• 제목/요약/키워드: double-counting

검색결과 48건 처리시간 0.022초

Plumbagin from Plumbago Zeylanica L Induces Apoptosis in Human Non-small Cell Lung Cancer Cell Lines through NF-κB Inactivation

  • Xu, Tong-Peng;Shen, Hua;Liu, Ling-Xiang;Shu, Yong-Qian
    • Asian Pacific Journal of Cancer Prevention
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    • 제14권4호
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    • pp.2325-2331
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    • 2013
  • Objective: To detect effects of plumbagin on proliferation and apoptosis in non-small cell lung cancer cell lines, and investigate the underlying mechanisms. Materials and Methods: Human non-small cell lung cancer cell lines A549, H292 and H460 were treated with various concentrations of plumbagin. Cell proliferation rates was determined using both cell counting kit-8 (CCK-8) and clonogenic assays. Apoptosis was detected by annexin V/propidium iodide double-labeled flow cytometry and TUNEL assay. The levels of reactive oxygen species (ROS) were detected by flow cytometry. Activity of NF-${\kappa}B$ was examined by electrophoretic mobility shift assay (EMSA) and luciferase reporter assay. Western blotting was used to assess the expression of both NF-${\kappa}B$ regulated apoptotic-related gene and activation of p65 and $I{\kappa}B{\kappa}$. Results: Plumbagin dose-dependently inhibited proliferation of the lung cancer cells. The IC50 values of plumbagin in A549, H292, and H460 cells were 10.3 ${\mu}mol/L$, 7.3 ${\mu}mol/L$, and 6.1 ${\mu}mol/L$ for 12 hours, respectively. The compound concentration-dependently induced apoptosis of the three cell lines. Treatment with plumbagin increased the intracellular level of ROS, and inhibited the activation of NK-${\kappa}B$. In addition to inhibition of NF-${\kappa}B$/p65 nuclear translocation, the compound also suppressed the degradation of $I{\kappa}B{\kappa}$. ROS scavenger NAC highly reversed the effect of plumbagin on apoptosis and inactivation of NK-${\kappa}B$ in H460 cell line. Treatment with plumbagin also increased the activity of caspase-9 and caspase-3, downregulated the expression of Bcl-2, upregulated the expression of Bax, Bak, and CytC. Conclusions: Plumbagin inhibits cell growth and induces apoptosis in human lung cancer cells through an NF-${\kappa}B$-regulated mitochondrial-mediated pathway, involving activation of ROS.

광역지자체의 산림지 온실가스 인벤토리 시범 산정 (Estimation of the Greenhouse Gas Inventory on Forest Land at Provincial Level)

  • 김경남;이선정;김래현;손영모
    • 한국농림기상학회지
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    • 제16권4호
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    • pp.336-342
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    • 2014
  • 본 연구는 우리나라의 광역지자체의 산림지 온실가스 배출량을 시범산정하기 위해 수행되었다. 온실가스 배출량은 국가 온실가스 인벤토리 산정 지침에 따라 산정하였으며, 이를 위해 임업통계연보 및 산림관련 활동 자료 및 배출계수를 이용하였다. 그 결과 산림지의 2011년 총 배출량은 -58,711Gg $CO_2eq.$, 벌채, 연료재 수집, 산불에 의한 배출량은 19,896Gg $CO_2eq.$로 나타났다. 2011년을 기준으로 산림지에서는 온실가스를 흡수하는 것으로 나타났고, 순 배출량은 -38,815Gg $CO_2eq.$로 나타났다. 광역지자체 별로는 강원도에서 온실가스를 가장 많이 흡수하는 것으로 나타났다. 본 연구의 산정결과는 현재 중복산정을 최대한 배제하고자 많은 통계를 활용하지 않은 것으로, 향후 활동자료 및 배출계수 등을 보완하여 산정할 필요가 있을 것으로 사료된다.

