• 제목/요약/키워드: Co-incubation

검색결과 508건 처리시간 0.032초

In Situ Measurement of Diel Periodicity in Urea Decomposition in a Reed Zone of Lake Biwa, Japan

  • Tachibana, Junji;Kondo, Kunio;Seike, Yasushi;Osamu, Mitamura
    • 생태와환경
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    • 제38권spc호
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    • pp.31-38
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    • 2005
  • Diel change in urea decomposition activity of epiphytic algae on Phragmites stems and phytoplankton in a shallow littoral reed zone in the south basin of Lake Biwa was investigated with an in situ technique using $^{14}C$-labelled urea. The daily rates of urea decomposition (sum of urea carbon incorporation rate and $CO_2$ liberation rate) by epiphytic and planktonic algae were calculated as 180 ${\mu}$ mole urea surface shoot area $m^{-2}\;day^{-1}$ and 210 ${\mu}$ mole urea $m^{-3}\;day^{-1}$. The chlorophyll a specific urea decomposition rates of epiphytic and planktonic algae were 4.7 to 6.4 and 4.4 to 6.2 ${\mu}$ mole urea mg chl. $a^{-1}$ incubation $time^{-1}$ in daytime and 4.2 to 5.7 and 2.4 to 3.5 ${\mu}$ mole urea mg chl. $a^{-1}\;time^{-1}$ in nighttime, respectively. High values were obtained during 12:00 ${\sim}$ 18:00 and low values during 00:00 ${\sim}$ 06:00 for both epiphytic and planktonic algal communities. A clear diel periodicity in the urea decomposing activity of the planktonic algae was observed. The activity of the epiphytic algae, on the other hand, showed no destinctive variation during a day. The present results indicate that epiphytic algae are one of the significant urea decomposers in a reed zone, and that the diel patterns are quite difference between both algal communities.

지방기질세포 치료가 당뇨섬유아세포의 증식과 교원질합성에 미치는 영향 (Effect of lipoaspirate cell autograft on proliferation and collagen synthesis of diabetic fibroblasts in vitro)

  • 송선호;한승규;전경욱;김우경
    • Archives of Plastic Surgery
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    • 제36권6호
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    • pp.679-684
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    • 2009
  • Purpose: Human lipoaspirate cells are relatively easy to obtain in large quantities without cell culture. The aim of this in vitro pilot study was to determine the effect of cell therapy using uncultured lipoaspirate cells on cell proliferation and collagen synthesis of diabetic fibroblasts, which are the major contributing factors in wound healing. Methods: In order to get diabetic fibroblasts, dermis tissues were obtained from foot skin of diabetic patients who underwent debridements or toe amputations(n = 4). In order to isolate lipoaspirate cells, the same diabetic patients' abdominal adipose tissues were obtained by liposuction. The diabetic fibroblasts were co - cultured with or without autogenous lipoaspirate cells using porous culture plate insert. Initial numbers of the lipoaspirate cells and diabetic fibroblasts seeded were 15,000 cells/well, respectively. For cell proliferation assay, two treatment groups were included. In group I, diabetic fibroblasts were cultured with the insert having no cells, which serves as a control. In group II, the lipoaspirate cells were added in the culture plate insert. For collagen synthesis assay, one additional group(group III), in which diabetic fibroblasts were not seeded in the well and only lipoaspirate cells inside the insert were incubated without diabetic fibroblasts, was included for a reference. Results: One hundred to one hundred sixty thousand lipoaspirate cells were isolated per ml of aspirated adipose tissue. After 3 - day incubation, the mean cell numbers in group I and II were 17,294/well and 22,163/well. The mean collagen level in group I, II, and III were 29, 41, and 2 ng/ml, respectively. These results imply that both cell proliferation and collagen synthesis in the lipoaspirate cell treatment group were 28 and 44 percents higher than in the control group, respectively(p < 0.05). Conclusion: Uncultured lipoaspirate cell autografts may stimulate the wound healing activity of diabetic fibroblasts.

Polychlorobiphenyl (PCB) 토양오염복원: PCB 제거 토양미생물들의 군집과 기능을 효과적으로 분석하는 신 genomics 방법개발에 관한 연구

