• The Plant Pathology Journal
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• 제33권5호
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• pp.508-513
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• 2017

The complete genome sequence of a Slovak SL-1 isolate of Tomato mosaic virus (ToMV) was determined from the next generation sequencing (NGS) data, further confirming a limited sequence divergence in this tobamovirus species. Tomato genotypes Monalbo, Mobaci and Moperou, respectively carrying the susceptible tm-2 allele or the Tm-1 and Tm-2 resistant alleles, were tested for their susceptibility to ToMV SL-1. Although the three tomato genotypes accumulated ToMV SL-1 to similar amounts as judged by semiquantitative DAS-ELISA, they showed variations in the rate of infection and symptomatology. Possible differences in the intra-isolate variability and polymorphism between viral populations propagating in these tomato genotypes were evaluated by analysis of the capsid protein (CP) encoding region. Irrespective of genotype infected, the intra-isolate haplotype structure showed the presence of the same highly dominant CP sequence and the low level of population diversity (0.08-0.19%). Our results suggest that ToMV CP encoding sequence is relatively stable in the viral population during its replication in vivo and provides further demonstration that RNA viruses may show high sequence stability, probably as a result of purifying selection.

• Journal of Microbiology and Biotechnology
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• 제25권4호
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• pp.511-520
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• 2015

Triclosan, the widely used biocide, specifically targets enoyl-acyl carrier protein reductase (ENR) in the bacterial fatty acid synthesis system. Although the fish pathogen Aeromonas salmonicida subsp. salmonicida exhibits triclosan resistance, the nature of this resistance has not been elucidated. Here, we aimed to characterize the triclosan resistance of A. salmonicida subsp. salmonicida causing furunculosis. The fosmid library of triclosan-resistant A. salmonicida subsp. salmonicida was constructed to select a fosmid clone showing triclosan resistance. With the fosmid clone showing triclosan resistance, a subsequent secondary library search resulted in the selection of subclone pTSR-1. DNA sequence analysis of pTSR-1 revealed the presence of a chromosomal-borne fabV-encoding ENR homolog. The ENR of A. salmonicida (FabVas) exhibited significant homology with previously known FabV, including the catalytic domain YX₍₈₎K. fabVas introduction into E. coli dramatically increased its resistance to triclosan. Heterologous expression of FabVas might functionally replace the triclosan-sensitive FabI in vivo to confer E. coli with triclosan resistance. A genome-wide search for fabVas homologs revealed the presence of an additional fabV gene (fabVas2) paralog in A. salmonicida strains and the fabVas orthologs from other gram-negative fish pathogens. Both of the potential FabV ENRs expressed similarly with or without triclosan supplement. This is the first report about the presence of two potential FabV ENRs in a single pathogenic bacterium. Our result suggests that triclosan-resistant ENRs are widely distributed in various bacteria in nature, and the wide use of this biocide can spread these triclosan-tolerant ENRs among fish pathogens and other pathogenic bacteria.

• BMB Reports
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• 제34권1호
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• pp.51-58
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• 2001

Attempts using in vitro and in vivo selection procedures have been made to search for hammerhead ribozymes that have higher activities than the wild-type ribozyme and also to determine whether other sequences might be possible in the catalytic core of the hammerhead ribozyme. Active sequences selected in the past conformed broadly to the consensus core sequence except at A9, and no sequences were associated with higher activity than that of the hammerhead with the consensus core, an indication that the consensus sequence derived from viruses and virusoids is probably the optimal sequence [Vaish et al. (1997) Biochemistry 36, 6495-6501]. Recently, during construction of ribozyme expression vectors, we isolated a mutant hammerhead ribozyme, with an insertion of G between A9 and G10.1, that appeared to show significant activity [Kawasaki et al. (1996) Nucleic Acids Res. 24, 3010-3016; Kawasaki et al. (1998) Nature 393, 284-289]. We, therefore, characterized kinetic properties of the G-inserted mutant ribozymes in terms of the NUX rule. We demonstrate that the NUX rule is basically applicable to the G-inserted ribozymes and, more importantly, one type of G-inserted ribozyme was very active with $k_{cat}$, value of $6.4\;min^{-1}$ in 50 mM Tris-HCl (pH 8.0) and 10 mM $MgCl_2$ at $37^{\circ}C$.

