• Title/Summary/Keyword: CR2 cells

검색결과 217건 처리시간 0.025초

고정화담체를 이용한 염색폐수의 난분해성 유기물질 처리 (Treatment of Refractory Organics in Dyeing Wastewater by Using Cell Immobilized Pellets)

  • 한덕규;배우근;조영진;원호식;이용우
    • 대한환경공학회지
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    • 제27권9호
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    • pp.917-922
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    • 2005
  • 섬유 염색폐수는 PVA, 유기정련제, 각종 염료 등이 함유되어 있어 처리가 매우 어려운 난분해성 유기폐수로 분류되어 있다. 본 연구에서는 생물학적 처리방법으로 염색폐수의 난분해성 유기물질을 효과적으로 처리할 수 있는 방안으로 활성슬러지를 PEG(Polyethylene glycol)에 포괄고정화한 담체를 고정상 반응기를 이용한 공정에 적용시켜 난분해성 유기물질의 처리효율을 평가하고 적정 운전인자를 검토하였다. 별도로 적응과정을 거치지 않은 활성슬러지를 고정화한 담체를 고정상 반응기에 충전하여 호기성 상태에서 연속식으로 운전하였다. 이때 유입수는 $SCODE_{Cr}$ 약 330 mg/L, $SBOD_5$ 20 mg/L 이하의 생물학적 1차 처리수를 사용하였다. 체류시간의 변화에 따른 각 반응기의 난분해성 유기물질 제거효율을 비교한 결과 EBCT 6 hr을 제외하고는 모두 50% 이상의 유기물($CODE_{Cr}$) 제거효율을 보였고 $COD_{Mn}$의 경우 배출허용기준치 90 mg/L보다 평균 18 mg/L 이상 낮았다. EBCT $6{\sim}24\;hr$ 결과들을 비교한 결과 EBCT 8 hr이 유기물 제거효율과 배출허용기준을 고려할 때 경제적이고 안정된 효율을 얻을 수 있는 적정 체류시간으로 나타났다. 또한 반응기에 충분한 DO를 공급하기 위하여는 주입공기 선속도를 $7\;m^3/m^2{\cdot}hr$ 이상으로 해 주어야 하는 것으로 나타났다. 또한 담체내부 미생물을 동정한 결과 PAH, PVA, Phenol 등 난분해성 물질을 분해하는 균주가 성장하고 있었다. 한편, 고정화담체를 131일에 걸쳐 관찰한 결과 압축강도와 담체내부로의 물질확산에 큰 변화가 없는 것으로 나타났다. 포괄고정화 담체를 이용한 염색폐수처리에서 고정상 반응기는 기존 활성슬러지 공정의 후처리로서 적용 가능할 것으로 판단되어진다.

Evaluation of effect of galvanic corrosion between nickel-chromium metal and titanium on ion release and cell toxicity

  • Lee, Jung-Jin;Song, Kwang-Yeob;Ahn, Seung-Geun;Choi, Jung-Yun;Seo, Jae-Min;Park, Ju-Mi
    • The Journal of Advanced Prosthodontics
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    • 제7권2호
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    • pp.172-177
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    • 2015
  • PURPOSE. The purpose of this study was to evaluate cell toxicity due to ion release caused by galvanic corrosion as a result of contact between base metal and titanium. MATERIALS AND METHODS. It was hypothesized that Nickel (Ni)-Chromium (Cr) alloys with different compositions possess different corrosion resistances when contacted with titanium abutment, and therefore in this study, specimens ($10{\times}10{\times}1.5mm$) were fabricated using commercial pure titanium and 3 different types of Ni-Cr alloys (T3, Tilite, Bella bond plus) commonly used for metal ceramic restorations. The specimens were divided into 6 groups according to the composition of Ni-Cr alloy and contact with titanium. The experimental groups were in direct contact with titanium and the control groups were not. After the samples were immersed in the culture medium - Dulbecco's modified Eagle's medium[DMEM] for 48 hours, the released metal ions were detected using inductively coupled plasma mass spectrometer (ICP-MS) and analyzed by the Kruskal-Wallis and Mann-Whitney test (P<.05). Mouse L-929 fibroblast cells were used for cell toxicity evaluation. The cell toxicity of specimens was measured by the 3-{4,5-dimethylthiazol-2yl}-2,5-diphenyltetrazolium bromide (MTT) test. Results of MTT assay were statistically analyzed by the two-way ANOVA test (P<.05). Post-hoc multiple comparisons were conducted using Tukey's tests. RESULTS. The amount of metal ions released by galvanic corrosion due to contact between the base metal alloy and titanium was increased in all of the specimens. In the cytotoxicity test, the two-way ANOVA showed a significant effect of the alloy type and galvanic corrosion for cytotoxicity (P<.001). The relative cell growth rate (RGR) was decreased further on the groups in contact with titanium (P<.05). CONCLUSION. The release of metal ions was increased by galvanic corrosion due to contact between base metal and titanium, and it can cause adverse effects on the tissue around the implant by inducing cytotoxicity.

