• Journal of Life Science
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• v.28 no.11
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• pp.1301-1315
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• 2018

We purified an antimicrobial peptide from the acidified hemocyte extract of Mytilus coruscus by $C_{18}$ reversed-phase high-performance liquid chromatography (RP-HPLC). The peptide was 4041.866 Da based on matrix-assisted laser desorption ionization time-of-flight mass spectrophotometer (MALDI-TOF/MS) and the 25 amino acids of the N-terminus sequence were identified. Comparison of this sequence of the purified peptide with the N-terminus sequences of other antimicrobial peptides revealed 100% identity with the mytilin B precursor of Mytilus coruscus. We also identified a 312 bp open-reading frame (ORF) encoding 103 amino acids based on the obtained amino acid residues. The nucleotide sequence of this ORF and the amino acid sequence also revealed 100% identity with the mytilin B precursor of Mytilus coruscus. We synthesized two antimicrobial peptides with an alanine residue in the C-terminus, and designated them mytilin B1 and B2. These two antimicrobial peptides showed antimicrobial activity against gram-positive bacteria, including Bacillus cereus and Streptococcus parauberis (minimal effective concentration, MECs $41.6-89.7{\mu}g/ml$), gram-negative bacteria, including Enterobacter cloacae, Escherichia coli, Klebsiella pneumoniae, Proteus mirabilis, Providencia stuartii, Pseudomonas aeruginosa, and Vibrio ichthyoenteri (MECs $7.4-39.5{\mu}g/ml$), and the fungus Candida albicans (MECs $26.0-31.8{\mu}g/ml$). This antimicrobial activity was stable under heat and salt conditions. Furthermore, the peptides did not exhibit significant hemolytic activity or cytotoxic effects. These results suggest that mytilin B could be applied as alternative antibiotic agent, and they add to the understanding of the innate immunity of hard-shelled mussels.

• Analytical Science and Technology
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• v.20 no.4
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• pp.308-314
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• 2007

The analytical method of HPLC with the radial-flow electrochemical cell (RFEC) has been developed to determine doxorubicin, epirubicin, nogalamycin, daunorubicin and idarubicin simultaneously by employing a reversed-phase chromatography. Anthracyclines were detected at -0.74 V vs. a Ag/AgCl (0.01 M NaCl) reference electrode, a potential of diffusion current plateau in the mobile phase. At a $V_f$ of 1.0 mL/min doxorubicin, epirubicin, daunorubicin and idarubicin appeared at a retention time ($t_r$) of 6.4 min, 7.4 min, 12.7 min and 18.4 min, respectively, while at a $V_f$ of 0.6 mL/min, doxorubicin, epirubicin, nogalamycin, daunorubicin and idarubicin appeared at a $t_r$ of 9.9 min, 11.5 min, 13.5 min, 19.6 min and 28.7 min, respectively. The linearity between each anthracycline injected ($2.40{\times}10^{-7}M{\sim}1.42{\times}10^{-5}M$) and peak area (charge) was excellent with the square of the correlation coefficient ($R^2$) higher than 0.999. The detection limits were $1.0{\times}10^{-8}M{\sim}1.5{\times}10^{-7}M$ for the five anthracyclines. Within-day precision for the five anthracyclines were in reasonable relative standard deviations less than 3 % ($1.00{\times}10^{-6}M{\sim}1.42{\times}10^{-5}M$) except the lower concentrations less than $0.7{\mu}M$. Solid phase extractions of $1.00{\times}10^{-5}M$ epirubicin, $0.48{\times}10^{-5}M$ nogalamycin and $1.52{\times}10^{-5}M$ daunorubicin from human serum with a $C_{18}$ cartridge resulted in 97 %, 100 % and 90 % of recoveries, respectively.

• Nuclear Medicine and Molecular Imaging
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• v.40 no.4
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• pp.218-227
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• 2006

Purpose: The HSV1-tk reporter gene system is the most widely used system because of its advantage that direct monitoring is possible without the introduction of a separate reporter gene in case of HSV1-tk suicide gene therapy. In this study, we investigate the usefulness of the reporter probe (substrate), $9-(4-[^{18}F]Fluoro-3-hydroxymethylbutyl)$guanine ($[^{18}F]FHBG$) for non-invasive reporter gene imaging using PET in HSV1-tk expressing hepatoma model. Materials and Methods: Radiolabeled FHBG was prepared in 8 steps from a commercially available triester. The labeling reaction was carried out by NCA nucleophilic substitution with $K[^{18}F]/K2.2.2.$ in acetonitrile using N2-monomethoxytrityl-9-14-(tosyl)-3-monomethoxytritylmethylbutyl]guanine as a precursor, followed by deprotection with 1 N HCl. Preliminary biological properties of the probe were evaluated with MCA cells and MCA-tk cells transduced with HSV1-tk reporter gene. In vitro uptake and release-out studies of $[^{18}F]FHBG$ were performed, and was analyzed correlation between $[^{18}F]FHBG$ uptake ratio according to increasing numeric count of MCA-tk cells and degree of gene expression. MicroPET scan image was obtained with MCA and MCA-tk tumor bearing Balb/c-nude mouse model. Results: $[^{18}F]FHBG$ was purified by reverse phase semi-HPLC system and collected at around 16-18 min. Radiothemical yield was about 20-25%) (corrected for decay), radiochemical purity was >95% and specific activity was around >55.5 $GBq/{\mu}\;mol$. Specific accumulation of $[^{18}F]FHBG$ was observed in HSV1-tk gene transduced MCA-tk cells but not in MCA cells, and consecutive 1 hour release-out results showed more than 86% of uptaked $[^{18}F]FHBG$ was retained inside of cells. The uptake of $[^{18}F]FHBG$ was showed a highly significant linear correlation ($R^2=0.995$) with increasing percentage of MCA-tk numeric cell count. In microPET scan images, remarkable difference of accumulation was observed for the two type of tumors. Conclusion: $[^{18}F]FHBG$ appears to be a useful as non-invasive PET imaging substrate in HSV1-tk expressing hepatoma model.