  • 박준홍
    • 한국지하수토양환경학회:학술대회논문집
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    • 한국지하수토양환경학회 2005년도 총회 및 춘계학술발표회
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    • pp.28-30
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    • 2005
  • Because of high population diversity in soil microbial communities, it is difficult to accurately assess the capability of biodegradation of toxicant by microbes in soil and sediment. Identifying biodegradative microorganisms is an important step in designing and analyzing soil bioremediation. To remove non-important noise information, it is necessary to selectively enrich genomes of biodegradative microorganisms fromnon-biodegradative populations. For this purpose, a stable isotope probing (SIP) technique was applied in selectively harvesting the genomes of biphenyl-utilizing bacteria from soil microbial communities. Since many biphenyl-using microorganisms are responsible for aerobic PCB degradation In soil and sediments, biphenyl-utilizing bacteria were chosen as the target organisms. In soil microcosms, 13C-biphenyl was added as a selective carbon source for biphenyl users, According to $13C-CO_2$ analysis by GC-MS, 13C-biphenyl mineralization was detected after a 7-day of incubation. The heavy portion of DNA(13C-DNA) was separated from the light portion of DNA (12C-DNA) using equilibrium density gradient ultracentrifuge. Bacterial community structure in the 13C-DNAsample was analyzed by t-RFLP (terminal restriction fragment length polymorphism) method. The t-RFLP result demonstates that the use of SIP efficiently and selectively enriched the genomes of biphenyl degrading bacteria from non-degradative microbes. Furthermore, the bacterial diversity of biphenyl degrading populations was small enough for environmental genomes tools (metagenomics and DNA microarrays) to be used to detect functional (biphenyl degradation) genes from soil microbial communities, which may provide a significant progress in assessing microbial capability of PCB bioremediation in soil and groundwater.

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A Specific Pullulanase for ${\alpha}$-1,6-Glucosidic Linkage of Glucan from Thermus caldophilus

  • Moon-Jo Lee;June-Ki Kim;Kyung-Soo Nam;Jin-Woo Park;Cher-Won Hwang;Dong-Soo Kim;Cheorl-Ho Kim
    • Journal of Life Science
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    • 제9권1호
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    • pp.26-34
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    • 1999
  • A thermostable pullulanase has been isolated and purified from Thermus caldophilus GK-24 to a homogeneity by gel-filtration and ion-exchange chromatography. The specific activity of the purified enzyme was 431-fold increase from the crude culture broth with a recovery of 11.4%. The purified enzyme showed $M_{r}$ of 65 kDa on denaturated and natural conditions. The pI of the enzyme was 6.1 and Schiff staining was negative, suggesting that the enzyme is not a glycoprotein. The enzyme was most active at pH 5.5. The activity was maximal at $75^{\cire}C$ and stable up to $95^{\cire}C$ for 30 min at pH 5.5. The enzyme was stable to incubation from pH 3.5 to pH 8.0 at $4^{\cire}C$ for 24hr. The presence of pullulan protected the enzyme from heat inactivation, the extent depending upon the substrate concentration. The activity of the enzyme was simulated by $Mn^{2+}$ ion, }$Ni^{2+}$, $Ca^{2+}$, $Co^{2+}$ ions. The enzyme hydrolyzed the ${\alpha}$-1,6-linkages of amylopectin, glycogens, ${\alpha}$, ${\beta}$-limited dextrin, and pullulan. The enzyme caused the complete hydrolysis of pullulan to maltotriose and the activity was inhibited by $\alpha$, $\beta$, or $\gamma$-cyclodextrins. The $NH_{2}$-terminal amino acid sequence [(Ala-Pro-Gln-(Asp of Tyr)-Asn-Leu-Leu-Xaa-ILe-Gly-Ala(Ser)] was compared with known sequences of various sources and that was compared with known sequences of various sources and that was different from those of bacterial and plant enzymes, suggesting that the enzymes are structurally different.

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In Vitro Development of Porcine Parthenogenetic Embryos under the Oil-free Culture System

  • Park, Sang-Kyu;Choi, Young-Ju;Roh, Sang-Ho
    • 한국수정란이식학회지
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    • 제25권4호
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    • pp.259-262
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    • 2010
  • Optimization of the preimplantation mammalian embryo culture condition was widely focused on refining medium composition under the name of chemically defined media. However, recent research revealed that the alteration of physical environment can be a crucial factor to a successful embryo development. In this study, under the same embryo density, a novel culture device named oil-free micro tube culture (MTC) system was evaluated using porcine parthenogenetic embryos. The activated oocytes were placed into the 0.2 ml thin-wall flat cap PCR tube and cultured to the blastocyst stage. As a preliminary step, embryo density and culture medium volume were optimized under a standard drop culture system. The optimal embryo density range for in vitro culture was 0.5 embryos per ${\mu}l$ in $20\;{\mu}l$ drop (20.5%) and 1.0 embryos per ${\mu}l$ in $10\;{\mu}l$ drop (20.6%). Based on these results, we compared drop culture system and 'MTC' system in terms of the developmental rate to the blastocyst stage. In $20\;{\mu}l$ medium volume, the 'MTC' system showed similar blastocyst formation rate when compared with drop culture system (20.2% versus 20.5%, respectively) while the 'MTC' system showed lower blastocyst formation rate than drop culture system in $10\;{\mu}l$ one (12.7% versus 20.0%, respectively). Therefore the $20\;{\mu}l$ MTC system may be an alternative incubation system for short-distance embryo transport without carrying the $CO_2$ incubator and this provides novel embryo culture device to clinical veterinary embryologists.

