• 제목/요약/키워드: subtractive

검색결과 289건 처리시간 0.024초

다이아지논에 의해 야기된 송사리의 이상행동 연관 분자생물지표의 선발 (Selection of Molecular Biomarkers Relevant to Abnormal Behaviors of Medaka Fish (Oryzias latipes) Caused by Diazinon)

  • 고성철;신성우;조현덕;전태수;김정상;이성규
    • Environmental Analysis Health and Toxicology
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    • 제24권4호
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    • pp.321-332
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    • 2009
  • 본 연구의 목적은 다이아지논(Diazinon; O, O-diethyl O-[6-methyl-2 (1-methylethyl)-4-pyrimidinyl] phosphorothioate)에 노출된 모델 생물체(송사리)의 행동변화와 관련된 분자생물학적 기전 규명을 통하여 비정상적 행동의 모니터링을 위한 생물지표(biomarker)를 개발하는데 있다. 이를 위해 우선 suppression subtractive hybridization (SSH) 및 DNA microarray 기법을 활용하여 다양한 유전자를 스크리닝하였다. 다이아지논에 노출시킨 송사리에서 발현의 차이가 나는 상향 조절된 유전자 97개 (알려지지 않은 유전자 27개 포함)와 하향 조절된 유전자 99개 (알려지지 않은 유전자 60개 포함)를 동정 하였고 이들 중 이상행동과 관련되는 것으로 보이는 유전자 10개 (상향조절 5개, 하향조절 5개)를 선발하였다. 이들 중에서 primer 제작이 잘된 beta-1, Orla C3-1, parvalbumin 및 apolipoprotein E을 선발하여 그 유전자 발현을 real-time PCR 기법을 사용하여 정량적으로 모니터링 하였다. Orla C3-1, parvalbumin 및 apolipoprotein E는 고농도의 다이아지논 처리(1000 ppb; 24 h)에서 그 발현이 억제됨이 관찰되었다. 다이아지논 처리 시 신경질환 (알츠하이머 병 및 다운신드롬)에 관련된 apolipoprotein E와 근육세포의 유연화에 작용하는 parvalbumin 등의 발현억제는 송사리의 인지능력 교란 및 근육세포의 경직 등을 각각 유도하여 송사리의 비정상적 행동을 야기하는 것으로 판단되었다. 따라서 이들 생물지표는 신경독성물질에 의한 송사리 및 기타 어류의 이상행동의 변화의 감지에 활용될 수 있을 것으로 사료된다.

마이크로 드로플릿 셀 기법과 임계공식온도 측정 기법을 이용한 적층가공 Ti-6Al-4V 합금의 내식성 평가 (Ti-6Al-4V Alloy Fabricated by Additive Manufacturing Method Using Micro-droplet Cell and Critical Pitting Temperature Techniques and Evaluation of its Resistance to Corrosion)

  • 서동일;이재봉
    • Corrosion Science and Technology
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    • 제17권3호
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    • pp.129-137
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    • 2018
  • The resistance to corrosion of additive manufactured (3D printing) Ti-6Al-4V alloys was investigated using micro-electrochemical tests. In terms of corrosion resistance, the acicular martensitic ${\alpha}^{\prime}$ phase in such additive manufactured Ti-6Al-4V was the focus of attention, and its behavior was distinct from that of conventional subtractive manufactured Ti-6Al-4V. To order to identify ${\alpha}^{\prime}$ phase, XRD tests were performed and micro Vickers hardness was measured for different grains (bright and dark grains) in the additive manufactured Ti-6Al-4V alloy. Micro-electrochemical tests were performed to measure corrosion resistance of bright and dark grains in the additive manufactured Ti-6Al-4V alloy with specially designed electrochemical micro-droplet cell. Critical pitting temperature (CPT) measurement was performed to evaluate the resistance to pitting corrosion of additive manufactured Ti-6Al-4V alloys with different volumes of ${\alpha}^{\prime}$ phase and subtractive manufactured Ti-6Al-4V alloy. The dark grains of the laminated Ti-6Al-4V alloy distributed broader than the bright grains measured with low microhardness. The dark grains of the Ti-6Al-4V alloy, which was rich in martensite ${\alpha}^{\prime}$, had lower general corrosion and pitting resistance than bright grains. As the fraction of martensite ${\alpha}^{\prime}$ phase increased, the resistance to the pitting corrosion decreased.

