• Restorative Dentistry and Endodontics
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• v.36 no.6
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• pp.498-505
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• 2011

Objectives: The purpose of this in vivo study was to investigate the microbial diversity in symptomatic and asymptomatic canals with primary endodontic infections by using GS FLX Titanium pyrosequencing. Materials and Methods: Sequencing was performed on 6 teeth (symptomatic, n = 3; asymptomatic, n = 3) with primary endodontic infections. Amplicons from hypervariable region of the small-subunit ribosomal RNA gene were generated by polymerized chain reaction (PCR), and sequenced by means of the GS FLX Titanium pyrosequencing. Results: On average, 10,639 and 45,455 16S rRNA sequences for asymptomatic and symptomatic teeth were obtained, respectively. Based on Ribosomal Database Project Classifier analysis, pyrosequencing identified the 141 bacterial genera in 13 phyla. The vast majority of sequences belonged to one of the seven phyla: Actinobacteria, Bacteroidetes, Firmicutes, Fusobacteria, Proteobacteria, Spirochetes, and Synergistetes. In genus level, Pyramidobacter, Streptococcus, and Leptotrichia constituted about 50% of microbial profile in asymptomatic teeth, whereas Neisseria, Propionibacterium, and Tessaracoccus were frequently found in symptomatic teeth (69%). Grouping the sequences in operational taxonomic units (3%) yielded 450 and 1,997 species level phylotypes in asymptomatic and symptomatic teeth, respectively. The total bacteria counts were significantly higher in symptomatic teeth than that of asymptomatic teeth (p < 0.05). Conclusions: GS FLX Titanium pyrosequencing could reveal a previously unidentified high bacterial diversity in primary endodontic infections.

• Journal of Dental Rehabilitation and Applied Science
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• v.30 no.2
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• pp.121-130
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• 2014

Purpose: To assess the surface profile of dentinal wall, dentin chips and smear layer during the canal shaping with rotary (ProTaper) and ProFile and reciprocating (WaveOne) nickel-titanium file. Materials and Methods: Sixty human extracted mandibular premolars and incisors with single canals were randomly selected. Three experimental groups (n = 20) were instrumented with ProTaper (F2), ProFile (25/.06), WaveOne (25/.08) with irrigation of 2.5% NaOCl. The dentin chips were collected from flute of file during each canal preparation. After canal preparation, roots were grinded and each group was divided into two subgroups (n = 10) for surface profile and smear layer of dentinal wall of shaped root canal. Each specimen was observed under scanning electron microscope for evaluating size of dentin chips, root canal surface recessions and smear layer. Scores of Smear layer were statistically analyzed using Kruskal Wallis test and Mann Whitney test at P = 0.05 level. Results: The size of dentin chips from ProFile, ProTaper and WaveOne was up to $7{\mu}m$, $6.5{\mu}m$, and$4{\mu}m$, respectively. In the surface profile, the width of surface irregularity was measured and Profile, ProTaper and WaveOne was up to $150{\mu}m$, $70{\mu}m$, and $80{\mu}m$, respectively. Completely cleaned root canals were not found. In the middle and apical third of the canals, WaveOne group showed higher smear layer score than ProFile and ProTaper groups (P < 0.05). Conclusion: Within limits of this study, reciprocating motion WaveOne group was not significant difference of shaping ability with the full-sequence ProFile and ProTaper systems except canal clearness of middle and apical third of root canal. When using WaveOne to shaping root canal, thorough root canal irrigation is recommended.

• Progress in Medical Physics
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• v.13 no.3
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• pp.120-128
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• 2002

To investigate differences between the metabolic ratios of normal controls and brain tumors such as astrocytomas and glioblastoma multiforme (GM) by proton MR spectroscopy (MRS) at 37 high field system. Using 3T MRI/MRS system, localized water-suppressed single-voxel technique in patients with brain tumors was employed to evaluate spectra with peaks of N-acetyl aspartate (NAA), choline-containing compounds (Cho), creatine/phosphocreatine (Cr) and lactate. On the basis of Cr, these peak areas were quantificated as a relative ratio. The variation of metabolites measurements of the designated region in 10 normal volunteers was less than 10%. Normal ranges of NAA/Cr and Cho/Cr ratios were 1.67$\pm$018 and 1.16$\pm$0.15, respectively. NAA/Cr ratio of all tumor tissues was significantly lower than that of the normal tissues (P=0.005). Cho/Cr ratio of glioblastoma multiforme was significantly higher than that of astrocytomas (P=0.001). Lactate was observed in all tumor cases. The present study demonstrated that the neuronal degradation or loss was observed in all tumor tissues. Higher grade of brain tumors was correlated with higher Cho/Cr ratio, indicating a significant dependence of Cho levels on malignancy of gliomas. This results suggest that clinical proton MR spectroscopy could be useful to predict tumor malignancy.