외래유전자를 주입한 소 수정란에서 형질전환가능 수정란의 선발 (Preselection of Bovine Blastocysts Expressing Exogeneous Gene Following Microinjection)

  • 공일근
    • 한국가축번식학회지
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    • 제21권2호
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    • pp.167-176
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    • 1997
  • 본 연구는 수정란을 수란우에 이식하기 이전에 형질전환가능 수정란을 선발할 수 있다면 형질전환동물의 생산에 크게 도움이 되므로 3.2 kb $\beta$-actin promoter (lacZ/n대) DNA를 미세주입하여 배반포기배에서의 발현을 확인하여 이들을 선발할 수 있는가를 규명하고자 하엿다. 채란된 난포란은 10%FBS, 5$\mu\textrm{g}$/ml LH, 0.5$\mu\textrm{g}$/ml FSH, 100 unit/ml penicillin 및 100 $\mu\textrm{g}$/ml streptomycin이 함유된 TCM199에 22~24 시간동안 체외성숙을 유도후 5$\mu\textrm{g}$/ml heparin으로 수정능획득을 유도한 1$\times$106 sperm/ml의 정자로 체외수정을 시켰다. 체외수정후 18~20시간째에 vortexing에 의해 과립막세포를 제거하고 원심분리시켜 자/웅전핵이 확인되는 수정란의 핵에 3~4 ng/${\mu}\ell$ lacZ/neo DNA를 미세주입하였다. 모든 수정란의 배양은 3 mg/ml BSA, 20${\mu}\ell$/ml NEM AMINO acids 및 40${\mu}\ell$/ml BME amino acids가 함유되어 있는 CR1aa 배양액에 neo/DNA로 transfected 된 BRL 단층에서 실시하였다. G418에 대한 적정농도를 찾기 위하여 정상적인 수정란에 0, 50, 100 및 200 $\mu\textrm{g}$/ml G418를 첨가하여 배양한 결과 8일째에 30.3%(44/145), 8.7%(13/150), 0.7%(1/151) 및 0% (0/134)의 수정란이 배반포기까지 발달하였다. 그래서 본 실험에서는 일정하게 100$\mu\textrm{g}$/ml G418을 첨가하여 배양하였다. 총 1,127개의 수정란을 미세주입후 G418 없는 배양액에서 710개 (63.0%)가 분할하였다. 미세주입후 48시간째에 2-세포기이상 분할된 수정란을 대조구 및 100$\mu\textrm{g}$/ml G418처리구를 무작위로 할당하여 배양하였으며, 또한 740개의 정상수정란도 같은 반복수로 배양을 실시하였다. 미세주입한 수정란은 8일 후 11.6%(26/255) 및 5.2% (14/267)가 대조구 및 G418 처리구에서 배반포기까지 발달하였으며 정상수정란은 27.2% (151/740)가 배반포기 배까지 발달하였다. 미세주입후 대조구에서는 23.1$\pm$2.6/70.7$\pm$4.7 (32.7%)의 할구가 $\beta$-Gal 활력을 보였고, 반면에 100$\mu\textrm{g}$/ml G418 처리구에서는 40.3$\pm$4.1/48.8$\pm$7.5 (82.6%)가 $\beta$-Gal 활력을 보였다. 비록 mosaic 형태로 외래유전자가 발현되었지만 대조구에서 87.0% (26/30개) 배반포기가 $\beta$-Gal 활력을 보인 반면, G418 처리구에서는 모든 배반포기가 $\beta$-Gal 활력을 보였다 (P<0.05). 그러나 대조구 및 G418 처리구의 ICM colony에서는 영양배엽과 내배엽을 제외한 epiblast에서는 확인되지 않았다. 그러나 이 결과로부터 $\beta$-actin promoter/lacZ gene이 integration되지 않는 것인지 또는 다만 염색 확인이 되지 않는 것인지를 판단할 수는 없다. 이상의 결과는 미세주입후 G418에서 배양한 배반포기배에서는 대부분의 할구에서 주입된 gene을 발현하고 있었으나 ICM colony에서는 특히 epiblast에서는 발현되지 않거나 침묵하고 있었다. 비록 G418 처리구에서 훨씬 더 높은 비율로 주입된 gene이 발현되고 있으나 총세포수는 유의적으로 감소하여 이후 형질전환동물의 생산과 ES like-cell의 설립에는 감소될 것으로 사려된다. 그러나 형질전환 수정란의 선발 및 형질전환동물의 생산능력에 관해서는 더 많은 연구가 필요하다고 사려된다.

