Tuberculosis, caused by Mycobacterium tuberculosis, continues to be one of the leading infectious diseases to humans. It is urgent to discover novel drug targets for the development of antitubercular agents. The 2-C-methyl-Derythritol-4-phosphate (MEP) pathway for isoprenoid biosynthesis has been considered as an attractive target for the discovery of novel antibiotics for its essentiality in bacteria and absence in mammals. MEP cytidyltransferase (IspD), the third-step enzyme of the pathway, catalyzes MEP and CTP to form 4-diphosphocytidyl-2-C-methylerythritol (CDP-ME) and PPi. In the work, ispD gene from M. tuberculosis H37Rv (MtIspD) was cloned and expressed. With N-terminal fusion of a histidine-tagged sequence, MtIspD could be purified to homogeneity by one-step nickel affinity chromatography. MtIspD exists as a homodimer with an apparent molecular mass of 52 kDa. Enzyme property analysis revealed that MtIspD has high specificity for pyrimidine bases and narrow divalent cation requirements, with maximal activity found in the presence of CTP and $Mg^{2+}$. The turnover number of MtIspD is $3.4 s^{-1}$. The Km for MEP and CTP are 43 and $92{\mu}M$, respectively. Furthermore, MtIspD shows thermal instable above $50^{\circ}C$. Circular dichroism spectra revealed that the alteration of tertiary conformation is closely related with sharp loss of enzyme activity at higher temperature. This study is expected to help better understand the features of IspD and provide useful information for the development of novel antibiotics to treat M. tuberculosis.
This study tried to evaluate the difference of stress degree by Sasang constitution and lifestyle for workplace labors, and suggested the preventive oriental medicine approaches in occupational health area. 479 data for research were collected through the process of oriental health examination with questionnaire in workplace, and collected data were analyzed with frequency, crosstab, and Two-Way ANOVA. The results were as follows: 1) All subjects, total 479 were men, and constitutional distributions were 39.2% of Taeumin, 32.4% of Soumin, and 28.4% of Soyangin. The distributions of stress score were 13.2% of normal group, 13.4% of high risk group, and 73.5% of potential risk group. 2) There were no statistically significant differences for stress by constitution(p=0.085). By the results of optimal scaling and homogeneity analysis, Soyangin was close to potential risk group compared to other constitution, and Soumin and Taeumin were close to high risk and normal group, respectively. 3) For the difference of stress score by Sasang constitution and lifestyle, Soumin who exercised nearly everyday had the highest stress score with interaction in Factor 4 area(p<0.05), and there was no interaction in other area. 4) For the difference of PWI score by lifestyle, there were statistically significant differences for drinking(p<0.01), smoking(p<0.01), and obesity index(p<0.01)
In Korea, all domestic made test systems for detecting antibodies in HIV-1 contain the antigens from human immunodeficiency type 1 (HIV-1) subtype B. However, because HIV-1 subtype O is significantly different in amino acid sequences from all other subtypes of HIV-1, there has been a need for developing a test for detecting antibodies in subtype O. For this purpose, the entire nucleotide sequence corresponding to the extracellular domain of the transmembrane glycoprotein of HIV-1 subtype O was synthesized with consideration of Escherichia coli condon usage. Various regions of the extracellular domain were cloned into E. coli expression vectors and tested for levels of protein production. The nucleotide sequence, named ECTM, that can encode a 129 amino acid-long peptide, was found to be expressed at a high level in E. coli. The protein of approximately 17 kDa specifically reacted with sera from individuals infected with HIV-1 subtype O. The ECTM protein was purified to near homogeneity by the CM-T gel chromatography, using concentrated, denatured inclusion bodies. In Western blot analysis, the purified viral antigen reacted with sera from individuals infected with subtype O more efficiently than subtype B. The enzyme linked immunoabsorbent assay (ELISA) system was developed using the subtype O viral protein and compared with the commercially available kit lacking the antigens from subtype O. The ELISA kit containing the subtype O antigen ECTM alone efficiently reacted with sera from individuals infected with subtype O. The subtype O antigen-containing kit produced a positive absorbence even when sera were diluted 512-fold, suggesting a high sensitivity. The commercially available kit also reacted with subtype O sera, but produced a negative result at a dilution of 8-fold. Our results suggest that the currently available kit may not be able to efficiently detect subtype O sera and that the viral protein developed in this study may be added to the current system to maximize the detection of sera from individuals infected with subtype O.
