• Title/Summary/Keyword: Random-priming

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Isolation of a Variant Strain of Pleurotus eryngii and the Development of Specific DNA Markers to Identify the Variant Strain

  • Lee, Hyun-Jun;Kim, Sang-Woo;Ryu, Jae-San;Lee, Chang-Yun;Ro, Hyeon-Su
    • Mycobiology
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    • v.42 no.1
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    • pp.46-51
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    • 2014
  • A degenerated strain of Pleurotus eryngii KNR2312 was isolated from a commercial farm. Random amplified polymorphic DNA analysis performed on the genomic DNA of the normal and degenerated strains of this species revealed differences in the DNA banding pattern. A unique DNA fragment (1.7 kbp), which appeared only in the degenerated strain, was isolated and sequenced. Comparing this sequence with the KNR2312 genomic sequence showed that the sequence of the degenerated strain comprised three DNA regions that originated from nine distinct scaffolds of the genomic sequence, suggesting that chromosome-level changes had occurred in the degenerated strain. Using the unique sequence, three sets of PCR primers were designed that targeted the full length, the 5' half, and the 3' half of the DNA. The primer sets P2-1 and P2-2 yielded 1.76 and 0.97 kbp PCR products, respectively, only in the case of the degenerated strain, whereas P2-3 generated a 0.8 kbp product in both the normal and the degenerated strains because its target region was intact in the normal strain as well. In the case of the P2-1 and P2-2 sets, the priming regions of the forward and reverse primers were located at distinct genomic scaffolds in the normal strain. These two primer sets specifically detected the degenerate strain of KNR2312 isolated from various mushrooms including 10 different strains of P. eryngii, four strains of P. ostreatus, and 11 other wild mushrooms.

Type of Classification Criterion and Characteristic of Classification Strategy That Appear in Pre-Service Elementary Teachers' Classification Activity (예비 초등 교사들의 분류 활동에서 나타난 분류 기준의 유형과 분류 전략의 특징)

  • Yang, Il-Ho;Choi, Hyun-Dong
    • Journal of Korean Elementary Science Education
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    • v.27 no.1
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    • pp.9-22
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    • 2008
  • The purpose of this study was to investigate the type of classification criterion and the characteristic of classification strategy that appear in pre-service elementary teachers' classification activity. The 4 tasks were developed for classification activity; button as a real things that attribute is prominent, shell as a real things that attribute is less prominent, snow flake as a picture cards that attribute is prominent, and galaxy as a picture cards that attribute is less prominent. The 5 college students who major in elementary education were selected. Data were collected by interview with participants, participants' classification recording paper, investigator's observation of participants' action observation, and videotaped that record participants' subject classification process. Result proved in this study is as following. First, pre-service elementary teachers used 4 qualitative classification criterion of feature, random field, image and secondary property, and used 2 dimension classification criterion of space and quantity. They used single quality classification criterion or combining dimension classification criterion in classification activity. Second, pre-service elementary teachers have classification strategy that apply each various classification criterion, and also classification strategy are different according to subject, but discussed that "anchor" and "priming effect" are important for effective classification. Result of this study is expected to contribute classification research and classification teaching program development.

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Identification of Differentially Expressed Genes by Gabapentin in Cultured Dorsal Root Ganglion in a Rat Neuropathic Pain Model

  • Heo, Ji Hye;Lee, Seung Ha;Chang, Kyung Ha;Han, Eun Hye;Lee, Seung Gwan;Choi, Dal Woong;Kim, Suhng Wook
    • Biomolecules & Therapeutics
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    • v.21 no.2
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    • pp.126-131
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    • 2013
  • Neuropathic pain is a chronic pain disorder caused by nervous system lesions as a direct consequence of a lesion or by disease of the portions of the nervous system that normally signal pain. The spinal nerve ligation (SNL) model in rats that reflect some components of clinical pain have played a crucial role in the understanding of neuropathic pain. To investigate the direct effects of gabapentin on differential gene expression in cultured dorsal root ganglion (DRG) cells of SNL model rats, we performed a differential display reverse transcription-polymerase chain reaction analysis with random priming approach using annealing control primer. Genes encoding metallothionein 1a, transforming growth factor-${\beta}1$ and palmitoyl-protein thioesterase-2 were up-regulated in gabapentin-treated DRG cells of SNL model rats. The functional roles of these differentially expressed genes were previously suggested as neuroprotective genes. Further study of these genes is expected to reveal potential targets of gabapentin.

