• Title/Summary/Keyword: Molecular techniques

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The Detection and Diagnosis Methods of Infectious Viroids caused Plant Diseases (식물체에 감염성 질병을 유발하는 바이로이드 검출 및 진단 방법)

  • Lee, Se Hee;Kim, Yang-Hoon;Ahn, Ji-Young
    • Journal of Life Science
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    • v.26 no.5
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    • pp.620-631
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    • 2016
  • Viroids are about 250-400 base pair of short single strand RNA fragments have been associated with economically important plant diseases. Due to the lack of protein expression capacity associated with replication, it is very difficult to diagnosis viroid diseases in serological methods. For detecting viroid at plants, molecular-based techniques such as agarose gel electrophoresis, polyacrylamide gel electrophoresis (PAGE), DNA-hybridization, blotting analysis and conventional RT-PCR are reliable. Real-time RT-PCR methods that grafted on RT-PCR methods with improved confirmation methods have been also utilized. However, they are still labor-intensive, time-consuming, and require personnel with expertise. Loop-mediated Isothermal Amplification (LAMP) method is a nucleic acid amplification method under the isothermal condition. The LAMP methodology has been reported to be simple, rapid, sensitive and field applicable in detecting a variety of pathogens. The results of LAMP method can be colorized by adding a visible material such as SYBR green I, Evagreen, Calcein, Berberine and Hydroxy naphthol blue (HNB) with simple equipment or naked eyes. The combination of LAMP method and nucleic pathogens, viroids, can be used to realize simple diagnosis platform for the genetic point-of care testing system. The aim at this review is to summary viroid-caused diseases and the simple visible approach for diagnosing viroids using Loop-mediated Isothermal Amplification (LAMP) method.

Purification and Characterization of Polyphenol Oxidase from Oyster Mushroom (Pleurotus ostreatus) (느타리버섯(Pleurotus ostreatus)의 Polyphenol Oxidase 분리 정제 및 특성 조사)

  • Choi, Ju-Hee;Kim, Hyun-Jin;Park, Sun-Young;Ham, Kyung-Sik
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.40 no.10
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    • pp.1447-1452
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    • 2011
  • Polyphenol oxidase (PPO) isoforms were partially purified from oyster mushroom (Pleurotus ostreatus) using various chromatography techniques, and their characteristics of heat stability, substrate affinity, optimum pH, and optimum temperature were investigated. Three PPO isoforms named PO-I, PO-II-1, and PO-II-2 were partially purified from oyster mushroom. The molecular weight of PO-II-1 was 70 kDa and PO-I and PO-II-2 were less than 6 kDa each. Characterization was carried out using a PPO isoform partially purified by hydrophobic interaction chromatography. Optimum temperature was $55^{\circ}C$ and optimum pH 5.0. However, the PPO was inactivated at neutral pH or by heating at $80^{\circ}C$ for 30 min, while the 40% PPO still remained active after heating at $60^{\circ}C$ for 45 min. The PPO isoform showed the highest substrate affinity to chlorogenic acid and pyrogallol, in which KM values were 1.01 and 2.06 mM, respectively. Therefore, these results suggested that the mushrooms should be stored at a pH higher than 7.0 and at a low temperature to prevent enzymatic browning.

Review and application of environmental DNA (eDNA) investigation of terrestrial species in urban ecosystem (도시 내 육상 생물종 모니터링을 위한 환경DNA 리뷰 및 적용)