Kim, Do-Hyoung;Park, Jeong-Uck;Seo, Min-Jeong;Kim, Min-Jeong;Lee, Hye-Hyeon;Choi, Yung-Hyun;Joo, Woo-Hong;Jeong, Yong-Kee
Journal of Life Science
/
v.20
no.2
/
pp.183-189
/
2010
A fibrinolytic enzyme producing Bacillus sp. J-19 was isolated from the popular Korean seasoning, pickled anchovy. The fibrinolytic enzyme was purified to homogeneity by chromatographic methods including ethanol precipitation and gel-filtration using Sephadex G-50. Compared to the crude enzyme extract, the specific activity of the enzyme increased 1021-fold with a recovery of 23%. The purified enzyme was estimated to be approximately 45 kDa by SDS-PAGE. Especially, the amidolytic activity in the presence of the synthetic substrate for serine protease (H-D-Ile-Pro-Arg-pNA, S-2288) represented approximately 17 U/mg. In addition, more than the 60% activity of the 45 kDa fibrinolytic activity was maintained in the presence of up to 30% (w/v) sodium chloride. These findings could provide a unique fibrinolytic enzyme, leading to a potential thrombolytic agent.
Hong, Jin Ho;Lee, Ha Young;Kang, Young Hye;Lim, Myung Kwan;Kim, Yeo Ju;Cho, Soon Gu;Kim, Mi Young
Investigative Magnetic Resonance Imaging
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v.20
no.1
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pp.44-52
/
2016
Purpose: To quantitatively and qualitatively compare fat-suppressed MRI quality using iterative decomposition of water and fat with echo asymmetry and least-squares estimation (IDEAL) with that using frequency selective fat-suppression (FSFS) T2- and postcontrast T1-weighted fast spin-echo images of the head and neck at 3T. Materials and Methods: The study was approved by our Institutional Review Board. Prospective MR image analysis was performed in 36 individuals at a single-center. Axial fat suppressed T2- and postcontrast T1-weighted images with IDEAL and FSFS were compared. Visual assessment was performed by two independent readers with respect to; 1) metallic artifacts around oral cavity, 2) susceptibility artifacts around upper airway, paranasal sinus, and head-neck junction, 3) homogeneity of fat suppression, 4) image sharpness, 5) tissue contrast of pathologies and lymph nodes. The signal-to-noise ratios (SNR) for each image sequence were assessed. Results: Both IDEAL fat suppressed T2- and T1-weighted images significantly reduced artifacts around airway, paranasal sinus, and head-neck junction, and significantly improved homogeneous fat suppression in compared to those using FSFS (P < 0.05 for all). IDEAL significantly decreased artifacts around oral cavity on T2-weighted images (P < 0.05, respectively) and improved sharpness, lesion-to-tissue, and lymph node-to-tissue contrast on T1-weighted images (P < 0.05 for all). The mean SNRs were significantly improved on both T1- and T2-weighted IDEAL images (P < 0.05 for all). Conclusion: IDEAL technique improves image quality in the head and neck by reducing artifacts with homogeneous fat suppression, while maintaining a high SNR.
