• Title/Summary/Keyword: Direct extraction

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Population-Based Newborn Hearing Impairment Screening Test Using GJB2 Mutation Analysis

  • Lee, Kyung-Ok;Jeong, Su-Jin;Byun, Ji-Young;Kim, Jeong-Sook;Lee, Hye-Jung;Seong, Hye-Soon;Kim, Kyung-Tae
    • Korean Journal of Clinical Laboratory Science
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    • v.39 no.2
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    • pp.113-121
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    • 2007
  • Hearing loss is a common congenital disorder that is frequently associated with mutations in the Cx26 gene (GJB2). Recently, the mutation analysis of GJB2 has been used in a newborn screening test for the detection of hearing impairment. Population-based studies should be performed before the application of genetic testing for the identification of deaf newborns. In this study, 8 positions of GJB2 mutations-including 35delG, 167delT, 235delC, V27I, V37I, M34T, E114G, and I203T-were analyzed using PCR-direct sequencing in a total of 437 healthy Korean neonates. DNAs from dried blood spots were extracted using a commercial DNA extraction kit. The PCR-amplified products (783 bps) of the GJB2 gene were detected using 2% agarose gel electrophoresis and subjected to direct sequencing. The sequences were compared with those in the GenBank database by using the BLAST program. In this study, 5 GJB2 mutations -including V27I (79G>A), V37I (109G>A), E114G (341A>G), I203T (608T>C), and 235delC- were found. Of the 437 neonate samples, 301 subjects showed GJB2 mutations (68.9%, 301/437). The V27I mutation was found in 271 subjects and was the most frequent (62.0%, 271/437). The E114G, I203T and V37I mutations were shown in 146, 17 and 14 subjects, respectively. The 235delC mutation was found in 1 subject. The E114G mutation was frequently accompanied by the V27I mutation. V27I/E114G (97.2%, 143/147) was the most common double mutation and 3 subjects had the double mutation V27I/I203T. A triple mutation, V27I/E114G/I203T, was found in 1 subject. In conclusion, PCR-direct sequencing is a convenient tool for the rapid detection of GJB2 mutations and this data might provide information for the genetic counseling of the GJB2 gene.

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Effects of Flavonoid from Rhus verniciflua on Testosterone Secretion by Rat Leydig Cells In Vitro (옻나무 유래 Flavonoid 처리가 흰쥐 Leydig 세포의 체외배양에서 Testosterone 분비에 미치는 영향)

  • 성환후;최선호;장유민;민관식;우제현;장원경;정남철;나천수;정일정
    • Korean Journal of Animal Reproduction
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    • v.25 no.2
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    • pp.125-130
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    • 2001
  • This study was performed to report a direct dose dependent stimulatory effect of the Flavonoid(F) on basal testosterone secretion and a dose dependent effect on LH induced testosterone production by Leydig cell of matured rats in vitro culture. F was obtained kom the Rhus vernicifua through aceton extraction and silica gel adsorption column chromatography. Leydig cells (1$\times$10$^{6}$ cells/well) from 12 weeks old rats were incubated with or without F(0, 20, 40, 80, 160 ng) or insulin-like growth factor-I(IGF-I) in the presence or absence of LH(10, 100ng). 1. The maximal stimulatory concentrations of testosterone in culture media were showed at 24hr of culture. but these testosterone level were decreased at 36 hr of culture. 2. Flavonoid(80ng) were significantly(P < 0.05) increased testosterone production compared with control groups for 12 hr culture. 3. Testosterone secretion by Leydig cells stimulated with LH(10, 100ng) for 6 hr and 12hr culture compared with 3 hr culture. 4. LH 10 ng augmented testosterone were increased by addition of F 40 ng for 12 hr culture. 5. F(0 and 40 ng) also enhanced LH 10 ng stimulated testosterone for 3 hr Leydig cells culture. 6. Addition of IGF-I 100 ng to the culture medium for 6 hr were increased the concentration of testosterone by Leydig cells stimulated with 100 ng LH. These results indicate that Flavonoid has a direct stimulatory effect on basal testosterone secretion in rat Leydig cells, and also modulates LH mediated testosterone. Therefore, Flavonoid may act as a modulator on gonadal development or gonadal steroidogenesis in direct or indirect.