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Efficient aerobic denitrification in the treatment of leather industry wastewater containing high nitrogen concentration

  • Kang, Kyeong Hwan;Lee, Geon;Kim, Joong Kyun
    • Environmental Engineering Research
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    • 제20권1호
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    • pp.79-87
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    • 2015
  • To treat leather industry wastewater (LIW) containing high nitrogen concentration, eight aerobic denitrifiers were isolated from sludge existing in an LIW-treatment aeration tank. Among them, one strain named as KH8 had showed the great ability in denitrification under an aerobic condition, and it was identified as Pseudomonas aeruginosa R12. The aerobic denitrification ability of the strain KH8 was almost comparable to its anaerobic denitrification ability. In lab-scale aerobic denitrifications performed in 1-L five-neck flasks for 48 hr, denitrification efficiency was found to be much improved as the strain KH8 held a great majority in the seeded cells. From the nitrogen balance at the cell-combination ratio of 10:1 (the strain KH8 to the other seven isolates) within the seeded cells, the percentage of nitrogen loss during the aerobic denitrification process was estimated to be 58.4, which was presumed to be converted to $N_2$ gas. When these seeded cells with lactose were applied to plant-scale aeration tank for 56 day to treat high-strength nitrogen in LIW, the removal efficiencies of $COD_{Cr}$ and TN were achieved to be 97.0% and 89.8%, respectively. Under this treatment, the final water quality of the effluent leaving the treatment plant was good enough to meet the water-quality standards. Consequently, the isolated aerobic denitrifiers could be suitable for the additional requirement of nitrogen removal in a limited aeration-tank capacity. To the best of our knowledge, this is the first report of aerobic denitrifiers applied to plant-scale LIW treatment.

Harnessing CRISPR-Cas adaptation for RNA recording and beyond

  • Gyeong-Seok Oh;Seongjin An;Sungchul Kim
    • BMB Reports
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    • 제57권1호
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    • pp.40-49
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    • 2024
  • Prokaryotes encode clustered regularly interspaced short palindromic repeat (CRISPR) arrays and CRISPR-associated (Cas) genes as an adaptive immune machinery. CRISPR-Cas systems effectively protect hosts from the invasion of foreign enemies, such as bacteriophages and plasmids. During a process called 'adaptation', non-self-nucleic acid fragments are acquired as spacers between repeats in the host CRISPR array, to establish immunological memory. The highly conserved Cas1-Cas2 complexes function as molecular recorders to integrate spacers in a time course manner, which can subsequently be expressed as crRNAs complexed with Cas effector proteins for the RNA-guided interference pathways. In some of the RNA-targeting type III systems, Cas1 proteins are fused with reverse transcriptase (RT), indicating that RT-Cas1-Cas2 complexes can acquire RNA transcripts for spacer acquisition. In this review, we summarize current studies that focus on the molecular structure and function of the RT-fused Cas1-Cas2 integrase, and its potential applications as a directional RNA-recording tool in cells. Furthermore, we highlight outstanding questions for RT-Cas1-Cas2 studies and future directions for RNA-recording CRISPR technologies.