Journal of the Korea Society of Computer and Information
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v.16
no.12
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pp.297-304
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2011
This study was to analyze the learning effectiveness of multi-media based class by comparing with traditional classroom method. The "Distribution Working Subject" course that is one of the required courses of Vocational high school was selected and its contents were digitalized on MS Powerpoint for multi-media based class. The thirty students were sampled for each experimental and control groups. The homogeneity and learning achievement of sample groups were tested for experiment. Same teacher took the classes of two groups and delivered same contents of course. Only difference between two groups was the delivery method, one is traditional classroom teaching method and the other was the multi-media based class. The learning achievements and satisfaction of sample were post-tested in order to analyze the learning effectiveness by comparing two teaching methods. The results showed that there was a significant difference between experimental and control group in learning achievement after ANCOVA controlled pre-test as covariance(F=5.08, p<.05). It means that the learning achievement of multi-media based class was higher than that of traditional classroom group. The results also showed that a significant difference in students' satisfaction between two groups (t=5.57, p<.001). This study concluded that using multi-media in class could produce more learning achievements and satisfaction of students than traditional classroom method.
The Tongcheng pig breed is a famous Chinese indigenous breed. The Ministry of Agriculture of China has filed it as 1 of 19 national key conservation breeds selected from more than 100 Chinese indigenous pig breeds in 2000. In order to improve the reproductive performance, it has been intensively selected to increase the litter size for about 10 years. The population randomly sampled from conservation nucleus of eight families in the Tongcheng pigs was genotyped for identification of their estrogen receptor locus polymorphisms with the PCR-RFLPs method. Only AB heterozygotes and BB homozygotes were detected, and $X^2$ test demonstrated that the locus was in disequilibrium at a significant level (p<0.05). In the present paper, the litter sizes in different parities were regarded as different traits. Holistic status of other unspecific and unidentified genes was estimated by using the statistical methods. Coefficients of kurtosis and skewness showed that the litter size still presented segregating characteristic in the 2nd, 5th, 7th, 8th and 9th parities. Analysis of homogeneity of variance between families confirmed the results for the 5th, 7th and 8th parities. The heritability of litter size for the 1st to 10th parities was estimated with paternal half-sib model and individual estimated breeding values (EBVs) were evaluated by a single trait animal model as well. We found that the averages of EBVs for litter size in each parity did not differ significantly between genotypes, despite the significant difference for original phenotype records in the 3rd, 4th and 5th parities (p<0.05 or p<0.01). The results may be explained by the deduction that the polymorphisms of ESR locus are no longer the important genetic base of litter size variation when the frequency of allele B accumulated in the experience of selection procedure, and further conferring that there exist special genes associated with litter size in the recent Tongcheng pigs population can be made.
Yang, Jeong Sun;Lee, Mi Young;Park, In Jeong;Kang, Seong-Kyu
Analytical Science and Technology
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v.13
no.2
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pp.258-267
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2000
The result of five year's experience on Inter-laboratory Comparison for Analyses of Heavy Metals and Organic Solvent Metabolites in Biological Samples was described. Since 1995, around a hundred laboratories in the Occupational Health have participated this program twice per year by the Industrial Safety and Health Law. Four metals in blood and five organic solvent metabolites in urine were examined. Reference samples were made by spiking standard materials to human blood or urine pools treated previously to give homogeneity and stability for a specific time periods. Some reference samples for organic solvent metabolites were made from workers' urine who were exposed to the organic solvents. Some items such as Lead in blood and Hippuric acid in urine showed good accordance between participants while the other items such as Mercury and N-methylformamide in urine showed poor proficient rate. The results were published in the internet or newspaper to help the consumer of the laboratory's service to get the information on them and to make competition between them. The inter-laboratory's comparison program have done greate role to improve the ability of analysis and reliability of analytical data produced from each laboratory.