Zoledronate이 UMR-106 세포의 증식과 조골세포 형성에 미치는 영향 (Zoledronate(Zometa(R))inhibits the formation of osteoblast in rat osteoblastic cell line UMR-106)

  • 정기훈;류동목;지유진;이덕원;이현우
    • 대한치과의사협회지
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    • 제46권10호
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    • pp.623-632
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    • 2008
  • Purpose : The purpose of this study is to identify the effect of zoledronate(Zometa(R)), which is most common nitrogen containing bisphosphonate, on survival, proliferation, and differentiation of osteoblast. Material & Methods: Twenty four cell culture plates containing essential medium were seeded with UMR-106 cell lines, at density of 5 x $10^4 cells per plates. Each plates were incubated with 5% $CO^2 incubator at $37^{\circ}C$. Starting from 2 days after incubation, cell culture medias were replaced, and added with osteogenesis induction media and 0, 0.01, 0.1, 0.5, 1, $3\muM$ of zoledronate(Zometa(R)), every 2 days, for 12 days. Control group was plates not added with zoledronate($0\muM$), and experiment group were plates added with different concentration of zoledronates(0, 0.01, 0.1, 0.5, 1, $3\muM$). Mature osteoblasts were identified with Alizarine Red staining, and protein samples were collected. Optical density was determined at wavelength of 405nm with ELISA reader. For viability analysis, cells were harvested and incubated with propidium iodide, and analysed with flow cytometry. Western blot technique was used to analyse Runx2 protein of osteoblast. Results : Secretion of bone matrix decreased as zoledronate concentration increased, and zoledronate did not effect survival rate of UMR-106 cells when measured with flow cytometer. Expression of Runx2 protein was inhibited as zoledronate concentration increased. Conclusion : From the results, we were able to identify that increase of zoledronate concentration inhibited differentiation of UMR-106 cell to osteoblast, without effecting quantity or survival rate.

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시아네이트 에스터 수지의 화학유변학적 거동 및 탄소섬유강화 고분자 복합재료의 물성 (Chemorheological Behavior of Cyanate Ester Resin and Properties of Carbon Fiber Reinforced Polymer Composites)

  • 나효열;윤병철;김승환;이성재
    • Elastomers and Composites
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    • 제48권2호
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    • pp.133-140
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    • 2013
  • 탄소섬유강화 고분자(CFRP) 복합재료는 고분자 매트릭스 내에 탄소섬유를 강화제로 사용한 복합재료이다. 최근 고온 및 고진공 조건이 요구되는 항공우주 및 전자산업용 고성능 재료로 사용하기 위해 높은 열안정성과 낮은 기체방출 특성을 갖는 CFRP 복합재료가 활용되고 있다. 이러한 용도에 시아네이트 에스터 수지는 가장 적합한 매트릭스 수지로 꼽히고 있다. 본 연구에서는 시아네이트 에스터 수지와 촉매의 조합, 경화 거동 및 경화 사이클을 최적화하기 위해 화학유변학적 거동을 분석하였다. 최적 조건은 촉매 100 ppm을 첨가한 수지 조성물을 $150^{\circ}C$에서 경화한 경우로 나타났다. 열안정성과 기체방출 특성을 분석한 결과 경화된 수지 조성물은 열분해 온도 $385^{\circ}C$ 및 전체질량손실 0.29%를 나타내었다. 설정한 수지 조성 및 경화 조건을 사용하여 CFRP 프리프레그 및 이를 적층한 복합재료를 제조하였다. 복합재료의 인장 탄성률을 이론적 모델과 비교한 결과 매우 일관성이 있었다.

고내상 에멀젼 중합에 의한 폴리스티렌/폴리도파민-탄소나노튜브 미세기공 발포체의 제조, 모폴로지 및 전기 전도도 (Preparation, Morphology and Electrical Conductivity of Polystyrene/Polydopamine- Carbon Nanotube Microcellular Foams via High Internal Phase Emulsion Polymerization)