• Journal of Life Science
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• v.30 no.8
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• pp.701-707
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• 2020

Gene editing technology using the clustered regularly interspaced short palindromic repeat (CRISPR) and the CRISPR associated protein (Cas)9 has been highly anticipated in developing breeding techniques. In this study, we discuss gene scissors as a tool for silkworm molecular breeding through analysis of Bombyx mori Kynurenine 3-Monooxygenase (BmKMO) gene editing using the CRISPR/Cas9 system and analysis of generational transmission through mutagenesis and selective crossing. The nucleotide sequence of the BmKMO gene was analyzed, and three guide RNAs (gRNAs) were prepared. Each synthesized gRNA was combined with Cas9 protein and then analyzed by T7 endonuclease I after introduction into the BM-N silkworm cell line. To edit the silkworm gene, K1P gRNA and Cas9 complexes were subsequently microinjected into the silkworm embryos; the hatching rate was 18% and the incidence of mutation was 60%. The gene mutation was verified in the heterozygous G0 generation, but no phenotypic change was observed. In homozygotes generated by self-crossing, a mutant phenotype was observed. These results suggest that silkworm molecular breeding using the CRISPR/Cas9 system is possible and could be an effective way of shortening the time required.

• Investigative Magnetic Resonance Imaging
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• v.2 no.1
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• pp.96-103
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• 1998

Purpose : To assess the clinical utility of turbo contrast-enhanced magnetic resonance angiography(CE MRA) in the evaluation of the aortic arch and its major branches and to compare the image quality of CE MRA among different coils used. Materials and Methods : Turbo three-phase dynamic CE MRA encompassing aortic arch and its major branches was prospectively performed after manual bolus IV injection of contrast material in 29 patients with suspected cerebrovascular diseases at 1.0T MR unit. the raw data were obtained with 3-D FISH sequence (TR 5.4ms, TE 2.3ms, flip angle 30, slab thickness 80nm, effective slice thickness 4.0mm, matrix size $100{\times}256$, FOV 280mm). Total data acquisition time was 4. to 60 seconds. We subjectively evaluated the imge quality with three-rating scheme : "good" for unequivocal normal finding, "fair" for relatively satisfactory quality to diagnose 'normal' despite intravascular low signal, and "poor" for equivocal diagnosis or non-visualization of the origin or segment of the vessels due to low signal or artifacts which needs catheter angiography. At the level of the carotid bifurcation, it was compared with conventional 2D-TOF MRA image. Overall image quality was also compared visually and quantitatively by measuring signal-to-noise ratios (SNRs) of the ascending aorta, the innominate artery and both common carotid arteries among the three different coils used(CP body array(n=12), CP neck array(n=9), and head-and-neck(n=8). Results : Demonstration of the aortic arch and its major branches was rated as "good" in 55% (16/29) and "fair" in 34%(10/29). At the level of the carotid bifurcation, image quality of turbo CE MRA was same as or better than conventional 2D-TOF MRA in 65% (17/26). Overall image quality and SNR were significantlygreater with CP body array coil than with CP neck array or head-and-neck coil. Conclusions : Turbo CE MRA can be used as a screening exam in the evaluation of the major branches of the aortic arch from their origin to the skull base. Overall imagequality appears to be better with CP body array coil than with CP neck array coil or head-and-neck coil.

• Korean Journal of Microbiology
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• v.44 no.3
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• pp.237-243
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• 2008

Chemical and microbial characteristics of bacterial populations were investigated in a quercus and pine humus forest soil. Soil pH was $5.3\pm0.4$ and $4.1\pm0.9$ from each sample of a quercus and pine humus forest soil; C/N ratio of humus forest soil was $17.84\pm4.6%$ and $21.76\pm8%$, respectively. Total organic acid was investigated as 69.57 mM/g dry soil and 53.72 mM/g dry soil in each humus forest soil. Glutamine, pyruvate, succinate, lactic acid and acetic acid of pine humus forest soil were $1.5\sim4.5$ times higher than those of quercus humus forest soil. As we evaluated phylogenetic characteristics of bacterial populations by 16S rRNA-ARDRA analysis with DNA extracted from each humus forest soil. Based on the 16S rRNA sequences, 44 clone from ARDRA groups of quercus humus forest soil were classified into 7 phyla: ${\alpha},{\beta},{\gamma},{\delta}$-Proteobacteria, Acidobacteria, Actinobacteria, and Firmicutes. Thirty-two clone from ARDRA groups of pine humus forest soil were classified into 8 phyla: ${\alpha},{\beta},{\gamma}$-Proteobacteria, Acidobacteria, Bacteroides, Verrucomicrobia, Planctomycetes, and Gemmatomonadetes. According to PCA (Principal Component Analysis) based on 16S rRNA base sequence, there were three main groups of bacteria. All clone of Cluster I were originated from quercus humus forest soil, while 67% clone of Cluster II and 63% clone of Clusters III were separated from pine humus forest soil.