Exo-xylanase encoded by the xylA gene of Bacillus stearothermoPhillus was produced from Escherichia coli ]M109 carrying a recombinant plasmid pMGL Synthesis of the enzyme was observed to be cell-associated, and about 94% of the enzyme synthesized was located in the cytoplasmic region. The maximum production was attained when the E. coli strain was grown at $37^{\circ}C$ for 8 hours on the medium containing 0.5% fructose, 1.0% tryptone, 1.0% sodium chloride, and 0.5% yeast extract. The exo-xylanase was purified to homogeneity using a combination of salting out with ammonium sulfate, DEAE-Sepharose CL-6B ion exchange chromatography, Sephadex G-IOO gel filtration, and Sephadex G-150 gel filtration. The' purified enzyme was most active at pH 6.0 and $45^{\circ}C$. $Ca^{2+}$ and $Co^{2+}$ activated the exo-xylanase activity by about 20% while $Ag^{2+}$, $Fe^{2+}$, $Mg^{2+}$ and $Zn^{2+}$ inhibited the enzyme activity by up to 60%. The $K_m$, value on p-nitrophenyl-$\beta$-D-xylanopyranoside was 2.75 mM. The enzyme had a pI value of 4.7. The estimated molecular weight of the native protein was 200,000 daL SDS-polyacrylamide gel electrophoresis analysis suggested that the native enzyme was a trimer composed of three identical 66,000 da!. polypeptides. The purified enzyme efficiently converted all the xylo-oligosaccharides tested to xylose. It was also confirmed that the enzyme split xylans in an exo-manner even though the degree of hydrolysis was fairly low. The xylanolytic enzyme was, therefore, classified to be one of the few bacterial exo-xylanases lacking transferase activity.
Purpose: The purpose of this study has attempted to evaluate and compare the image evaluation and exposure dose by respectively applying Filtered Back Projection(FBP), the existing test method, and Adaptive Statistical Iterative Reconstruction(ASIR) with different values of tube voltage during the Low Dose Computed Tomography(LDCT). Materials and Methods: With the image reconstruction method as basis, Chest Phantom was utilized with the FBP and ASIR set at 10%, 20% respectively, and the change of Tube Voltage (100kVp, 120kVp). For image evaluation, Back ground noise, Signal to Noise ratio(SNR) and Contrast to Noise ratio(CNR) were measured, and, for dose evaluation, CTDIvol and DLP were measured respectively. The statistical analysis was tested with SPSS(ver. 22.0), followed by ANOVA Test conducted after normality test and homogeneity test. (p<0.05). Results: In terms of image evaluation, there was no outstanding difference in Ascending Aorta(AA) SNR and Infraspinatus Muscle(IM) SNR with the different values of ASIR application(p<0.05), but a significant difference with the different amount of tube voltage(p>0.05). Also, there wasn't noticeable change in CNR with ASIR and different amount of Tube Voltage (p<0.05). However, in terms of dose evaluation, CTDIvol and DLP showed contrasting results(p<0.05). In terms of CTDIvol, the measured values with the same tube voltage of 120kVp were 2.6mGy with No-ASIR and 2.17mGy with 20%-ASIR respectively, decreased by 0.43mGy, and the values with 100kVp were 1.61mGy with No-ASIR and 1.34mGy with 20%-ASIR, decreased by 0.27mGy. In terms of DLP, the measured values with 120kVp were $103.21mGy{\cdot}cm$ with No-ASIR and $85.94mGy{\cdot}cm$ with 20%-ASIR, decreased by $17.27mGy{\cdot}cm$(about 16.7%), and the values with 100kVp were $63.84mGy{\cdot}cm$ with No-ASIR and $53.25mGy{\cdot}cm$ with 20%-ASIR, a decrease by $10.62mGy{\cdot}cm$(about 16.7%). Conclusion: At lower tube voltage, the rate of dose significantly decreased, but the negative effects on image evaluation was shown due to the increase of noise. For the future, through the result of the experiment, it is considered that the method above would be recommended for follow-up patients or those who get health checkup as long as there is no interference on the process of diagnosis due to the characteristics of Low Dose examination.
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