A study on the machining accuracy of dental digital method focusing on dental inlay

  • Bae, Eun-Jeong;Jeong, Il-Do;Kim, Woong-Chul;Kim, Ji-Hwan
    • The Journal of Advanced Prosthodontics
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    • 제10권4호
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    • pp.321-327
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    • 2018
  • PURPOSE. The purpose of this study was to compare the cutting method and the lamination method to investigate whether the CAD data of the proposed inlay shape are machined correctly. MATERIALS AND METHODS. The Mesial-Occlusal shape of the inlay was modeled by changing the stereolithography (STL). Each group used SLS (metal powder) or SLA (photocurable resin) in the additive method, and wax or zirconia in the subtractive method (n=10 per group, total n=40). Three-dimensional (3D) analysis program (Geomagic Control X inspection software; 3D systems) was used for the alignment and analysis. The root mean square (RMS) in the 2D plane state was measured within $50{\mu}m$ radius of eight comparison measuring points (CMP). Differences were analyzed using one-way analysis of variance and post-hoc Tukey's test were used (${\alpha}=.05$). RESULTS. There was a significant difference in RMS only in SLA and SLS of 2D section (P<.05). In CMP mean, CMP 4 ($-5.3{\pm}46.7{\mu}m$) had a value closest to 0, while CMP 6 ($20.1{\pm}42.4{\mu}m$) and CMP 1 ($-89.2{\pm}61.4{\mu}m$) had the greatest positive value and the greatest negative value, respectively. CONCLUSION. Since the errors obtained from the study do not exceed the clinically acceptable values, the lamination method and the cutting method can be used clinically.

Screening of Ecotoxicant Responsive Genes and Expression Analysis of Benzo[a]pyrene-exposed Rockfish (Sebastes schlgeli)

  • Yum, Seung-Shic;Woo, Seon-Ock;Lee, Taek-Kyun
    • Molecular & Cellular Toxicology
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    • 제2권2호
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    • pp.114-119
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    • 2006
  • Benzo[a]pyrene is a representative ecotoxicant in marine environment and a model compound of polycyclic aromatic hydrocarbons, which has an ability to bioaccumulate in aquatic organisms. This study aimed to identify molecular biomarkers suitable for assessing environmental pollution using a microarray technique. We examined the effects of benzo[a]pyrene on gene expressions in the rockfish, Sebastes schlegeli. We constructed the subtractive cDNA library with hepatic RNA from benzo[a]pyrene-exposed and non-exposed control fish. From the library 10,000 candidate clones were selected randomly and cDNA microarray was constructed. We determined benzo[a]pyrene-responsive genes using a high-density microarray. Statistical analysis showed that approximately 400 genes are significantly induced or reduced by benzo[a]pyrene treatment ($2\;{\mu}m$). Especially gene expression changes of 4 candidate clones among the up- or down-regulated genes were investigated in 6, 12 and 24 hr BaP-exposed fish groups. Many methods have been developed to monitor marine environmental status, which depend on quantifying the levels of the toxic components in polluted seawater or on ecological accessing, such as species diversity or richness. However, those methods could not provide information on physiological or genetic changes induced by such environmental stresses. Comparing with the conventional methods, these data will propose that benzo[a]pyrene-responsive genes can be useful for biological risk assessment of polycyclic aromatic hydrocarbons on marine organism at molecular level.

Evaluation of marginal discrepancy of pressable ceramic veneer fabricated using CAD/CAM system: Additive and subtractive manufacturing

  • Kang, Seen-Young;Lee, Ha-Na;Kim, Ji-Hwan;Kim, Woong-Chul
    • The Journal of Advanced Prosthodontics
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    • 제10권5호
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    • pp.347-353
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    • 2018
  • PURPOSE. The purpose of this study was to evaluate the marginal discrepancy of heat-pressed ceramic veneers manufactured using a CAD/CAM system. MATERIALS AND METHODS. The ceramic veneers for the abutment of a maxillary left central incisor were designed using a CAD/CAM software program. Ten veneers using a microstereolithography apparatus (AM group), ten veneers using a five-axis milling machine (SM group), and ten veneers using a traditional free-hand wax technique (TW group) were prepared according to the respective manufacturing method. The ceramic veneers were also fabricated using a heat-press technique, and a silicone replica was used to measure their marginal discrepancy. The marginal discrepancies were measured using a digital microscope (${\times}160$ magnification). The data were analyzed using a nonparametric Kruskal-Wallis H test. Finally, post-hoc comparisons were conducted using Bonferroni-corrected Mann-Whitney U tests (${\alpha}=.05$). RESULTS. The $mean{\pm}SD$ of the total marginal discrepancy was $99.68{\pm}28.01{\mu}m$ for the AM group, $76.60{\pm}28.76{\mu}m$ for the SM group, and $83.08{\pm}39.74{\mu}m$ for the TW group. There were significant differences in the total marginal discrepancies of the ceramic veneers (P<.05). CONCLUSION. The SM group showed a better fit than the AM and TW groups. However, all values were within the clinical tolerance. Therefore, CAD/CAM manufacturing methods can replace the traditional free-hand wax technique.