참다래의 이화학적 성분, 항균 활성 및 폐암 세포 증식 억제 효과 (Chemical Composition of Kiwifruits, Their Anti-microbial Activity and Their Hyperplasia Inhibition Effect of against Lung Cancer Cells)

  • 박용서;이건순;코삭;박윤점;오대민;허북구
    • 동아시아식생활학회지
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    • 제19권2호
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    • pp.202-209
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    • 2009
  • This study was conducted to gather the basic data on the spread of the domestic kiwifruits, and the development of the manufactured goods and the health functional foods produced using kiwifruits. We determined the chemical compositions of four types of kiwifruits cultivated in Korea, Daeheung, Bidan, Haegeum and Hayward. In addition, we measured the anti-microbial activities and cytotoxicities of these types of kiwifruits. The vitamin C contents of the kiwifruits increased in the order of Bidan (93.82 mg/100 g), Daeheung (85.89 mg/100 g), Haegeum (83.73 mg/100 g) and Hayward (75.28 mg/100 g). The total amino acids contents per 100 g of kiwifruit (dry weight basis) were 483.97 mg (Haegeum), 453.08 mg (Hayward), 437.27 mg (Bidan) and 369.35 mg (Daeheung). The K and Ca contents of the kiwifruits ranged from 14.56~37.12 mg/L and 1.94~8.24 mg/L, respectively; however, the Fe, Mg, Zn and Cr contents all less than 1.83 mg/L. The antimicrobial activities of methanol extracts of kiwifruits against five gram positive bacteria at concentration of 2,000 mg/L in terms of inhibition diameter ranged from 8.8~12.8mm, while raged from 9.2~13.1mm against three gram negative strains of bacteria. The hyperplasia inhibition of lung cancer cells (Calus-6) by 800 mg/L kiwifruits extracts of Bidan, Haegeum, Daeheung and Hayward kiwifruits were 21.2%, 9.5%, 6.7% and 5.0%, respectively. Consequently, it was assumed that kiwifruits was rich in vitamin C, amino acids and K, and that they would therefore be useful in processed goods.

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Chemosensitizing Activity Against Human Leukemia Cell of Crude Extracts of Native Camellia(Camellia japonica) in Jeonnam

  • Hwang, Eun-Ju;Lee, Sook-Young
    • 한국응용약물학회:학술대회논문집
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    • 한국응용약물학회 2003년도 Annual Meeting of KSAP : International Symposium on Pharmaceutical and Biomedical Sciences on Obesity
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    • pp.85-85
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    • 2003
  • This study has been undertaken to increase availability of native camellia in Jeonnam as a medicinal resource and to isolate the effective components from them. Multidrug resistance(MDR) by tumor cells is a major obstacle to successful cancer chemotherapy. We report that the crude extracts of camellia flowers, leaves has a chemosensitizing effect that can reverse Pgp-mediated MDR by increasing the intracellular accumulation of drugs. The cytotoxic and chemosensitizing effects of MeOH extract from 12 spp. citrus fruits on the AML-2/D100 were determined using MTT assay. Chemosensitizing effects was screened in the presence of vincristine, a good substrate of Pgp. IC$\sub$50/ for extracts in AML-2/WT was found to be 65∼350$\mu\textrm{g}$/$m\ell$ whereas the range of its mean IC$\sub$50/ value in Pgp-overexpressing cells (AML-2/Dl00) in the presence of vincristine was 90∼400$\mu\textrm{g}$/$m\ell$. Of the extracts tested, mature leaf extract displayed the most potent chemosensitizing effect[IC$\sub$50/;100 $\mu\textrm{g}$/$m\ell$, CR;1.06, RF;2.97 in the presence of VCR]. This indicates that the toxicity (IC$\sub$50/;288.89$\mu\textrm{g}$/$m\ell$) of mature leaf extract is minimal at concentrations required for a complete reversal of the drug resistance. Also, this result indicates that crude extracts of camellia mature leaves would contain some principles which have chemosensitizing activity.