• Journal of Plant Biotechnology
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• 제30권4호
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• pp.307-313
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• 2003

파리지옥풀 first trap leaf에만 발현하는 유전자군을 탐색하기 위하여 기내배양 식물체와 포충능력을 가진 3년생 실생주을 이용하여 각각의 포충잎 (leaf base), 꽃조직 (flower tissue) 및 포충잎트랩 (first trap leaf)으로부터 분리한 RNA로 Fluorescent differential display (FDD)를 실시하였다. First trap leaf특이발현 유전자 15개를 screening하여 염기서열을 분석하였다. 분리된 DNA들은 protease inhibitor (Pl), myo-inositol-1-phosphate synthase 및 lipocalin-type prostaglandin D syn-thase 유전자들과 매우 유사하였다. 또한 Northern blot분석 결과, 이들 유전자들이 first trap leaf에 특이적으로 발현하고 있는 것을 확인하였다 FDD방법은 세포, 조직 및 기관에 특이적으로 발현하고 있는 유전자들을 선발하는데 매우 유용한 수단으로 사용될 수 있다.

• IMMUNE NETWORK
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• 제2권4호
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• pp.233-241
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• 2002

Background: Monoclonal antibodies (mAb) of rodent origin are produced with ease by hybridoma fusion technique, and have been successfully used as therapeutic reagents for humans after humanization by genetic engineering. However, utilization of these antibodies for therapeutic purpose has been limited by the fact that they act as immunogens in human body causing undesired side effects. So far, there have been several attempts to produce human mAbs for effective in vivo diagnostic or therapeutic reagents including the use of humanized xenomouse that is generated by mating knockout mice which lost Ig heavy and light chain genes by homologous recombination and transgenic mice having both human Ig heavy and light gene loci in their genome. Methods: Genomic DNA fragments of mouse Ig heavy and light chain were obtained from a mouse brain ${\lambda}$ genomic library by PCR screening and cloned into a targeting vector with ultimate goal of generating Ig knockout mouse. Results: Through PCR screening of the genomic library, three heavy chain and three light chain Ig gene fragments were identified, and restriction map of one of the heavy chain gene fragments was determined. Then heavy chain Ig gene fragments were subcloned into a targeting vector. The resulting construct was introduced into embryonic stem cells. Antibiotic selection of transfected cells is under the progress. Conclusion: Generation of xenomouse is particularly important in medical biotechnology. However, this goal is not easily achieved due to the technical difficulties as well as huge financial expenses. Although we are in the early stage of a long-term project, our results, at least, partially contribute the successful generation of humanized xenomouse in Korea.

• Imaging Science in Dentistry
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• 제52권2호
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• pp.133-140
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• 2022

Purpose: The aim of this review was to systematically analyze the available literature on the correlation between the gray values (GVs) of cone-beam computed tomography (CBCT) and the Hounsfield units (HUs) of computed tomography (CT) for assessing bone mineral density. Materials and Methods: A literature search was carried out in PubMed, Cochrane Library, Google Scholar, Scopus, and LILACS for studies published through September 2021. In vitro, in vivo, and animal studies that analyzed the correlations GVs of CBCT and HUs of CT were included in this review. The review was prepared according to the PRISMA checklist for systematic reviews, and the risk of bias was assessed using the Quality Assessment of Diagnostic Accuracy Studies tool. A quantitative analysis was performed using a fixed-effects model. Results: The literature search identified a total of 5,955 studies, of which 14 studies were included for the qualitative analysis and 2 studies for the quantitative analysis. A positive correlation was observed between the GVs of CBCT and HUs of CT. Out of the 14 studies, 100% had low risks of bias for the domains of patient selection, index test, and reference standards, while 95% of studies had a low risk of bias for the domain of flow and timing. The fixed-effects meta-analysis performed for Pearson correlation coefficients between CBCT and CT showed a moderate positive correlation (r=0.669; 95% CI, 0.388 to 0.836; P<0.05). Conclusion: The available evidence showed a positive correlation between the GVs of CBCT and HUs of CT.

• 대한한방부인과학회지
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• 제26권4호
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• pp.191-203
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• 2013

Purpose: This review is to figure out evidence that suggest effectiveness of Korean Medicine treatments against cervical dysplasia. Methods: Studies on cervical cancer and cervical dysplasia were searched through 6 databases: Korean Studies Information Service System(KISS), National Discovery for Science Leaders (NDSL), Korea Institute of Science and Technology Information (Korean TK), Oriental Medicine Advanced Searching Integrated System (OASIS), the Journal of Korean Medicine, and the Journal of Korean Obstetrics & Gynecology. After that, the articles were extracted with reference point of Korean Traditional Medicine. Results: 37 articles were included lastly according to selection criteria. 3 of them were case reports on cervical dysplasia, and 34 were in-vitro studies on Human Papilloma Virus (HPV) positive cancer cell. In case reports, acupuncture, moxibustion, medical herbs and pharmacoacupuncture were used for treatments of cervical dysplasia with about 3 months. Experimental studies on cervical cancer cell showed that several herbs function with clear heat, eliminate stasis (淸熱解毒, 化瘀消腫) have anti-cancer effects inducing apoptosis. Conclusions: The results of articles are not enough to use in practice. Therefore, we indicates more advanced research methodology as follows: development of Korean Medicine treatment protocol with oral and external, in-vivo experimental study, and evaluation immunity index.