Identification of Differentially Expressed Genes in Human Mesenchymal Stem Cell-Derived Neurons

  • Heo, Ji-Hye;Cho, Kyung-Jin;Choi, Dal-Woong;Kim, Suhng-Wook
    • Toxicological Research
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    • v.26 no.1
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    • pp.15-19
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    • 2010
  • Mesenchymal stem cells (MSCs) have greater potential for immediate clinical and toxicological applications, due to their ability to self-renew, proliferate, and differentiate into a variety of cell types. To identify novel candidate genes that were specifically expressed during transdifferentiation of human MSCs to neuronal cells, we performed a differential expression analysis with random priming approach using annealing control primer-based differential display reverse transcription-polymerase chain reaction approach. We identified genes for acyl-CoA thioesterase, tissue inhibitor of metalloproteinases-1, brain glycogen phosphorylase, ubiquitin C-terminal hydrolase and aldehyde reductase were up-regualted, whereas genes for transgelin and heparan sulfate proteoglycan were down-regulated in MSC-derived neurons. These differentially expressed genes may have potential role in regulation of neurogenesis. This study could be applied to environmental toxicology in the field of testing the toxicity of a chemical or a physical agent.

Possibility of Using DNA Chip Technology for Diagnosis of Human Papillomavirus

  • Liu, Cui-Hua;Ma, Wen-Li;Shi, Rong;Ou, Yang-Qian;Zhang, Bao;Zheng, Wen-Ling
    • BMB Reports
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    • v.36 no.4
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    • pp.349-353
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    • 2003
  • To explore the application of DNA chip technology for the detection and typing of Human Papillomavirus (HPV), the HPV6, 11, 16 and 18 gene fragments were isolated and printed onto aminosilane-coated glass slides by a PixSys 5500 microarrayer as probes to prepare the HPV gene chips. HPV samples, after being labeled with fluorescent dye by restriction display PCR (RD-PCR) technology, were hybridized with the microarray, which was followed by scanning and analysis. The experimental condition for preparing the HPV gene chips was investigated, and the possibility of HPV genotyping using gene chips was discussed. The technique that was established in this study for preparing HPV gene chips is practical. The results of the present study demonstrated the versatility and inspiring prospect of using this technology to detect and genotype HPV.

Changes of Total and Ionized Calcium following Cardiopulmonary Bypass (심폐관류에 따른 혈청칼슘의 변동)

  • 전상훈
    • Journal of Chest Surgery
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    • v.21 no.2
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    • pp.240-245
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    • 1988
  • This study was prospectively planned to realize the reduction of calcium ion in serum along with the cardiopulmonary bypass[CPB], to find out the cause of the reduction, and to verify the justification of the classical methods of calcium replacement. Nine patients with various open heart surgeries by CPB in 1987 wee selected at random. Calcium chloride was added as follows:: For each unit of ACD blood transfusion, 600mg of calcium chloride was added. In case of massive transfusion, 600 mg of calcium chloride was injected every 2 or 3 units of transfusion. On occasions such as weaning from CPB, or following defibrillation, or hypotension, weak myocardial contractility of the heart, calcium chloride was needed in an amount of 10 mg / kg. In ICU, calcium chloride was limited to use in low serum level or in emergency use. Total calcium decreased early bypass and progressively increased above the preoperative value during late bypass and three hours thereafter, Ionized calcium increased during late bypass and three hours following. Total and ionized calcium depicted similar patterns of change during open heart surgery. Decrease of the calcium at the early bypass was thought from reduction of total protein and alkalosis during bypass. Meanwhile, increase of both calciums during the end of surgery was presumably attributable to addition of calcium chloride in priming solution, injections of calcium chloride in the process of termination of bypass. We conclude that enough calcium was replaced by the classical methods of calcium supplement.