  • 김하승;나효열;이종헌;이성재
    • 폴리머
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    • 제39권2호
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    • pp.293-299
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    • 2015
  • 전기 전도성을 갖는 발포체를 개발하기 위해 고내상 에멀젼(HIPE) 중합법으로 폴리스티렌(PS)/폴리도파민-탄소나노튜브(PDA-CNT) 미세기공 발포체를 제조하여 발포체의 모폴로지 및 전기 전도도를 고찰하였다. 전도성을 부여하기 위한 나노충전제로 CNT를 사용하였는데 수분산성과 HIPE의 안정성을 향상시키기 위하여 CNT 표면을 친수성인 PDA로 개질한 PDA-CNT를 사용하였다. PDA-CNT는 분산성이 우수하여 첨가량을 높여 HIPE를 구성할 수 있었고 전도성이 향상된 발포체를 제조할 수 있었다. 제조한 미세기공 발포체는 기공이 상호 연결된 구조의 모폴로지를 보여 주었다. PDA-CNT의 함량 증가에 따라 HIPE의 항복응력 및 저장 탄성률은 증가하였고 제조된 발포체의 기공 크기는 작아졌다. 전기적 임계점을 보여주는 PDA-CNT 함량은 대략 0.58 wt%였고 PDA-CNT 함량을 5 wt% 첨가했을 때의 전기 전도도는 $10^{-3}S/m$를 나타내었다.

도핵승기탕(桃核承氣湯) 자궁경부암세포(子宮經部癌細胞)(HeLa cell)의 apoptosis에 미치는 영향(影響) (Dohaekseungkitang extract induced apoptosis in Human Cervical carcinoma HeLa cells)

  • 강용구;안규환;공복철;김송백;조한백
    • 대한한방부인과학회지
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    • 제19권2호
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    • pp.77-91
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    • 2006
  • Purpose : To address the ability of Dohaekseungkitang (DST: a commonly used herb formulation in Korea, Japan and China to have anti-cancer effect on cervical carcinoma), we investigated the effects of DST on programmed cell death (apoptosis) in HeLa human cervical carcinoma cells. Methods : We cultured HeLa cell which is human metrocarcinoma cell in D-MEM included 10% fetal bovine serum(Hyclone Laboratories) below $37^{\circ}C$, 5% CO2. Then we observed apoptosis of log phage cell which is changed cultivation liquid 24 Hours periodically. Results : After the treatment of DST for 48 hours, apoptosis occurred in a dose-dependent manner. In this study, we have shown that DST induces calpain and the associated caspase-8 and -9 activations. Apoptosis was prevented by pre-incubation of the cells with the calcium cHeLator-BAPTA-AM, calcium channel blocker-Nif edipine or Ryonidine agonist-Ryonidine peptide, implicating calcium in the apoptotic process. Ubiquitous calpains (mu- and m-calpain) have been repeatedly implicated in apoptosis, especially in calcium-related apoptosis. However this study showed 1hat either calpain inhibitor-calpastin or caspase-3 inhibitor-DEVD- did not blocked the herb formulation-induced apoptosis in HeLa human cervical carcinoma cells. D ST initiates a cell death pathway that is partially dependent of caspases. DST-induced apoptosis requires caspase-independent mechanism. Conclusion : We conclude that DST-induced calpain activation triggers the intrinsic apoptotic pathway in which caspase-independent mechanism is also involved.

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Reticuloendotheliosis virus의 닭에 대한 면역억제효과와 병원성 (Immunosuppressive effects and pathogenicity of a Korean isolate of reticuloendotheliosis virus in chickens)

  • 한명국;김선중
    • 대한수의학회지
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    • 제40권2호
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    • pp.311-323
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    • 2000
  • Immunosuppressive effects of reticuloendotheliosis virus (REV) infection in chickens were investigated. Primary antibody responses to Newcastle disease virus (strain B1) and sheep red blood cells were significantly low in chickens inoculated with the local isolate 89-74 of REV compared to those of uninfected chickens. In chickens infected with REV strain T or 89-74, blastogenesis of spleen cells and peripheral blood lymphocytes (PBL) to concanavalin A (Con A) was severely suppressed. When specific pathogen free (SPF) chickens were inoculated with the isolate, the suppressive effect was observed up to 7 weeks of age while, in the contact infected chickens, the suppression was absent. Similar suppressive effects were observed in chickens inoculated with REV strain T at 2, 3 and 4 weeks of age. When spleen cells or PBL from uninfected chickens were co-cultured with spleen cells or PBL from chickens infected with REV at 1 day-old or 2 week-old, the blastogenesis of the normal cells was suppressed. The suppressive effect of PBL from REV-infected chickens on normal lymphocytes was abrogated by the treatment with trypsin. However the suppressive activity of the REV-infected PBL was not influenced at removing machrophage from the cell suspension by incubation in plastic petri dishes. In addition to the immunosuppression, chickens infected with the REV isolate showed abnormal feather development (nakanuke), anemia, paralysis and retarded growth. Three out of 11 chickens inoculated with the isolate at day-old died between 6 and 9 weeks of age by bacterial infections.

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