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Diagnosis of Diabetes Using Voltage Analysis Based on EIS (Electro Interstitial Scan) (EIS 기반 전압신호 분석을 통한 당뇨병 진단 가능성 평가)

  • Bae, Jang-Han;Kim, Soochan;Kaewkannate, Kanitthika;Jun, Min-Ho;Kim, Jaeuk U.
    • Journal of the Institute of Electronics and Information Engineers
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    • v.53 no.11
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    • pp.114-122
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    • 2016
  • EIS (Electro interstitial scan) is a non-invasive and simple method to find the physio-pathological information inferred by electric current response with respect to low direct current applied between remote sites of the body. Although a few EIS-based devices for diagnosing diabetes were commercialized, they were not successful in offering clinical validity nor in confirming diagnostic principle. In this study, we measured the voltage responses of diabetic patients and normal subjects with a commercialized EIS device to test the usefulness of EIS in screening diabetes. For this purpose, voltage was measured between pairs of electrodes contacted at both palm, both soles of the feet and left and right forehead above both eyes. After feature extraction of voltage signals, the AUC (area under the curve) between the two groups was calculated and we found that seven variables were appropriately shown above 60% of accuracy. In addition, we applied the k-NN (k-nearest neighbors) method and found that the accuracy of classification between the two groups reached the accuracy of 76.2%. This result implies that the voltage response analysis based on EIS has potential as a diabetics screening method.

Production and Action of Microbial Piscicidal Substance (미생물에 의한 살어성물질의 생성 및 그 작용)

  • 도재호;서정훈
    • Microbiology and Biotechnology Letters
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    • v.6 no.1
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    • pp.41-46
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    • 1978
  • Piscicidal substance produced by Streptomyces sp. isolated from soil was toxic against various kinds of fish. After extraction with CH$Cl_3$ from the culture medium, the substance was purified by avicel column chromatography. In order to test toxicity, various kinds of fish were subjected to the acqueous solution of 100 us of the substance per liter of water. Generally, the substance was toxic to most fish, but Macropodus chinenes and Misgurnus mizolepis are resistant to the substance than Gobius similis and Pseudorasbora parva. The substance was stable at pH range, 3.0 to 7.0, but labile at alkaline pH, and to heat as well. Succinic dehydrogenase on most of tissue cell of Cyprinus carpio was inhibited by this substance strongly, but spinal cord was not inhibited. By addition of Cu and Pb salts to the culture medium, piscicidal substance producibility was activated.

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Mass-Production of Acetylcholinesterase Sensitive to Organophosphosphates and Carbamates Insecticides (유기인계 및 카바메이트계 농약의 고감수성 아세틸콜린에스테라이즈의 대량생산)

  • Kim, Young-Mee;K., Cho;Cho, Moon-Jae
    • Applied Biological Chemistry
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    • v.46 no.4
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    • pp.353-360
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    • 2003
  • For the simple rapid bioassay of organophosphorus and carbamate pesticide residues, a mass-production system of acetycholinesterase (AChE, EC 3.1.1.7, MAChE) using baculovirus and insect cell culture was constructed. The cDNA for AChE was synthesized from Drosophila melanogaster in Halla Mountain, the lipid anchor tail was removed by PCR and was used for the site-directed mutagenesis of three amino acid residues (E107Y, F368L, L408F). The mutated cDNA was inserted into the baculovirus vector and expressed in insect cells. Maximum cell growth and enzyme activity were reached when the cells $(2{\times}10^6\;cell/ml)$ were infected four times at four-day-intervals. His-tag containing MAChE was purified using Ni-NTA column and used for characterization. The activity was maintained under various pHs (3-10) and temperatures $(20-50^{\circ}C)$ under experimental conditions. As an extraction solution for pesticides, methanol is more effective than ethanol. Against major organophosphate and carbamate pesticides, the MAChE showed better sensitivity than AChE and AChE from housefly (Taiwan).