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EFFECTS OF MOISTENING OF ETCHED DENTIN AND ENAMEL SURFACES ON BOND STRENGTH (산 표면처리후 상아질과 법랑질 표면의 습윤이 결합강도에 미치는 영향)

  • Lee, Kwang-Won;Park, Soo-Joung;Son, Ho-Hyun
    • Restorative Dentistry and Endodontics
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    • v.20 no.1
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    • pp.328-341
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    • 1995
  • I. Shear Bond Strength to Air-dried and Remoistened Dentin.. The effect of air-drying and remoistening of acid-conditioned dentin before priming with the primer of All-Bond 2(BISCO. INC., U. S. A.) on shear bond strength(SBS) was investigated. Ninty freshly extracted sound human molars were divided at random into 9 groups of 10 teeth each. SBSs were meaured for acid-conditioned and non-conditioned dentin to which the primer and bonding agent of All-Bond 2 and composite resin(Z-100, 3M Dental Products, U. S. A.) were applied. The following values(Mean${\pm}$ SD, MPa) were obtained for the groups conditioned with 10% phosphoric acid for 15 seconds: Group l(blot dried) $6.7{\pm}4.1$ ; Group 2(10 seconds dried) $16.1{\pm}5.3$ ; Group 3(20 seconds dried) $15.4{\pm}4.8$ ; Group 4(30 seconds dried) $15.2{\pm}6.3$ ; Group 5(10 seconds dried/remoistened) $26.4{\pm}2.6$ ; Group 6(20 seconds dired/remositened) $22.2{\pm}2.7$ ; Group 7(30 seconds dried/remoistened) $21.5{\pm}4.1$. For the non-conditioned groups the values were: Group 8 (blot dried) $13.3{\pm}2.6$ ; Group 9(10 seconds dried) $12.9{\pm}3.5$. The data were analyzed using ANOVA. In the acid-conditioned groups, mean values of SBS for the air-dried specimens(Grps. 2, 3 and 4) and the 20 and 30 seconds dried/remoistened specimens (Grps. 6 and 7) were significantly lower than that of blot dried specimens.(p<0.05) The value for 10 seconds dried/remoistened specimens (Grp. 5), however, was not statistically different compared to that of blot dried specimens.(p>0.05) In the non-conditined groups, there was no statistical difference between blot dried and 10 seconds dried specimens.(p>0.05) The results suggest that the acid-conditioned dentin surface is more vulnerable to dentin bonding when it is air-dried or even remoistened after long period of drying. II. Shear bond stengh to the moistened and primed enamel. The effect of moistening and priming of enamel compared to the air-drying of enamel on the shear bond strength of enamel bonding agent was investigated. The experiment was divided into 4 groups each containing 10 caries-free maxillary incisor teeth. Shear bond strength values were measured for the primed and non-primed enamel to which All-Bond 2 and Z-100 were applied. The following values(MPa) were obtained for the primed groups pretreated with 32 % phosphoric acid for 15 seconds. : Group 1 (10 seconds dried) $29.8{\pm}2.2$ ; Group 2(moistened) $26.8{\pm}5.4$. For the non-primed groups the values were: Group 3(10 seconds dried/primed) $27.6{\pm}5.0$ ; Group 4(mostened/primed) $28.2{\pm}3.5$. The data were subjected to statistical analysis using ANOVA. The results showed that mean shear bond strengths among the experimental groups were not statistically different. (p>0.05) Conclusively, It is suggested that the bonding ability to enamel is not decreased by the moistening and priming of the enamel.

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The Expression of Cytokines in the Airways from Patients with Bronchial Asthma (천식환자의 기도내 Cytokines 표현에 대한 연구)

  • Uh, Soo-Taek;Jeong, Seong-Whan;Lee, Sang-Moo;Kim, Hyeon-Tae;Kim, Yong-Hoon;Park, Choon-Sik
    • Tuberculosis and Respiratory Diseases
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    • v.42 no.1
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    • pp.1-10
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    • 1995
  • Background: It has been well known that bronchial asthma is a chronic inflammatory disorder. The "activation" of lymphocytes has a significant role in the pathogenesis of bronchial asthma. Among these lymphocytes, TH2-like rather than TH1-like lymphoytes are activated in the bronchial tissues from patients with atopic bronchial asthma. However, the difference of cytokines expression is not well documented between the atopic normal subjects and atopic asthmatics. Methods: Bronchial tissues were obtained from the tweleve atopic and non-atpoic asthmatics and tweleve atopic and non-atopic healthy subjects for in stiu hybridizatin of IL-2, IL-4, IL-5, and INF-$\gamma$. The probe of cytokines were tagged with digoxigenin by random priming method. Results: The infiltration of many inflammatory cells on submucosa and denuded epithelium were observed in the bronchial tissue from patients with bronchial asthma. The RNase-treated bronchial tissues did not have the brown signal on the tissue, but, RNasc-untreated bronchial tissues had the positive brown signal on the inflammatory cells under the basement membrane. The IL-2 positive signals were detected in 2 cases, IFN-$\gamma$ in 1 casc, IL-4 in 2 cases, IL-5 in 2 cases among 6 non-atopic healthy subjects. The atopic healthy subjects showed 1 case of positive signal of IL-2 and IFN-$\gamma$, but did not show any signals of IL-4 and IL-5. The positive signals of IL-2 were detected in 4 cases among 6 atopic and 6 non-atopic asthmatics, 2 cases and 1 case of IFN-$\gamma$ respectively, 4 cases and 3 cases of IL-4 respectively, 4 cases and 3 cases of IL-5 respectively. Conclusion: The lymphocytes were activated in the bronchus of asthmatics. Among lymphocytes, TH2-like lymphocytes may be involved in the pathogenesis of bronchial asthma. However, futher study with immunohistochemical stain may be necessary for defining the source of cytokines, because of TH2-like lymphocytes were also activated in some atopic healthy subjects.

