• Title/Summary/Keyword: double-counting

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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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    • v.14 no.4
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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.

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

  • Kim, Kyeong Nam;Lee, Sun Jeoung;Kim, Raehyun;Son, Yeong Mo
    • Korean Journal of Agricultural and Forest Meteorology
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    • v.16 no.4
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    • pp.336-342
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    • 2014
  • This study was conducted to estimate of the greenhouse gas inventory on forest land at provincial level. The greenhouse gas emissions are calculated according to the K-MRV guidance. We collected activity data from statistical yearbook of forestry and used default emission factors. The annual total $CO_2$ emission in forest land was -58,711Gg $CO_2eq.$ and the annual $CO_2$ emission in loss such as fellings, fuelwood and fire was 19,896Gg $CO_2eq.$ in 2011. The results showed the removals of carbon dioxide in the forest land, it's amount was -38,815Gg $CO_2eq.$ in 2011. Annual net $CO_2$ removal of local forest was highest in Kangwon province in 2011. Our study did not use the many statistics due to exclusion of double counting. There are need complementary activity data and emission factors, and then we will find a way to calculate the greenhouse gas emissions/removals in the near future.

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

  • Jung, Chan-Mi;Shin, Dong-hee
    • Journal of the Korean Society of Earth Science Education
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    • v.10 no.3
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    • pp.235-253
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    • 2017
  • The purpose of this study is to compare by learner groups(K-12, geology-related majoring students, science teachers) how geologic fieldwork education cases in domestic and foreign papers for recent 20 years reflect the elements of inquiry in field geology. The total number of analyzed cases is 53(58 for double counting), and the analysis was conducted on the elements of inquiry in field geology and their sub-element. As a result, there was a clear difference between the cases of geologic fieldwork education for K-12 and college students majoring in geology-related disciplines, in the way of reflecting elements of inquiry in field geology. Because most of K-12 target fieldworks were designed based on the curriculum, it mainly included 2-3 elements of observations, representations, and/or abductive reasoning. On the other hand, because fieldworks for geology-related major students aim to train geologic experts, it diversely contained four or five of the elements of inquiry in field geology, including spatial thinking and diachronic thinking in a high proportion, and also frequently used activities that require specialized skills such as geological mapping. Fieldworks for science teachers were found to have mixed features of K-12 and geology-related major students. Considering the diversity of included inquiry elements, similarities with the activities performed by geologists, and the autonomy of learners, it was analyzed that geologic fieldwork for geology-related major students was more close to authentic geologic inquiry than fieldwork for others. Based on the results of this study, we suggested implications for improving geological fieldwork as authentic science inquiry.

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
    • Proceedings of the Korean Society of Medical Physics Conference
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    • 2002.09a
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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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Detection and quantitation of Bacillus cereus, Staphylococcus aureus, Salmonella Typhimurium and Escherichia coli O157:H7 by droplet digital PCR (Droplet Digital PCR을 이용한 Bacillus cereus, Staphylococcus aureus, Salmonella Typhimurium과 Escherichia coli O157:H7의 검출 및 정량)

  • Kim, Jin-Hee;Yoon, JinSun;Lee, Da-Young;Kim, Dongho;Oh, Se-Wook
    • Korean Journal of Food Science and Technology
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    • v.48 no.5
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    • pp.454-460
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    • 2016
  • In this study, we investigated the possibility of Droplet digital PCR (ddPCR) for detection of foodborne pathogens. ddPCR combines partitioning of PCR reactions into several thousands or millions of individual droplets in a water-oil emulsion, and counting of positive PCR reaction using flow cytometry. Four species of foodborne pathogens, Bacillus cereus, Staphylococcus aureus, Salmonella Typhimurium and Escherichia coli O157:H7, were used to quantify the target sequence with each of the designed primers and double stranded DNA-binding Evagreen dye. All tested foodborne pathogens showed a detection limit ranging from $100fg/{\mu}L$ to $10ng/{\mu}L$. It was concluded that ddPCR could be used to detect very low concentrations of foodborne pathogens from complex food matrices. For multi-detection of target pathogens, we also tested the samples using multiplex ddPCR and obtained successful results.

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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    • v.34 no.5
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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.

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

  • 배경훈;안영겸
    • Parasites, Hosts and Diseases
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    • v.21 no.2
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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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Etiological Properties and Coat Protein Gen Analysis of Potato Virus Y Occuring in Potatoes of Korea (우리나라 감자에 발생하는 PVY의 병원학적 특성 및 외피단백질 유전자 분석)

  • ;Richard M. Bostock
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 1995.06b
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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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