교육 대상에 따른 야외 지질학 탐구 요소 특성 비교 : 지질 답사 교육 사례를 중심으로 (Comparing Elements of Inquiry in Field Geology by Learner Groups: Focusing on Cases of Geologic Fieldwork Education)

  • 정찬미;신동희
    • 대한지구과학교육학회지
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    • 제10권3호
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    • pp.235-253
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    • 2017
  • 본 연구의 목적은 최근 20년간 국내외 문헌에 나타난 지질 답사 교육 사례가 야외 지질학 탐구 요소를 어떻게 반영하고 있는지 교육 대상별로 비교 분석하는 것이다. 분석 대상 사례는 총 53건이며, 분석은 야외 지질학 탐구 요소 및 하위 요소에 대해 수행되었다. 분석 결과, 야외 지질학 탐구 요소 반영양상에서 초중고생 대상 지질 답사 교육 사례와 지질학 관련 전공생 대상 사례 간 명확한 차이가 드러났다. 초중고생 대상 답사는 주로 교육 과정에 기반하여 설계되었기 때문에 야외 지질학 탐구 요소 중 관찰, 표상, 귀추적 추론 중 2~3가지를 포함하는 경우가 많았다. 반면 지질학 관련 전공생 대상 답사는 지질 전문가 양성을 목적으로 하기 때문에 야외 지질학 탐구 요소 중 4~5개를 다양하게 포함하고 있으며 공간적 사고 및 통시적 사고의 비율도 높게 나타났고, 지질도 작성 등 전문 기술을 요구하는 활동이 자주 활용되었다. 과학 교사 대상 답사는 초중고생 및 지질학 관련 전공생 대상 답사의 특징이 혼합된 형태로 나타났다. 포함된 탐구 요소의 다양성, 지질학자가 수행하는 활동과의 유사성, 학습자의 자율성 등을 종합적으로 고려했을 때 지질학 관련 전공생 대상 답사가 다른 교육 대상의 답사보다 야외 지질학 탐구의 실제성이 높은 것으로 평가되었다. 이 연구의 결과를 바탕으로 참 과학 탐구로서 지질 답사교육의 활용을 위한 시사점을 제시했다.

Comparison of Parallel and Fan-Beam Monochromatic X-Ray CT Using Synchrotron Radiation

  • Toyofuku, Fukai;Tokumori, Kenji;Kanda, Shigenobu;Ohki, Masafumi;Higashida, Yoshiharu;Hyodo, Kazuyuki;Ando, Masami;Uyama, Chikao
    • 한국의학물리학회:학술대회논문집
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    • 한국의학물리학회 2002년도 Proceedings
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    • pp.407-410
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    • 2002
  • Monochromatic x-ray CT has several advantages over conventional CT, which utilizes bremsstrahlung white x-rays from an x-ray tube. There are several methods to produce such monochromatic x-rays. The most popular one is crystal diffraction monochromatization, which has been commonly used because of the fact that the energy spread is very narrow and the energy can be changed continuously. The alternative method is the use of fluorescent x-ray, which has several advantages such as large beam size and fast energy change. We have developed a parallel-beam and a fan-beam monochromatic x-ray CT, and compared some characteristics such as accuracy of CT numbers between those systems. The fan beam monochromatic x-rays were generated by irradiating target materials by incident white x-rays from a bending magnet beam line NE5 in 6.5 GeV Accumulation Ring at Tukuba. The parallel beam monochromatic x-rays were generated by using a silicon double crystal monochromator at the bending magnet beam line BL-20BM in Spring-8. A Cadmium telluride (CdTe) 256 channel array detector with 512mm sensitive width capable of operating at room temperature was used in the photon counting mode. A cylindrical phantom containing eight concentrations of gadolinium was used for the fan beam monochromatic x-ray CT system, while a phantom containing acetone, ethanol, acrylic and water was used for the parallel monochromatic x-ray CT system. The linear attenuation coefficients obtained from CT numbers of those monochromatic x-ray CT images were compared with theoretical values. They showed a good agreement within 3%. It was found that the quantitative measurement can be possible by using the fan beam monochromatic x-ray CT system as well as a parallel beam monochromatic X-ray CT system.