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ELECTRON MICROSCOPIC STUDY OF RESIN-DENTIN HYBRID ZONE PRODUCED BY THE MOISTENING OF ACID CONDITIONED DENTIN SURFACE (산 표면처리 후 상아질 표면의 습윤이 하이브리드층 형성에 관한 전자현미경적 연구)

  • Lee, Kwang-Won;Lee, Myung-Jong
    • Restorative Dentistry and Endodontics
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    • v.20 no.2
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    • pp.463-486
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    • 1995
  • The effect of moistening and air-drying of acid-conditioned dentin before priming on the formation of resin-dentin hybrid zone was investigated, Freshly extracted human molars were used and divided at random into 5 groups, Groups 1 - 3 consisted of specimens conditioned with 10 % phosphoric acid for 20 seconds; Group 1 served as a control in which the conditioned dentin was simply blot-dried with a damp facial tissue; Group 2 was air dried for 30 seconds ; Group 3 was air dried for 30 seconds and immediately remoistened for 10 seconds with air-water syringe. and then the specimen was blot-dried with a damp facial tissue. Groups 4-5 were not acid conditioned ; In group 4, the smear layer on the dentin was blot dried before primer placement; Group 5 was air dried only for 30 seconds, The acetone-based primer and bonding agent of All Bond 2 (Bisco. Inc., USA) and composite resin (Z-100, 3M Dental products, USA) were applied for acid conditioned dentin and non-conditioned dentin. The morphologic ultrastructure of resin-dentin hybrid zone was examined by the use of SEM and TEM. and the existence of inorganic material and analysis of Ca/P weight-percent ratio in the resin-dentin hybrid zone were revealed by the EDAX, The results were as follows : 1. In the moistened specimens from acid-conditioned groups, the resin penetrated about 3-$4{\mu}m$ into dentin and the denatured collagen smear layer was not present at the surface. The resin tag was formed to a thickeness of 3-$4{\mu}m$ at the upper part of dentinal tubule and compactively connected to each other by means of many lateral branching. 2. In the air-dried specimens from acid-conditioned groups, the resin penetrated about 2.0-$2.5\;{\mu}m$ into dentin and an upper thin black layer to a thickness of 30-35nm was identified between adhesive resin and demineralized collagen layer. The resin tag to have a diameter of $2.5{\mu}m$ was formed at the upper part of dentinal tubule. However the funnel shape of the tag was not notable compared to the moistened specimens. 3. In the remoistened specimens from acid conditioned groups, the resin penetrated about 2.0-$2.5{\mu}m$ into dentin and an upper black layer was not present. The resin tag at the upper part of dentinal tubule was formed less than $2{\mu}m$ and was weakly connected to each other by means of few lateral branching. 4. In the non-conditioned groups, the smear layer was formed to a thickness of $0.5{\mu}m$ at dentin surface. However, the resin-dentin hybrid zone was not identified by TEM. The evidence of resin penetration into intertubular and intratubular dentin did not show. 5. All the acid-conditioned groups showed that the detected calcium and phosphorus weight percent ratios at the $2{\mu}m$ upper portion from the resin-dentin interface into the resin were much higher than that at the $2{\mu}m$ lower portion from the resin-dentin interface to dentin. (P<0.01).

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