MC3T3-E1 osteoblast adhesion to laser induced hydroxyapatite coating on Ti alloy

  • Huang, Lu;Goddard, Samuel C.;Soundarapandian, Santhanakrishnan;Cao, Yu;Dahotre, Narendra B.;He, Wei
    • Biomaterials and Biomechanics in Bioengineering
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    • 제1권2호
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    • pp.81-93
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    • 2014
  • An in vitro cell study evaluating cell adhesion to hydroxyapatite (HA) coated prosthetic Ti-6Al-4V alloy via laser treatment is presented in comparison with uncoated alloy. Based on our previous in vitro biocompatibility study, which demonstrated higher cell attachment and proliferation with MC3T3-E1 preosteoblast cells, the present investigation aims to reveal the effect of laser coating Ti alloy with HA on the adhesion strength of bone-forming cells against centrifugal forces. Remaining cells on different substrates after centrifugation were visualized using fluorescent staining. Semi-quantifications on the numbers of cells were conducted based on fluorescent images, which demonstrated higher numbers of cells retained on HA laser treated substrates post centrifugation. The results indicate potential increase in the normalized maximum force required to displace cells from HA coated surfaces versus uncoated control surface. The possible mechanisms that govern the enhancing effect were discussed, including surface roughness, chemistry, wettability, and protein adsorption. The improvement in cell adhesion through laser treatment with a biomimetic coating could be useful in reducing tissue damage at the prosthetic to bone junction and minimizing the loosening of prosthetics over time.

Identification of p54nrb and the 14-3-3 Protein HS1 as TNF-α-Inducible Genes Related to Cell Cycle Control and Apoptosis in Human Arterial Endothelial Cells

  • Stier, Sebastian;Totzke, Gudrun;Grunewald, Elisabeth;Neuhaus, Thomas;Fronhoffs, Stefan;Schoneborn, Silke;Vetter, Hans;Ko, Yon
    • BMB Reports
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    • 제38권4호
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    • pp.447-456
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    • 2005
  • TNF-$\alpha$ plays a pivotal role in inflammation processes which are mainly regulated by endothelial cells. While TNF-$\alpha$ induces apoptosis of several cell types like tumor cells, endothelial cells are resistant to TNFa mediated cell death. The cytotoxic effects of TNF-$\alpha$ on most cells are only evident if RNA or protein synthesis is inhibited, suggesting that de novo RNA or protein synthesis protect cells from TNF-$\alpha$ cytotoxicity, presumably by NF-${\kappa}B$ mediated induction of protective genes. However, the cytoprotective genes involved in NF-${\kappa}B$ dependent endothelial cell survival have not been sufficiently identified. In the present study, the suppression subtractive hybridization (SSH) method was employed to identify rarely transcribed TNF-$\alpha$ inducible genes in human arterial endothelial cells related to cell survival and cell cycle. The TNF-$\alpha$-induced expression of the RNA binding protein $p54^{nrb}$ and the 14-3-3 protein HS1 as shown here for the first time may contribute to the TNF-$\alpha$ mediated cell protection of endothelial cells. These genes have been shown to play pivotal roles in cell survival and cell cycle control in different experimental settings. The concerted expression of these genes together with other genes related to cell protection and cell cycle like DnaJ, $p21^{cip1}$ and the ubiquitin activating enzyme E1 demonstrates the identification of new genes in the context of TNF-$\alpha$ induced gene expression patterns mediating the prosurvival effect of TNF-$\alpha$ in endothelial cells.