Matrix effect on the Determination of Inorganic Priority Pollutants in Sludges (오니 시료중의 무기 Priority Pollutants의 분석 과정에 미치는 매질의 영향)

  • Lee, Huk-Hee;Lee, Sueg-Geun
    • Analytical Science and Technology
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    • v.11 no.4
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    • pp.297-304
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    • 1998
  • The three analysis methods, EPA method 3050, the method offered by Ministry of Environment in Korea, and modified method corrected in this laboratory, were studied to investigate the effect of matrix on the analysis of inorganic priority pollutants. 7 inorganic priority pollutants(Ni, Cr, Cu, Zn, Pb, Cd, Hg) were spiked to the plating, leather, paper, electric, and dye sludges. Mean recovery of the elements except Hg was 95.5% when the procedure of EPA method was applied. However, recovery by the two other extraction methods showed 11.1% and 27.7%, respectively. Digestions were done by MDS (microwave digestion system) and $HNO_3+HClO_4$ methods. To study organic and inorganic matrix effect, samples were made by adding triethanol amine as a organic matrix and $FeCl_3{\cdot}6H_2O$+$AlCl_3{\cdot}6H_2O$ as a inorganic matrix, respectively. The extracts were analyzed by AAS and HG-AAS. Mean recovery of the elements by the $HNO_3+HClO_4$ procedure, except Hg, gave better result than that of the MDS method. Mean recovery of elements was decreased when organic and inorganic matrices were added in the sludge samples. The procedure of MDS and $HNO_3+HClO_4$ digestion gave higher recoveries than that of direct analysis. In general, the results of the studies showed a significant matrix effect on the inorganic priority pollutants analysis in sludges.

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Development of Protocol for the Effective Detection of Feline Calicivirus as Norovirus Surrogate in Oyster and Lettuce (굴과 상추에서 노로바이러스의 대체모델 feline calicivirus의 효율적 검출법 개발)

  • Lee, Soo-Yeon;Jang, Keum-Il;Woo, Gun-Jo;Kwak, Hyo-Sun;Kim, Kwang-Yup
    • Korean Journal of Food Science and Technology
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    • v.39 no.1
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    • pp.71-76
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    • 2007
  • Foodborne illness caused by Noroviruses (NVs) is increasing rapidly in Korea. This study developed an effective detection protocol for NVs found in contaminated oysters and lettuce through an investigation using the major steps of virus particle separation, concentration and RT-PCR. As a surrogate model for NVs, the cultivable feline calicivirus (FCV) that belongs to the same Caliciviridae family was used. Instead of using a time-consuming ultracentrifugation method, efficient methods based on solvent extraction and PEG precipitation procedure were applied. Direct homogenization of a 25g sample of whole oyster and lettuce in 175mL PBS provided the simplicity that would be needed in the actual field of food product examination. The overnight PEG precipitation step at $4^{\circ}C$ was reduced to 3 h by placing the reaction tube in ice and by adjusting the PEG concentrations. The application of the use of chloroform and 0.2 ${\mu}m$ syringe filtration together showed a better detection efficiency than the use of chloroform alone in removing PCR inhibitors for both oyster and lettuce samples. Also, dilution of the extracted RNA solution before PCR provided increased sensitivity. The improved detection protocol developed in this study could be efficiently applied to detect FCV and most likely NVs from oysters and lettuce.