  • Kim, Whee-Moon;Kim, Seoung-Yeal;Park, Il-Su;Lee, Hyun-Jung;Kim, Kyeong-Tae;Kim, Young;Kim, Hye-Joung;Kwak, Min-Ho;Lim, Tae-Yang;Park, Chan;Song, Won-Kyong
    • Journal of the Korean Society of Environmental Restoration Technology
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    • v.23 no.2
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    • pp.69-89
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    • 2020
  • Scientific trust and quantification of traditional species investigation and results that have been used in ecology for decades has always been a problem and concern for ecologists. Global ecologists have proposed DNA-based species investigation studies to find answers to problems. In this study, we reviewed the global trend of research on environmental DNA(eDNA), which is a method for monitoring species by detecting DNA of organisms naturally mixed in environmental samples such as water, soil, and feces. The first eDNA research confirmed the possibility of species investigation at the molecular level, and commercialization of NGS(Next Generation Sequencing) and DNA metabarcoding elicits efficient and quantitative species investigation results, and eDNA research is increasing in the filed of ecology. In this study, mammals and birds were detected using MiMammal universal primers from 23 samples(3 natural reserves; 20 water bowls) out of 4 patches to verify eDNA for urban ecosystems in Suwon, and eDNA was verified by performing camera trapping and field survey. Most terrestrial species were detected through eDNA, and particularly, mice(Mus musculus), and Vinous-throated Parrotbill (Sinosuthora webbiana) were identified only with eDNA, It has been confirmed to be highly effective by investigating techniques for small and internal species. However, due to the lack of resolution of the primer, weasels(Mustela sibirica) and squirrels(Melanochromis auratus) were not detected, and it was confirmed that the traditional investigation method was effective only for a few species, such as Mogera robusta(Mogera robusta). Therefore, it is judged that the effects of species investigation can be maximized only when eDNA is combined with traditional field survey and Camera trapping to complement each other.

Maximization of The Number of Follicular Oocytes Recovered from The Bovine Ovaries (소 난소로부터 회수난포란수의 극대화 방법)

  • 유형진;최승철;이상호
    • Korean Journal of Animal Reproduction
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    • v.17 no.2
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    • pp.149-157
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    • 1993
  • A new technique was established to maximize the numbers of follicular oocytes recovered from the ovaries obtained at the slaughter house. And their further developmental capacity was demonstrated. There recovery techniques were used; aspiration (ASP, control), slicing (SLC) and slicing combining aspiration (ASP+SLC). Recovered oocytes were cultured in TCM 199+15% FCS+gonadotrophins in an atmosphere of 5% CO$_2$ in air at 39$^{\circ}C$ for 24 h. The nuclear maturation was detemined with chromo-some configuration by rapid staining. And cytoplasmic maturation was examined by the formation of female pronuclei with parthenogenetic activation of the matured oocyte after 18 h of co-culture with granulosa cell monolayer. Total 1,641 bovine follicular oocytes recovered from 245 ovaries. The number of oocytcs per ovary was 1.87 in ASP, 11.05 in SLC and 7.88 in ASP+SLC, respectively. SLC would yield 5.9 folds increase, compared with ASP. The rate of maturation were 92.9% in ASP, 79.1% in SLC and 71.7% in ASP+SLC, respectively. Although the maturation rate in ASP was the highest, metaphase II oocytes per ovary in SLC was 5 times higher than that of ASP. The rates of pronuclei formation upon ethanol activation were 75% in ASP, 67% in SLC and 62.5% in ASP+SLC, respectively. The results demonstrate that it should be possible to maximize the number of the follicular oocyte from the ovary for mass production of bovine embryos. Thus the established technique may provide efficient supply of bovine embryos for biochemical and molecular study of early bovine embryos.

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Characterization of Poly(methyl methacrylate)-tin (IV) Chloride Blend by TG-DTG-DTA, IR and Pyrolysis-GC-MS Techniques

  • Arshad, Muhammad;Masud, Khalid;Arif, Muhammad;Rehman, Saeed-Ur;Saeed, Aamer;Zaidi, Jamshed Hussain
    • Bulletin of the Korean Chemical Society
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    • v.32 no.9
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    • pp.3295-3305
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    • 2011
  • Thermal behavior of poly (methyl methacrylate) was analyzed in the presence of tin (IV) chloride. Five different proportions - polymer to additive - were selected for casting films from common solvent. TG, DTG and DTA were employed to monitor thermal degradation of the systems. IR and py-GC-MS helped identify the decomposition products. The blends start degrading at a temperature lower than that of the neat polymer and higher than that of the pure additive. Complex formation between tin of additive and carbonyl oxygen (pendent groups of MMA units) was noticed in the films soon after the mixing of the components in the blends. The samples were also heated at three different temperatures to determine the composition of residues left after the expulsion of volatiles. The polymer, blends and additive exhibited a one step, two-step and three-step degradation, respectively. $T_0$ is highest for the polymer, lowest for the additive and is either $60^{\circ}C$ or $70^{\circ}C$ for the blends. The amount of residue increases down the series [moving from blend-1 (minimum additive concentration) to blend-5 (maximum additive concentration)]. For blend-1, it is 7% of the original mass whereas it is 16% for blend-5. $T_{max}$ also goes up as the concentration of additive in the blends is elevated. The complexation appears to be the cause of observed stabilization. Some new products of degradation were noted apart from those reported earlier. These included methanol, isobutyric acid, acid chloride, etc. Molecular-level mixing of the constituents and "positioning effect" of the additive may have brought about the formation of new compounds. Routes are proposed for the appearance of these substances. Horizontal burning tests were also conducted on polymer and blends and the results are discussed. Activation energies and reaction orders were calculated. Activation energy is highest for the polymer, i.e., 138.9 Kcal/mol while the range for blends is from 51 to 39 Kcal/mol. Stability zones are highlighted for the blends. The interaction between the blended parts seems to be chemical in nature.