• Journal of Chest Surgery
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• v.33 no.3
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• pp.221-229
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• 2000

Background: Thromboxane A2 and endothelin-1 are the potent vasoconstrictors affecting pulmonary pathophysiology in response to whole body inflammatin following CPB. Aprotinin, as an antiiflammatory agent, may decrease the release of such vasoactive substance from pulmonary tissues, preventing pulmonary hypertension after cardiopulmonary bypass. Material and Method: Ten mongrel dogs(Bwt. ac. 20kg) were subjected to cardioupulmonary bypass for 2 hours and postbypass pulmonary vascular resistance(0, 1, 2, 3 hours) were compared with prebypass level. The dogs were divided into 2 groups; control group(n-5) and aprotinin group(n=5). In the aprotinin group, aprotinin was administered as follows; 50,000 KIU/kg mixed in pump priming solution, 50,000 KIU/kg prebypass intravenous infusion over 30 minutes, 10,000 KIU/kg/hour postbypass continuous infusion. Prebypass and postbypass 0, 1, 2, 3 hour pulmonary vascular resistance were measured. At prebypass and postbypass 0, 90, 180 minutes, blood samples were obtained from pulmonary arterial and left atrial catherers for the assay of plasma thromboxane B2 a stable metabolite of thromboxane A2, and endothelin-1 concentrations. Result: The ratios of pustbypass over prebypass pulmonary vascular at postbypass 0, 1, 2, 3 hours were 1.28$\pm$0.20, 1.82$\pm$0.23, 1.90$\pm$0.19, 2.14$\pm$0.18 in control group, 1.58$\pm$0.18, 1.73$\pm$0.01, 1.66$\pm$0.10, 1.50$\pm$0.08 in aprotinin group ; the ratios gradually increased in control group while decreased or fluctuated after postbypass 1 hour in aprotinin group. There was statistically significant difference between control group and aprotinin group at postbypass 3 hours(P=0.014). Pulmonary arterial plasma concentration of thromboxane B2(pg/ml) at prebypass, postbypass 0, 90, 180 minutes were 346.4$\pm$61.9, 529.3$\pm$197.6, 578.3$\pm$255.8, 493.3$\pm$171.3 in control group, 323.8$\pm$118.0, 422.6$\pm$75.6, 412.3$\pm$59.9, 394.5$\pm$154.0 in aprotinin group. Left atrial concentrations were 339.3$\pm$89.2, 667.0$\pm$65.7, 731.2$\pm$192.7, 607.5$\pm$165.9 in control group, 330.0$\pm$111.2, 468.4$\pm$190.3, 425.4$\pm$193.6, 4.7.3$\pm$142.8 in aprotinin group. These results showed decrement of pulmonary thromboxane A2 generation in aprotinin group. Pulmonary arterial concentrations of endothelin-1(fmol/ml) at the same time sequence were 7.84$\pm$0.31, 13.2$\pm$0.51, 15.0$\pm$1.22, 16.3$\pm$1.73 in control group, 7.76$\pm$0.12, 15.3$\pm$0.71, 22.6$\pm$6.62, 14.9$\pm$1.11 in aprotinin group. Left atrial concentrations were 7.61$\pm$17.2, 57.1$\pm$28.4, 18.9$\pm$18.2, 31.5$\pm$20.5 in control group, 5.61$\pm$7.61, 37.0$\pm$26.2, 28.6$\pm$21.7, 37.8$\pm$30.6 in aprotinin group. These results showed that aprotinin had no effect on plasma endothelin-1 concentration after cardiopulmonary bypass. Conclusion: Administration of aprotinin during cardiopulmonary bypass could attenuate the increase in pulmonary vascular resistance after bypass. Inhibition of pulmonary thromboxane A2 generation was thought to be one of the mechanism of this effect. Aprotinin had no effect on postbypass endothelin-1 concentration.