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Long Non-coding RNAs are Differentially Expressed in Hepatocellular Carcinoma Cell Lines with Differing Metastatic Potential

  • Fang, Ting-Ting;Sun, Xiao-Jing;Chen, Jie;Zhao, Yan;Sun, Rui-Xia;Ren, Ning;Liu, Bin-Bin
    • Asian Pacific Journal of Cancer Prevention
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    • 제15권23호
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    • pp.10513-10524
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    • 2015
  • Background: Metastasis is a major reason for poor prognosis in patients with cancer, including hepatocellular carcinoma (HCC). A salient feature is the ability of cancer cells to colonize different organs. Long non-coding RNAs (lncRNAs) play important roles in numerous cellular processes, including metastasis. Materials and Methods: In this study, the lncRNA expression profiles of two HCC cell lines, one with high potential for metastasis to the lung (HCCLM3) and the other to lymph nodes (HCCLYM-H2) were assessed using the Arraystar Human LncRNA Array v2.0, which contains 33,045 lncRNAs and 30,215 mRNAs. Coding-non-coding gene co-expression (CNC) networks were constructed and gene set enrichment analysis (GSEA) was performed to identify lncRNAs with potential functions in organ-specific metastasis. Levels of two representative lncRNAs and one representative mRNA, RP5-1014O16.1, lincRNA-TSPAN8 and TSPAN8, were further detected in HCC cell lines with differing metastasis potential by qRT-PCR. Results: Using microarray data, we identified 1,482 lncRNAs and 1,629 mRNAs that were differentially expressed (${\geq}1.5$ fold-change) between the two HCC cell lines. The most upregulated lncRNAs in H2 were RP11-672F9.1, RP5-1014O16.1, and RP11-501G6.1, while the most downregulated ones were lincRNA-TSPAN8, lincRNA-CALCA, C14orf132, NCRNA00173, and CR613944. The most upregulated mRNAs in H2 were C15orf48, PSG2, and PSG8, while the most downregulated ones were CALCB, CD81, CD24, TSPAN8, and SOST. Among them, lincRNA-TSPAN8 and TSPAN8 were found highly expressed in high lung metastatic potential HCC cells, while lowly expressed in no or low lung metastatic potential HCC cells. RP5-1014O16.1 was highly expressed in high lymphatic metastatic potential HCC cell lines, while lowly expressed in no lymphatic metastatic potential HCC cell lines. Conclusions: We provide the first detailed description of lncRNA expression profiles related to organ-specific metastasis in HCC. We demonstrated that a large number of lncRNAs may play important roles in driving HCC cells to metastasize to different sites; these lncRNAs may provide novel molecular biomarkers and offer a new basis for combating metastasis in HCC cases.

Genetic Analysis of Ancient Bones of Cervidae Animals from Archaeological Site in Jeju, Korea

  • Kang, Min-Chul;Han, Sang-Hyun;Jung, Yong-Hwan;Oh, Ju-Hyung;Kim, Gi-Ok;Ko, Jae-Woen;Oh, Moon-You
    • Animal cells and systems
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    • 제11권2호
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    • pp.147-153
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    • 2007
  • DNA extracted from ancient bones of Cervidae animals was examined to identify the species and to determine the phylogenetic relationships to those from extant cervids. Abundant ancient bones were excavated from Kumsung archaeological site in Jeju Island, Korea, and were identified as Cervidae animals based on morphological features of their antlers and lower mandibles. Their mitochondrial DNA (mtDNA) control region (CR) was partially sequenced and subsequently compared with those previously reported in database. The results confirmed that the ancient sequences are lineage of Cervidae. On the phylogenetic trees constructed using the sequence diversity of the CR sequences of family Cervidae, the ancient DNA sequences were found on distinct clusters. The ancient sequences were located in the subfamily Capreolinae cluster, and six ancient sequences were closely related to those of extant Korean roe deer in Jeju Island and Korean Peninsula. Consequently, the results of this study suggest that the roe deer inhabited Jeju Island in ancient times. However, there is no evidence for the existence of subfamily Cervinae, including Sika deer, while it has been described in several historical records. The results suggest that this finding could contribute to understanding of the origin and phylogenetic relationships of extant and ancient roe deer on Jeju Island.