• 대한방사성의약품학회지
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• 제3권2호
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• pp.103-112
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• 2017

The combination of nanoparticle with radioisotope could give the in vivo information with high sensitivity and specificity. However, radioisotope labeling of nanoparticle is very difficult and radioisotopes have different physicochemical properties, so the radioisotope selection of nanoparticle should be carefully considered. $^{18}F$ was first option to be considered for labeling of nanoparticle. For the labeling of $^{18}F$ with nanoparticle, Prosthetic group is widely used. Iodine, another radioactive halogen, is often used. Since radioiodine isotopes are various, they can be used for different imaging technique or therapy in the same labeling procedures. $^{99m}Tc$ can easily be obtained as pertechnatate ($^{99m}{TcO_4}^-$) by commercial generator. Ionic $^{68}Ga$ (III) in dilute HCl solution is also obtained by generator system, but $^{68}Ga$ can be substituted for $^{67}Ga$ because of the short half-life (67.8 min). $^{64}Cu$ emits not only positron but also ${\beta}-particle$. Therefore $^{64}Cu$ can be used for imaging and therapy at the same time. These radioactive metals can be labeled with nanoparticle using the bifunctional chelator. $^{89}Zr$ has longer half-life (78.4 h) and is used for the longer imaging time. Unlike different metals, $^{89}Zr$ should use the other chelate such as DFO, 3,4,3-(LI-1,2-HOPO) or DFOB.

• Applied Biological Chemistry
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• 제47권1호
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• pp.17-21
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• 2004

경작지 근권토양에서 식물생장촉진물질인 auxin라 항진균물질을 동시에 생산하는 농업용 미생물제제로 사용할 PGPR 균주를 선발하고 동정하였다. 경북 경산 일대의 다수확 밭토양의 근권토양에서 균주를 선발하고 그 중 auxin 생산성이 높은 2균주 K36, N21를 분리하였다. 이들 균주를 동정한 결과 K36은 Pantoea agglomerans이고 N21은 Pseudomonans fulva로 동정되었다. 선발된 균주의 식물생장촉진능을 in vivo 녹두발아촉진검사법으로 확인하였으며, 식물병원진균에 대한 길항력을 확인하였다. 배양조건에 따른 auxin 생산성을 조사한 결과 K36 균주는 $pH\;7.5,\;35^{\circ}C$, 37시간에서 최대로 나타났고, N21균주는 $pH\;7.5,\;35^{\circ}C$, 20시간에서 최대값을 나타내었다.

• The Journal of Advanced Prosthodontics
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• 제5권4호
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• pp.382-387
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• 2013

PURPOSE. Although several surface treatments have been recently investigated both under in vitro and in vivo conditions, controversy still exists regarding the selection of the most appropriate zirconia surface pre-treatment. The purpose of this study was to evaluate the effect of alumina (Al) and aluminium nitride (AlN) coating on the shear bond strength of adhesive resin cement to zirconia core. MATERIALS AND METHODS. Fifty zirconia core discs were divided into 5 groups; air particle abrasion with 50 ${\mu}m$ aluminum oxide particles ($Al_2O_3$), polishing + Al coating, polishing + AlN coating, air particle abrasion with 50 ${\mu}m$ $Al_2O_3$ + Al coating and air particle abrasion with 50 ${\mu}m$ $Al_2O_3$ + AlN coating. Composite resin discs were cemented to each of specimens. Shear bond strength (MPa) was measured using a universal testing machine. The effects of the surface preparations on each specimen were examined with scanning electron microscope (SEM). Data were statistically analyzed by one-way ANOVA (${\alpha}$=.05). RESULTS. The highest bond strengths were obtained by air abrasion with 50 ${\mu}m$ $Al_2O_3$, the lowest bond strengths were obtained in polishing + Al coating group (P<.05). CONCLUSION. Al and AlN coatings using the reactive magnetron sputtering technique were found to be ineffective to increase the bond strength of adhesive resin cement to zirconia core.