Comparative Analysis of Gene Expression in the Female Reproductive Organs

  • Kim, Min-Goo;Seo, Hee-Won;Choi, Yo-Han;Lee, Chang-Kyu;Ka, Hak-Hyun
    • 한국수정란이식학회지
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    • 제24권2호
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    • pp.77-87
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    • 2009
  • To understand molecular and cellular mechanisms of many gene products in the female reproductive organs including the ovary and uterine endometrium as well as during embryo development, researchers have developed and utilized many effective methodologies to analyze gene expression in cells, tissues and animals over the last several decades. For example, blotting techniques have helped to understand molecular functions at DNA, RNA and protein levels, and the reverse transcription-polymerase chain reaction (RT-PCR) method has been widely used in gene expression analysis. However, some conventional methods are not sufficient to understand regulation and function of genes expressed in very complex patterns in many organs. Thus, it is required to adopt more high-throughput and reliable techniques. Here, we describe several techniques used widely recently to analyze gene expression, including annealing control based-PCR, differential display-PCR, expressed sequence tag, suppression subtractive hybridization and microarray techniques. Use of these techniques will help to analyze expression pattern of many genes from small scale to large scale and to compare expression patterns of genes in one sample to another. In this review, we described principles of these methodologies and summarized examples of comparative analysis of gene expression in female reproductive organs with help of those methodologies.

열 가압 방식을 사용하여 제작된 인레이 세라믹 수복물의 적합도 평가 (Assessment of inlay ceramic restorations manufactured using the hot-pressing method)

  • 이범일;유승규;유승민;박동인;김지환
    • 대한치과기공학회지
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    • 제42권1호
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    • pp.9-16
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    • 2020
  • Purpose: The purpose of this study was to compare the marginal and internal fit of lithium disilicate ceramic inlay produced by heat pressing that inlay pattern made by subtractive manufacturing and additive manufacturing method. Methods: A mandibular lower first molar that mesial occlusal cavity (MO cavity) die was prepared. After fabricating an epoxy resin model using a silicone impression material, epoxy resin die was scanned with a dental model scanner to design an MO cavity inlay. The designed STL pile was used to fabricate wax patterns and resin patterns, and then lithium disilicate ceramic inlays were fabricated using hot-press method. For the measurement of the marginal and internal gap of the lithium disilicate, silicone replica method was applied, and gap was measured through an optical microscope (x 80). Data were tested for significant differences using the Mann-Whitney Utest. Results: The marginal fit was 103.56±9.92㎛ in the MIL-IN group and 81.57±9.33㎛ in the SLA-IN group, with a significant difference found between the two groups (p<0.05). The internal fit was 120.99±17.52㎛ in the MIL-IN group and 99.18±6.65㎛ in the SLA-IN group, with a significant difference found between the two groups (p<0.05). Conclusion: It is clinically more appropriate to apply the additive manufacturing than subtractive manufacturing method in producing lithium disilicate inlay using CAD/CAM system.

The Determination of Elementary School Students' Successes in Choosing an Operation and the Strategies They Used While Solving Real-World Problems

  • Soylu, Yasin
    • 한국수학교육학회지시리즈D:수학교육연구
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    • 제11권4호
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    • pp.247-263
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    • 2007
  • Problem solving takes place not only in mathematics classes but also in real-world. For this reason, a problem and the structure of problem solving, and the enhancing of success in problem solving is a subject which has been studied by any educators. In this direction, the aim of this study is that the strategy used by students in Turkey when solving oral problems and their achievements of choosing operations when solving oral problems has been researched. In the research, the students have been asked three types of questions made up groups of 5. In the first category, S-problems (standard problems not requiring to determine any strategy but can be easily solved with only the applications of arithmetical operations), in the second category, AS-SA problems (problems that can be solved with the key word of additive operation despite to its being a subtractive operation, and containing the key word of subtractive operation despite to its being an additive operation), and in the third category P-problems (problematic problem) take place. It is seen that students did not have so much difficulty in S-problems, mistakes were made in determining operations for problem solving because of memorizing certain essential concepts, and the succession rate of students is very low in P-problems. The reasons of these mistakes as a summary are given below: $\cdot$ Because of memorizing some certain key concepts about operations mistakes have been done in choosing operations. $\cdot$ Not giving place to problems which has no solution and with incomplete information in mathematics. $\cdot$ Thinking of students that every problem has a solution since they don't encounter every type of problems in mathematics classes and course books.

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