• Journal of radiological science and technology
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• v.34 no.3
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• pp.239-244
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• 2011

In this study, we evaluated diffusion weighted imaging (DWI) to investigate whether the DWI parameters can predict characteristic parameters on pathologic specimens of tumor or not. CFPAC-1 was injected subcutaneously on the back flank of athymic nude mice (n=13) then two tumors were initiated on each mouse (2${\times}$13=26 tumors). The mice were sacrificed to make specimen immediately after initial MR imaging then were compared with the MR image. A dedicated high-field (7T) small-animal MR scanner was used for image acquisitions. A T1 and T2 weighted axial image using RARE technique was acquired to measure the T2 values and tumor size. DWI MR was performed for calculating ADC values. To evaluate tumor cellularity and determine the levels of MVD, tumor cells were excised and processed for H-E staining and immunostaining using CD31. T2 values and ADC values were computed and analyzed for each half of the tumors and compared to the correlated specimens slide. Median ADC within each half of mass was compared to the cellularity and MVD in the correlated area of pathologic slide. The mean of ADC value is $0.7327{\times}10^{-3}$ $mm^2/s$ and standard deviation is $0.1075{\times}10^{-3}$ $mm^2/s$. There is a linear relationship between ADC value and tumor necrosis (R2=0.697, p< 0.001). DW image parameters including the ADC values can be utilized as surrogate markers to assess intratumoral neoangiogenesis and change of the internal structure of tumor cells.

• Journal of Life Science
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• v.28 no.12
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• pp.1416-1423
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• 2018

With strong biotin binding affinity ($K_D=10^{-14}M$), the tetrameric feature of streptavidin could be used to increase the antigen binding activity of a camel heavy chain (VHH) antibody through their fusion, here stained with biotinylated horseradish peroxidase and subsequent immunoassays ELISA and Western blot analysis. For this application, we cloned the streptavidin gene amplified from the Streptomyces avidinii chromosome by PCR, and this was fused to the gene of the 8B9 VHH antibody which is specific to green fluorescent protein (GFP) antigens. To express a soluble fusion protein in Escherichia coli, we used the pUC119 plasmid-based expression system which uses the lacZ promoter for induction by IPTG, the pelB leader sequence at the N-terminus for secretion into the periplasmic space, and six polyhistidine tags at the C-terminus for purification of the expressed proteins using an $Ni^+$-NTA-agarose column. Although streptavidin is toxic to E. coli because of its strong biotin binding property, this soluble fusion protein was expressed successfully. In SDS-PAGE, the size of the purified fusion protein was 122.4 kDa in its native condition and 30.6 kDa once denatured by boiling, suggesting the tetramerization of the monomeric subunit by non-covalent association through the streptavidin moiety fusing to the 8B9 VHH antibody. In addition, this fusion protein showed biotin binding activity similar to streptavidin as well as GFP antigen binding activity through both ELISA and Western blot analysis. In conclusion, the protein resulting from the fusion of an 8B9 VHH antibody with streptavidin was successfully expressed and purified as a soluble tetramer in E. coli; it showed both biotin and GFP antigen binding activity suggesting the possible production of a tetrameric and bifunctional VHH antibody.

• Journal of Life Science
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• v.25 no.6
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• pp.680-685
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• 2015

Previously, cellulase and xylanase producing microorganism, Bacillus subtilis NC1, was isolated from soil. Based on the 16S rRNA gene sequence and API 50 CHL test the strain was identified as Bacillus subtilis, and named as B. subtilis NC1. We cloned and sequenced the genes for cellulase and xylanase. Plus, the deduced amino acid sequences from the genes of cellulase and xylanase were determined and were also identified as glycosyl hydrolases family (GH) 5 and 30, respectively. In this study to optimize the medium parameters for cellulase production by B. subtilis NC1 the RSM (response surface methodology) based on CCD (central composite design) model was performed. Three factors, tryptone, yeast extract, and NaCl, for N or C source were investigated. The cellulase activity was measured with a carboxylmethyl cellulose (CMC) plate and the 3,5-dinitrosalicylic acid (DNS) methods. The coefficient of determination (R²) for the model was 0.960, and the probability value (p=0.0001) of the regression model was highly significant. Based on the RSM, the optimum conditions for cellulase production by B. subtilis NC1 were predicted to be tryptone of 2.5%, yeast extract of 0.5%, and NaCl of 1.0%. Through the model verification, cellulase activity of Bacillus subtilis NC1 increased from 0.5 to 0.62 U/ml (24%) compared to the original medium.