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Droplet Digital PCR을 이용한 Bacillus cereus, Staphylococcus aureus, Salmonella Typhimurium과 Escherichia coli O157:H7의 검출 및 정량 (Detection and quantitation of Bacillus cereus, Staphylococcus aureus, Salmonella Typhimurium and Escherichia coli O157:H7 by droplet digital PCR)

  • 김진희;윤진선;이다영;김동호;오세욱
    • 한국식품과학회지
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    • 제48권5호
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    • pp.454-460
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    • 2016
  • 본 논문에서는 E. coli O157:H7, S. Typhimurium, S. aureus와 B. cereus에 대한 ddPCR의 검출 효율과 검출한계를 측정하였으며 동시 검출을 위한 multiplex 검출 가능성을 타진하였다. ddPCR은 PCR mix를 포함한 시료를 15,000-20,000개의 droplet으로 분할하여 PCR 하는 방법으로 droplet reader를 이용하여 droplet의 형광 신호를 계수하였다. 식중독 세균의 표적 DNA를 검출하기 위해 2가지 색의 형광 probe (TaqMan)를 제작하였다. ddPCR은 표적 유전자의 형광 신호를 $100fg/{\mu}L$부터 $10ng/{\mu}L$까지의 DNA를 검출 할 수 있었다. 이후에 두 종류의 식중독 세균에서 프라이머 농도를 달리하여 표적 DNA 증폭 크기의 분포가 서로 다르게 구별할 수 있음을 확인하여 multiplex PCR의 가능성이 있음을 알 수 있었다. ddPCR은 비교적 낮은 검출한계를 가지기 때문에 식품에 적은 농도로 존재하는 식중독 세균의 검출에 활용이 가능할 것으로 생각되었다. 또한 식품의 전처리 조건 확립과 반응조건 확립을 통하여 향후 복잡한 matrix effect를 가지는 식품에서 극 미량의 균 검출도 가능할 것으로 생각되었다.

ssc-miR-185 targets cell division cycle 42 and promotes the proliferation of intestinal porcine epithelial cell

  • Wang, Wei;Wang, Pengfei;Xie, Kaihui;Luo, Ruirui;Gao, Xiaoli;Yan, Zunqiang;Huang, Xiaoyu;Yang, Qiaoli;Gun, Shuangbao
    • Animal Bioscience
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    • 제34권5호
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    • pp.801-810
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    • 2021
  • Objective: microRNAs (miRNAs) can play a role in a variety of physiological and pathological processes, and their role is achieved by regulating the expression of target genes. Our previous high-throughput sequencing found that ssc-miR-185 plays an important regulatory role in piglet diarrhea, but its specific target genes and functions in intestinal porcine epithelial cell (IPEC-J2) are still unclear. We intended to verify the target relationship between porcine miR-185 and cell division cycle 42 (CDC42) gene in IPEC-J2 and to explore the effect of miR-185 on the proliferation of IPEC-J2 cells. Methods: The TargetScan, miRDB, and miRanda software were used to predict the target genes of porcine miR-185, and CDC42 was selected as a candidate target gene. The CDC42-3' UTR-wild type (WT) and CDC42-3'UTR-mutant type (MUT) segments were successfully cloned into pmirGLO luciferase vector, and the luciferase activity was detected after co-transfection with miR-185 mimics and pmirGLO-CDC42-3'UTR. The expression level of CDC42 was analyzed using quantitative polymerase chain reaction and Western blot. The proliferation of IPEC-J2 was detected using cell counting kit-8 (CCK-8), methylthiazolyldiphenyl-tetrazolium bromide (MTT), and 5-ethynyl-2'-deoxyuridine (EdU) assays. Results: Double enzyme digestion and sequencing confirmed that CDC42-3'UTR-WT and CDC42-3'UTR-MUT were successfully cloned into pmirGLO luciferase reporter vector, and the luciferase activity was significantly reduced after co-transfection with miR-185 mimics and CDC42-3'UTR-WT. Further we found that the mRNA and protein expression level of CDC42 were down-regulated after transfection with miR-185 mimics, while the opposite trend was observed after transfection with miR-185 inhibitor (p<0.01). In addition, the CCK-8, MTT, and EdU results demonstrated that miR-185 promotes IPEC-J2 cells proliferation by targeting CDC42. Conclusion: These findings indicate that porcine miR-185 can directly target CDC42 and promote the proliferation of IPEC-J2 cells. However, the detailed regulatory mechanism of miR-185/CDC42 axis in piglets' resistance to diarrhea is yet to be elucidated in further investigation.