Wireless Energy and Data Transmission Using Inductive Coupling (유도결합방식에 의한 무선 에너지 및 데이터 전송)

  • Lee, Joon-Ha
    • Progress in Medical Physics
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    • v.19 no.1
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    • pp.42-48
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    • 2008
  • Bio-implantable devices such as heart pacers, gastric pacers and drug-delivery systems require power for carrying out their intended functions. These devices are usually powered through a battery implanted with the system or are wired to an external power source. This paper describes an inductive power transmission link, which was developed for an implantable stimulator for direct stimulation of denervated muscles. The carrier frequency is around 1MHz, the transmitter coil has a diameter of 46mm, and the implant coil is 46mm. Data transmission to the implant with amplitude shift keying (ASK) and back to the transmitter with passive telemetry can be added without major design changes. We chose the range of coil spacing (2 to 30mm) to care for lateral misalignment, as it occurs in practical use. If the transmitter coil has a well defined and reliable position in respect to the implant, a smaller working range might be sufficient. Under these conditions the link can be operated in fixed frequency mode, and reaches even higher efficiencies of up to 37%. The link transmits a current of 50 mA over a distance range of 2-15 mm with an efficiency of more than 20% in tracking frequency. The efficiency of the link was optimized with different approaches. A class E transmitter was used to minimize losses of the power stage. The geometry and material of the transmitter coil was optimized for maximum coupling. Phase lock techniques were used to achieve frequency tracking, keeping the transmitter optimally tuned at different coupling conditions caused by coil distance variations.

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A Study on Development of Experimental Contents Using 3-channel Multi-Image Playback Technique: Based on transparent OLED and dual layer display system (3채널 멀티 영상 재생 기법과 증강현실을 이용한 체험 콘텐츠 제작에 관한 연구: 투명 OLED 및 듀얼 레이어 디스플레이 시스템 기반)

  • Lee, Sang-Hyun
    • The Journal of the Institute of Internet, Broadcasting and Communication
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    • v.17 no.6
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    • pp.151-160
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    • 2017
  • Among the methods of developing tourist spots and culture as the experience contents, it is a common method to display high-quality video images on a large display, and it is necessary to make a special difference between the participant's active participation and the visual experience in other regions. In this paper, using the single molecular OLED and active type, the regional tourist spots blend transparent OLED dual-layer display systems with the extended image implementation and augmented interaction techniques to give the participants a real-world experience, such as directing to new experiences and beautiful sights. In this paper, additional images and UI layers are applied to the layers of the images to allow visitors to experience sightseeing information, weather, maps, accommodations, festivals and photo materials with image. In addition to the dual-layer system, it also added a multi-display system that additionally has one vertical 55-inch display on each side, adding to the experience the immersive experience and interface interlocking fun. By using transparent OLED, dual layer panel and 3-channel Multi-image playback technique, the augmented type experience contents which can experience the local attractions in Jeollanamdo province in Korea at all time without any limitation of time and space were developed.

