by reductase assay using cell-free extract. Scanning electron microscopy revealed chromium precipitates on bacterial cell surfaces, and transmission electron microscopy showed the outer as well as inner distribution of Cr(VI). This bacterial strain can be useful for Cr(VI) detoxification under a wide range of environmental conditions.

흰목이버섯 대량생산을 위한 용기내 재배 최적화 연구 (Optimization of artificial cultivation of Tremella fuciformis in closed culture bottle)

  • 최성우;장현유;윤정원;이찬
    • 한국버섯학회지
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    • 제6권1호
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    • pp.20-26
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    • 2008
  • 흰목이버섯 균주와 공생균을 수집하고 ITS 5.8S rDNA sequencing을 하여 유전자 서열을 분석하였다. Gene Bank Data homology search 결과 분리된 균의 rDNA 서열이 이 Tremella fuciformis AF042409의 rDNA 서열과 99% 일치하는 것으로 확인되었다. 그리고 함께 분리된 공생균은 같은 방법으로 Annulohhypoxylon stygium 으로 확인하였다. 분리된 T. fuciformis KG 103과 A. stygium KG 201 균주는 PD배지에서 각각 14 mm/14 days과 85 mm/14 days의 균사생육을 나타내었다. T. fuciformis KG 103 균주의 생육최적온도는 $25^{\circ}C$ (14mm/14days) 이었으며, $35^{\circ}C$ 고온과 $15^{\circ}C$이하 저온에서 균사 생장이 억제되었다. A. stygium KG 201은 흰목이버섯균과 유사한 최적온도를 나타내었다. T. fuciformis KG 103 균의 생육 최적 pH는 5.0이었으며, A. stygium KG 201도 pH 5.0에서 생육이 가장 왕성하였다. 흰목이버섯 종균용 최적 배지로 참나무톱밥 77.5%, 미강 20%, 석고 1.5%, 황백당 1%가 선정되었다. T. fuciformis KG 103과 A. stygium KG 201혼합 종균을 제조하고 흰목이버섯 자실체생산을 위한 병속재배 방법을 확립하였다. 콘코브(Corn cob) (77%와 52%)가 사용한 재료 중 최적의 자실체 성장률을 나타냈으며, 콘코브 함량을 줄일수록 생육이 저조하였다. 면실박과 참나무톱밥은 단독 사용시 생육이 저조하였고, 콘코브를 첨가시 수율이 증대되었다. 최적수분농도는 55%로 결정되었다.

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잿빛만가닥버섯(Lyophyllum decastes)의 발효톱밥에 의한 인공재배 특성에 관한 연구 (Development on Artificial Cultivation method of Hatakeshimeji (Lyophyllum decastes) using fermented sawdust substrate)

  • 우성미;박용환;유영복;신평균;장갑열;이강효;성재모
    • 한국버섯학회지
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    • 제7권4호
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    • pp.156-162
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    • 2009
  • 잿빛만가닥버섯의 인공재배법을 개발하고자 국내 외에서 수집한 균주의 균사배양 특성을 검정하고 자실체 발생을 위한 다양한 재배방법을 구명하였다. 잿빛만가닥버섯 9균주를 공시하여 선발한 결과는 다음과 같다. 균사생장속도가 빠르고 균사밀도가 양호한 SPA 202 및 SPA 205균주를 선발하였으며 선발배지는 BC배지가 가장 양호하였다. 인공재배시 참나무톱밥과 포플러톱밥이 균사생장속도가 가장 빠르고 균사밀도가 가장 치밀하였으며 최적배지는 발효톱밥을 사용한 참나무톱밥(Quercus aliena)40%+포플러톱밥(Populus deltoides)40%+미강(Rice bran)10%+밀기울(Wheat bran)10% 배지로서 이 때균사배양적온은 $20-25^{\circ}C$이며 배양기간은 접종 후 50일, 복토 후 7일이 소요되었으며, 원기형성 및 자실체생육기간은 $17-18^{\circ}C$에서 각각 10~11일, 7~8일이 소요되었다. SPA 202균주의 수확적기의 자실체는 갓의 직경이 60mm, 대의 길이가 67mm이며, 수량은1,100ml병당 130g이었으며 SPA 205균주의 수확적기의 자실체는 갓의 직경이 51mm, 대의 길이가 81mm, 129g이었다. 잿빛만가닥버섯 인공재배 결과, 배지는 발효톱밥 처리구가 미발효톱밥 처리구보다 수량이 많아 SPA 202균주와 SPA 205균주가 가장 양호한 균주로 선발되었다.

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