남강유역 간흡충감염의 역학적 조사연구 (Epidemiological Studios on Clonorchis sinensis Infection along the Nam-river in Gyeongnam Province, Korea)

  • 배경훈;안영겸
    • Parasites, Hosts and Diseases
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    • 제21권2호
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    • pp.116-186
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    • 1983
  • An epidemiological study on Clonorchis sinensis infection along the Nam-river (total length ; 186km) flowing in Gyeongsang-nam-do, southern part of Korea, was carried out. Formalin-ether concentration technique and Stoll's egg counting method were employed to figure out the prevalence of C. sinensis infection. For the detection of cercariae from Parafossarulus sp., the snail host of C. sinensis, each snail was placed in aerated tap water, and examined for expelled cercariae. For observing the metacercariae the fresh water fish favorably eaten in raw conditions wore prepared by means of pressing the muscles between two slide glasses and/or by digesting them with artificial gastric juice. The fresh water fish were fed to the rabbits to get the worms and to identify the morphology of adult C. sinensis. The results are summarized as follows: 1. Overall Prevalence of C. sinensis infection was 38.75 from 5,291 examinees; 44.15 (1,408 out of 3,196) in male and 30.4% (637 out of 2,095) in female. 2. The prevalence rates were 42.0% at the upper stream, 41.2% around the vicinity of Jinyanglake, 34.2% at Jinju city, 34.2% at middle stream and 40.3% at down stream regions, respectively. 3. By age, the highest positive rate (53.4∼54.3%) was observed in 30 to 59 years of age. In this age group, the rate in males was 59.7∼62.2%, and in females 42.2∼44.4%, In the age group of less than 19 years it was 7.5∼20.9%. 4. By social strata, the positive rate was 16.5% in the primary school children, 22.6% in school students, 46.2% in teachers and local officers and 49.6% in the general inhabitants. 5. The quantitative examinations with the stool collected from clonorchiasis cases revealed that the light infection (less than 4,000/EPG) was 53.6%, moderate infection (4,001∼10,000 /EPG) 30.3% and heavy infection (more than 10,001/EPG) 16.1%, respectively. More than half of total cases examined were light infection, and 73.2% of female examinees were lightly infected with this cuke. 6. The average value of EPG was 4,963 (male, 6,057; female, 2,557 and the highest value was obtained from the age group of 30 to 59 years (5,240∼6,454). 7. The prevalence of Metagonimus yokogawai infection in humans was 5.5%, and 89.8% of metagonimiasis cases wore double-infected arith C. sinensis. The highest prevalence rate was observed in Jinju city (11.2%). 8. Total of 5,005 Parafossarulus sp., the snail intermediate host of C. sinensis, were examined for the detection of cercariae. The cercarial expulsion rate was 0.34%, and the snails collected in Jin-yang-lake side and in the down stream expelled mainly the cercariae of C, sinensis. 9. About 788 cercariaejday (range: 127∼1,503) were expelled daily from a snail naturally infected with C. sinensis. The snails which released more than 1,000 cercariae/day were 30.8% out of total collected. A snail uniquely released 5,840 cercariae/day in this study. 10. The other trematode cercariae besides C. sinensis were also detected, and the rates out of total snails were the cercariae of Losogenes liberum 6.71%, Cyathocotyle orientalis 0.26%, Notocotylus attenuatus 2.52% and Mucobucaris 0.54%, 11. Ten out of 18 species of fresh water fish caught along the river harbored the metacercariae of C, sinensis. The highest rate of metacercarial infection in fish was detected in Pseudorasbora larva (85.9%). The fish mainly eaten by the inhabitants along the Nam-river, and the metacercarial infection rates were: Zacco platypus 8.0%, Hemibarbus sp. 18.2∼26.7%, Gnathepcgcn sp. 37.5%, Ischikauia steenackeri 42.9% and Pseudogobio esocinus 16.7%. 12. Out of 36 P. larva, the number of metacercariae were about 109 (ranEe; 18∼446) per fish and 27 per gram of flesh. The fish caught in spring harbored the highest number of metacercariae. As indicated above, the prevalence of C. sinensis infection in the inhabitants residing around the Nam-river was relatively high. The farther toward the upper stream areas, the higher was the positive rate of C. sinensis infection, and most of clonorchiasis cases were lightly infected. The snail hosts of C. sinensis distributed all around the Nam-river. Several species of freshwater fish were infected with the metacercariae of C. sinensis and the infection rates were relatively high. To prevent C. sinensis infection in the endemic areas, the effective health education system is suggested as a control measure, although mass treatment is also expected to be useful, using chemotherapeutic agents such as "praziquantel", a recently developed anthelminthic for C. sinensis inftraction.