한우 체세포를 이용한 핵이식에서 전기융합 방법이 융합율 및 배발달율에 미치는 영향 (Effect of Electric fusion Methods on Cell Fusion Rate and Embryo Development by Somatic Cell Nuclear Transfer in Korean Native Cattle(KNC))

  • 김은국;김정욱
    • 한국수정란이식학회지
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    • 제18권3호
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    • pp.171-178
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    • 2003
  • 복제수정란 생산에 있어서 수핵란 내 체세포 주입 후 전기적인 융합은 필수과정인데, 이 과정을 거치는 동안 많은 수의 체세포 주입 난자가 융합에 실패하거나 lysis가 일어나게 된다. 본 실험에서는 한우 체세포를 이용하여 핵이식을 실시한 후 수핵세포질과 응합을 시도할 때 전기융합 방법에 따른 융합율과 배발달율을 검토하고자 실시하였다. 공여세포는 한우 귀 세포조직을 채취하여 0.05% trypsin과 EDTA가 첨가된 D-PBS로 세포를 분리한 후 DMEM 배양액으로 계대배양을 실시하여 사용하였다. 핵이식을 위하여 체외성숙시킨 난자를 탈핵용 micro pipette을 이용하여 투명대를 절개하고 수핵 난자의 극체 와 핵을 제거한 후 공여세포를 주입하였으며, 핵이식 수정란은 직접, 간접적인 전기적 자극으로 융합을 실시한 후 calcium iono-phore와 6-DMAP를 이용하여 활성화를 유도하였다. 활성화된 수정란은 38.5$^{\circ}C$, 90% $N_2$, 5% $CO_2$로 조정된 배양기에서 처음 3일은 0.3% BSA가 첨가된 CR1-aa 배양액에서, 배양 4일째부터는 10% FBS가 첨가된 CR1-aa 배양액에서 배양을 실시하였다. 본 연구결과를 요약하면 다음과 같다. 전기자극 융합방법에 따른 수핵난자의 융합율과 lysis율은 electric chamber를 이용하여 간접융합을 실시하였을 경우 51.6%와 10.7%를 보였고, needle을 이용하여 직접 융합을 실시하였을 경우 68.9%의 융합율과 11.5%의 lysis율을 보임으로써 needle을 이용한 직접융합 방법이 유의적으로 높은 융합율을 보였다(P<0.05). 융합 후 체외 발달과정을 살펴보면 난할율에 있어서 needle을 이용했을 시 80.3%로 chamber를 이용했을 시 73.2%보다 유의적으로 높은 난할율을 보였다. 초기배 발달단계인 2∼4세포기의 발달율 역시 needle을 사용한 구가 61.1%로 chamber를 사용한 구 54.1%보다 유의적으로 높은 차이를 보였다. 상실배 단계는 chamber를 사용한 구가 6.7%로 needle을 사용한 구 6.2%보다 약간 높았지만 유의적인 차이는 없었다. 하지만 이식 가능한 단계인 배반포배 발달율에 있어서는 needle을 사용하여 융합을 시도한 구가 26.3%로 chamber를 사용한 구 18.4%보다 유의적으로 높은 발달율을 보였다(P<0.05). electric chamber를 이용하여 전기융합을 시도시전류가 흐르는 wire와 주입된 체세포가 직각을 이루도록 정렬을 시키느라 많은 시간이 소요되고, 주입한 체세포가 세포질과 떨어진 난자는 전기자극을 주어도 융합이 일어날 수 없으며, 높은 전압을 사용하기 때문에 융합된 복제란의 lysis가 많이 발생하는게 가장 큰 단점으로 꼽을 수 있다. 하지만 미세조작기와 needle 방법을 이용하면 낮은 전압을 이용하여 융합을 시도하기 때문에 복제란의 lysis를 줄일 수 있고, 전극과 체세포 주입란을 정렬시키는 과정이 생략되어 시간이 절약되며, 결정적으로 주입된 체세포가 세포질과 떨어져 있더라도 미세조작기로 약간의 압력을 가하여 전기자극을 가할 수 있어서 보다 높은 융합율과 배반포배 발달율을 얻을 수 있기 때문에 needle을 이용한 직접적인 전기융합 방법이 체세포 복제수정란 생산에 효과적이라고 사료된다.