Performance analysis of volleyball games using the social network and text mining techniques (사회네트워크분석과 텍스트마이닝을 이용한 배구 경기력 분석)

  • Kang, Byounguk;Huh, Mankyu;Choi, Seungbae
    • Journal of the Korean Data and Information Science Society
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    • v.26 no.3
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    • pp.619-630
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    • 2015
  • The purpose of this study is to provide basic information to develop a game strategy plan of a team in a future by identifying the patterns of attack and pass of national men's professional volleyball teams and extracting core key words related with volleyball game performance to evaluate game performance using 'social network analysis' and 'text mining'. As for the analysis result of 'social network analysis' with the whole data, group '0' (6 players) and group '1' (11 players) were partitioned. A point of view the degree centrality and betweenness centrality in 'social network analysis' results, we can know that the group '1' more active game performance than the group '0'. The significant result for two group (win and loss) obtained by 'text mining' according to two groups ('0' and '1') obtained by 'social network analysis' showed significant difference (p-value: 0.001). As for clustering of each network, group '0' had the tendency to score points through set player D and E. In group '1', the player K had the tendency to fail if he attack through 'dig'; players C and D have a good performance through 'set' play.

A study of Expression of TGF-β1, c-Myc, Erb-B2 and Thymosin-β4 Gene in Alcoholic Liver Damage Tissue. (알코올성 간 손상 조직에서 TGF-β1와 c-Myc, Erb-B2, Thymosin-β4 유전자 발현 융합 연구)

  • Kim, Jean-Soo;Choi, Sang-Ki
    • Journal of the Korea Convergence Society
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    • v.9 no.5
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    • pp.91-97
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    • 2018
  • This study has been conducted to see the expression of $TGF-{\beta}_1$, c-Myc, Erb-B2 and $Thymosin-{\beta}_4$ genes in ethanol - damaged liver tissues. Experimental groups were divided into 2 groups, one where damaged liver was caused by 25% ethanol and normal group administered with purified water. Results of test showed the expression of $TGF-{\beta}_1$, c-Myc, and $Thymosin-{\beta}_4$ genes was higher in the experimental group treated with 25% ethanol than in the normal group. Erb-B2 gene was not expressed clearly. Thus, it is considered that we can expect to utilize $TGF-{\beta}_1$, c-Myc 및 $Thymosin-{\beta}_4$ as auxiliary data and find clinical meanings of diagnosis on hepatic diseases, In addition to serologic and histological examination by convergence examining the gene expression status by molecular diagnostic techniques in liver-related disease prevention and diagnosis through results of this study.

The Endotoxin Assay of Contaminated Titanium Implants following Various Techniques of Detoxification (오염된 임프란트 표면의 해독 방법에 따른 내독소 제거 효과에 관한 연구)

  • Park, Joong-Hee;Lim, Sung-Bin
    • Journal of Periodontal and Implant Science
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    • v.34 no.1
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    • pp.71-81
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    • 2004
  • Peri-implantitis could be the result of biomechanical and occlusal overload as well as microbiologic invasion. The dental implant may be more susceptible to dental plaque than the natural tooth, as the predictability of a stable soft tissue attachment complex has not yet been confirmed. With the development of peri-implantitis, the implant surface would be exposed to the oral environment and becomes coated with bacteria. The objective of therapy for this condition is to regain integration of the implant with bone. Since fibroblast adherence to surfaces is impeded by endotoxin, it would seem that decontamination would be desirable to obtain maximum osseointegration. The purpose of this study was to determine whether various chemotherapeutic and mechanical treatments(distilled water, air-powder abrasive, hypersaturated citric acid, tetracycline) can detoxify contaminated titanium implant surface by means of kinetic LAL test. Experimental rough surface titanium disks were fabricated. All of them were divided into two groups(A.a group and P.g group) and each contaminated by A. actinomycetemcomitans and P. gingivalis suspension. Contaminated disks were treated with distilled water, air-powder abrasive, citric acid and tetracycline, and then all disks were placed into LPS-free water for elution. The results were as follows : 1. In A.a group, LPS elute level of all test groups were significantly lower than control group(p<0.05). 2. In A.a group, LPS elute level of test 2, test 3 and test4 groups were significantly lower than that of control group(p<0.05). But, among the test 2, test 3, test4 groups, the significant differences were not detected. 3. In P.g group, LPS elute level of test 2, test 3 and test 4 groups were lower than that of control group(p<0.05). But, among the test groups, the significant differences were not detected. From the result of this study, it would be concluded that air-powder abrasive, hypersaturated citric acid and tetracycline treatments may be effective at reducing endotoxin level on rough titanium implant surfaces, and can be clinically used. But the treatments in peri-implantitis differentially impact osseointegration making one method clinically superior. To gain this knowledges, further molecular biologic and histopathologic studies should be developed.