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우리나라 감자에 발생하는 PVY의 병원학적 특성 및 외피단백질 유전자 분석 (Etiological Properties and Coat Protein Gen Analysis of Potato Virus Y Occuring in Potatoes of Korea)

  • 정승룡
    • 한국식물병리학회:학술대회논문집
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    • 한국식물병리학회 1995년도 Proceedings of special lectures on Molecular Biological Approaches to Plant Disease National Agricultural Science and Technology Institute Suwon, Korea
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    • pp.77-96
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    • 1995
  • To obtain basic informations for the improvement of seed potato production in Korea, some etiological properties of potato virus Y(PVY) distributed in the major seed potato production area(Daekwanryeong) were characterized, and the nucleotide and amino acid sequences of the coat protein gene of the PVY strains isolated were analyzed. PVY strains in Daekwonryeong, an alpine area, were identified to be two strains, PVYo and PVYN by symptoms of indicator plants, and their distribution in potato fields was similar. Major symptom on potato varieties by PVY was grouped as either mosaic alone or mosaic accompanied with veinal necrosis in the lower leaves. The symptom occurrence of the two symptoms was similar with Irish Cobbler, but Superior showed a higher rate of mosaic symptom than the other. The PVY strain which was isolated from potato cv. Superior showing typical mosaic symptoms produced symptoms of PVY-O on the indicator plants of Chenopodium amaranticolor, Nicotiana tabacum cv. Xanthi nc and Physalis floridana, but no symptom o Capsicum annum cv. Ace. Moreover, results from the enzyme-linked immunosorbent assay with monoclonal and polyclonal antibodies showed that the isolated PVY reacts strongly with PYV-O antibodies but does not react specifically with PVY-T antibodies. The purified virus particles were flexious with a size of 730$\times$11nm. On the basis of the above characteristics, the strain was identified to be a PVY-O and named as of PVY-K strain. The flight of vector aphids was observed in late May, however, the first occurrence of infected plants was in mid June with the bait plants surrounded with PVY-infected potato plants and early July with the bait plants surrounded with PVY-free potato plants. PVY infection rates by counting symptoms on bait plants (White Burley) were 1.1% with the field surrounded with PVY-free potato plants and 13.7% the fields surrounded with PVY-infected potato plants, showing the effect of infection pressure. The propagated PVY-K strain on tobacco(N. sylvestris) was purified, and the RNA of the virus was extracted by the method of phenol extraction. The size of PVY-K RNA was measured to be 9, 500 nucleotides on agarose gel electrophoresis. The double-stranded cDNAs of PVY-K coat protein(CP) gene derived by the method of polymerase chain reaction were transformed into the competent cells of E. coli JM 109, and 2 clones(pYK6 and pYK17) among 11 clones were confirmed to contain the full-length cDNA. Purified plasmids from pYK17 were cut with Sph I and Xba I were deleted with exonuclease III and were used for sequencing analysis. The PVY-K CP gene was comprised of 801 nucleotides when counted from the clevage site of CAG(Gln)-GCA(Ala) to the stop codon of TGA and encoded 267 amino acids. The molecular weight of the encoded polypeptides was calculated to be 34, 630 daltons. The base composition of the CP gene was 33.3% of adenine, 25.2% of guanine, 20.1% of cytosine and 21.4% of uracil. The polypeptide encoded by PVY-K CP gene was comprised of 22 alanines, 20 threonines, 19 glutamic acids and 18 glycines in order. The homology of nucleotide sequence of PVY-K CP gene with those of PVY-O(Japan), PVY-T(Japan), PVY-TH(Japan), PVYN(the Netherlands), and PVYN(France) was represented as 97.3%, 88.9%, 89.3%, 89.6% and 98.5%, respectively. The amino acid sequence homology of the polypeptide encoded by PVY-K CP gene with those encoded by viruses was represented as 97.4%, 92.5%, 92.9%, 92.9%, and 98.5%, respectively.

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