Influence of the Cr-Carbides on the Mechanical Characteristics during Isothermal Heat-Treatment of the Mod.9Cr-1Mo Steel (Mod.9Cr-1Mo강의 항온변태시 기계적 특성변화에 미치는 Cr탄화물의 영향)

  • Hur, Sung-Kang;Lee, Jae-Hyun;Gu, Ji-Ho;Shin, Kee-Sam;He, Yinsheng;Shin, Jong-Ho
    • Korean Journal of Materials Research
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    • v.22 no.1
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    • pp.46-53
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    • 2012
  • In this study, mechanical tests and microstructural analyses including TEM analyses with EDX of precipitates in modified 9Cr-1Mo steel were carried out to determine the cause of embrittlement observed after heat-treatment, which limits the usage of the alloy for power plants. Mod. 9Cr-1Mo steel specimens at austenite temperature were quenched to the molten salt baths at $760^{\circ}C$ and $700^{\circ}C$, in which the specimens were kept for 10 min ~ 10 hr with subsequent air-cooling. Impact tests showed that the impact value dropped abruptly when the specimens were kept longer than 30 min at $\sim760^{\circ}C$ reaching to minima in about 1 hr, and then increasing at further retention. The tensile strength of the specimens reached the minimum value without much change afterward, whereas the values of elongation showed the same trend as that of the impact value. The isothermally heat-treated steel at $700^{\circ}C$ also showed a minimum impact value in about 1 hr. These results suggest that the isothermal heattreatment at 760 and $700^{\circ}C$ for about 1 hr induces temporal embrittlement in Mod. 9Cr-1Mo steel. The microstructural examination of all the specimens with extraction replica of the carbides revealed that the specimens with temporal embrittlement had $Cr_2C$, indicating that the cause of the embrittlement was the precipitation of the $Cr_2C$. In addition, TEM/EDX results showed that the Fe/Cr ratio was 0.033 to 0.055 for $Cr_2C$, whereas it was 0.48 to 0.75 for $Cr_{23}C_6$, making the distinction of the $Cr_2C$ and $Cr_{23}C_6$ possible even without direct electron diffraction analyses.

ZnO nanostructures for e-paper and field emission display applications

  • Sun, X.W.
    • 한국정보디스플레이학회:학술대회논문집
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    • 2008.10a
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    • pp.993-994
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    • 2008
  • Electrochromic (EC) devices are capable of reversibly changing their optical properties upon charge injection and extraction induced by the external voltage. The characteristics of the EC device, such as low power consumption, high coloration efficiency, and memory effects under open circuit status, make them suitable for use in a variety of applications including smart windows and electronic papers. Coloration due to reduction or oxidation of redox chromophores can be used for EC devices (e-paper), but the switching time is slow (second level). Recently, with increasing demand for the low cost, lightweight flat panel display with paper-like readability (electronic paper), an EC display technology based on dye-modified $TiO_2$ nanoparticle electrode was developed. A well known organic dye molecule, viologen, was adsorbed on the surface of a mesoporous $TiO_2$ nanoparticle film to form the EC electrode. On the other hand, ZnO is a wide bandgap II-VI semiconductor which has been applied in many fields such as UV lasers, field effect transistors and transparent conductors. The bandgap of the bulk ZnO is about 3.37 eV, which is close to that of the $TiO_2$ (3.4 eV). As a traditional transparent conductor, ZnO has excellent electron transport properties, even in ZnO nanoparticle films. In the past few years, one-dimension (1D) nanostructures of ZnO have attracted extensive research interest. In particular, 1D ZnO nanowires renders much better electron transportation capability by providing a direct conduction path for electron transport and greatly reducing the number of grain boundaries. These unique advantages make ZnO nanowires a promising matrix electrode for EC dye molecule loading. ZnO nanowires grow vertically from the substrate and form a dense array (Fig. 1). The ZnO nanowires show regular hexagonal cross section and the average diameter of the ZnO nanowires is about 100 nm. The cross-section image of the ZnO nanowires array (Fig. 1) indicates that the length of the ZnO nanowires is about $6\;{\mu}m$. From one on/off cycle of the ZnO EC cell (Fig. 2). We can see that, the switching time of a ZnO nanowire electrode EC cell with an active area of $1\;{\times}\;1\;cm^2$ is 170 ms and 142 ms for coloration and bleaching, respectively. The coloration and bleaching time is faster compared to the $TiO_2$ mesoporous EC devices with both coloration and bleaching time of about 250 ms for a device with an active area of $2.5\;cm^2$. With further optimization, it is possible that the response time can reach ten(s) of millisecond, i.e. capable of displaying video. Fig. 3 shows a prototype with two different transmittance states. It can be seen that good contrast was obtained. The retention was at least a few hours for these prototypes. Being an oxide, ZnO is oxidation resistant, i.e. it is more durable for field emission cathode. ZnO nanotetropods were also applied to realize the first prototype triode field emission device, making use of scattered surface-conduction electrons for field emission (Fig. 4). The device has a high efficiency (field emitted electron to total electron ratio) of about 60%. With this high efficiency, we were able to fabricate some prototype displays (Fig. 5 showing some alphanumerical symbols). ZnO tetrapods have four legs, which guarantees that there is one leg always pointing upward, even using screen printing method to fabricate the cathode.

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Guided tissue regeneration using barrier membranes on the dehiscence defects adjacent to the dental implants (치과용 임플란트 주위 열손 결손에 대한 차폐막의 유도조직재생에 관한 연구)

  • Lee, Dong-Ho;Choi, Sang-Mook
    • Journal of Periodontal and Implant Science
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    • v.25 no.2
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    • pp.301-320
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    • 1995
  • The purpose of this study was to evaluate a new biodegradable membrane - atelocollagen as a guided tissue regeneration barrier on the dehiscence defects adjacent to the dental implants. 3 beagle dogs were selected for this study and all the mandibular premolars($P_1,P_2,P_3&P_4$) were extracted. Twelve weeks after the extraction, the edentulous ridges were formed to be placed the titanium plasma-sprayed IMZ implants. Four implant osteotomies were performed on each side of the mandible. The osteotomies were placed facially in the edentulous ridges to approximate an actual dehiscence defect as closely as possible, The standardized dehiscence defects were created 3 mm in width and 4 mm in height by osteotomy. A total 24 implants were placed. e-PTFE, ateloco11agen and $Collatape^{(R)}$ were placed to cover the defects and the one defect served as a control, not covered any membrane. By random selection, three dogs were sacrificed at 2 weeks, 4weeks and 8 weeks after fixation with 3% glutaraldehyde. A week before sacrificing, 8-week dog was infused intravenously with oxy-tetracycline 30mg/kg. The left mandibular blocks were used for full decalcified histologic preparation and the right mandibular blocks were selected for undeca1cified preparation, At 2 weeks, the regenerated bone of e-PTFE and atelocollagen groups appeared to be more dense than other groups and the percentage of bone defect fill was highest for e-PTFE and follwed by ateloco1lagen group. However, the $Collatape^{(R)}$ and control groups showed a little new bone formation. $Collatape^{(R)}$ was almost degraded within 2 weeks. At 4 weeks, the regenerated new bone were much greater and denser than at 2 weeks for e-PTFE and ateloco11agen group. Although a part of atelocollagen bagan to be degraded at the margin and surrounded by foreign body giant cells related to foreign body reaction, it was generally intact and the regenerated new bone was shown much more than at 2 weeks. The amount of new bone in $Collatape^{(R)}$ and control groups at 4 weeks were similar to that of 2 weeks group. At 8 weeks, the regenerated bone was matured and observed along the implant fixture. Direct new bone formation and calcium deposits beneath the e-PTFE were observed. No further bone growth was seen in the $Collatape^{(R)}$ and control groups. In reflected fluoromicrcocopic observation, the osteogenic activity was pronounced between e-PTFE membrane and the old bone. High osteogenic activity was also observed in atelocol1agen group. This study suggested that the ateloco11agen as well as e-PTFE could be used for guided tissue regeneration on dehiscence defects adjacent to the dental implants. But the $Collatape^{(R)}$ was completely resorbed within 2 weeks and was not a suitable membrane for guided bone regeneration.

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