• Journal of Korean Society of Environmental Engineers
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• v.31 no.2
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• pp.79-89
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• 2009

Naphthalene is a volatile, hydrophobic, and possibly carcinogenic compound that is known to have a severe detrimental effect to aquatic ecosystem. Our research examined the effects of various operating conditions (temperature, pH, initial concentration, and frequency and type of ultrasound) on the sonochemical degradation of naphthalene and OH radical production. The MDL (Method detection limit) determined by LC/FLD (1200 series, Agilient) using C-18 reversed column is measured up to 0.01 ppm. Naphthalene vapor produced from ultrasound irradiation was detected under 0.05 ppm. Comparison of naphthalene sonodegradion efficiency tested under open and closed reactor cover fell within less than 1% of difference. Increasing the reaction temperature from $15^{\circ}C$ to $40^{\circ}C$ resulted in reduction of naphthalene degradation efficiency ($15^{\circ}C$: 95% ${\rightarrow}$ $40^{\circ}C$: 85%), and altering pH from 12 to 3 increased the effect (pH 12: 84% ${\rightarrow}$pH 3: 95.6%). Pseudo first-order constants ($k_1$) of sonodegradation of naphthalene decreased as initial concentration of naphthalene increased (2.5 ppm: $27.3{\times}10^{-3}\;min^{-3}\;{\rightarrow}$ 10 ppm : $19.3{\times}10^{-3}\;min^{-3}$). Degradation efficiency of 2.5 ppm of naphthalene subjected to 28 kHz of ultrasonic irradiation was found to be 1.46 times as much as when exposed under 132 kHz (132 kHz: 56%, 28 kHz: 82.7%). Additionally, its $k_1$ constant was increased by 2.3 times (132 kHz: $2.4{\times}10^{-3}\;min^{-1}$, 28 kHz: $5.0{\times}10^{-3}\;min^{-1}$). $H_2O_2$ concentration measured 10 minutes after the exposure to 132 kHz of ultrasound, when compared with the measurement under frequency of 28 kHz, was 7.2 times as much. The concentration measured after 90 minutes, however, showed the difference of only 10%. (concentration of $H_2O_2$ under 28 kHz being 1.1 times greater than that under 132 kHz.) The $H_2O_2$ concentration resulting from 2.5 ppm naphthalene after 90 minutes of sonication at 24 kHz and 132 kHz were lower by 0.05 and 0.1 ppm, respectively, than the concentration measured from the irradiated M.Q. water (no naphthalene added.) Degradation efficiency of horn type (24 kHz) and bath type (28 kHz) ultrasound was found to be 87% and 82.7%, respectively, and $k_1$ was calculated into $22.8{\times}10^{-3}\;min^{-1}$ and $18.7{\times}10^{-3}\;min^{-1}$ respectively. Using the multi- frequency and mixed type of ultrasound system (28 kHz bath type + 24 kHz horn type) simultaneously resulted in combined efficiency of 88.1%, while $H_2O_2$ concentration increased 3.5 times (28 kHz + 24 kHz: 2.37 ppm, 24 kHz: 0.7 ppm.) Therefore, the multi-frequency and mixed type of ultrasound system procedure might be most effectively used for removing the substances that are easily oxidized by the OH radical.

• Journal of Preventive Medicine and Public Health
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• v.22 no.4 s.28
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• pp.506-517
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• 1989

Two hundred and fifty teachers of special school (for the disabled) and 414 elementary school teachers were selected for the targets in order to compare their degrees of fatigue symptoms and to find what kind of ralationship is between subjective symptoms of fatigue and pH is the saliva. It was 30 minutes before their closing hours on April 21th, 1989 that their physical, mental and neuro-sensory symptoms and salivary pH were examined. The test results are summarized as fallows : It is observed that an interrelation between subjective fatigue and pH in their saliva shows a significant relationship between physical and neuro-sensory symptoms in a sense of statistics. The rate of subjective fatigue complained by the special teachers is higher than that by the elementary teachers. In the case of salivary pH, the special teachers' is as a whole lower than the elementary teachers'. The complain rates in each item, checked, of special teachers are generally higher than those of the elementary teachers. It is in the mental symptom related item that there are many sub-items which show significant difference. According to the average of salivary pH based on the degrees of complained symptoms shown in the pH related items, the salivary pH of the group with complained symptoms is lower than that of the group without complained symptoms. In the rate of complaints, by sex, both sexes of the special teachers show high ones, but salivary pH is low. The complain rate of mental symptoms shown by female group from the special teacher Is significantly higher(p<0.05). By age, the group in their thirties from the special teachers show the higher complain rate of mental symptoms (29.3%) and the lower salivary pH (p<0.05) than that (15.1%) of the elementary teachers belonging to the same age catagory. However, the special teachers in their forties show the lower complain rate of physical symptoms that of the elementary teachers (p<0.05). From the viewpoint of their working years, the special teachers below 14 years and elementary teachers above 15 years in their career show high complain rates. Among those who belong to the catagory of 10-14 working years, the special teachers show the higher complain rate of mental symptoms than that of their counterparts. In the case of the salivary pH, the special teachers of all working-year catagories show the higher pH than that of the elementary teachers. But there is not significantly difference. From the viewpoint of sleeping hours in the previous night of the questionaire surveyed, among those who slept for over 7 hours, the special teachers show the higher complain rate of mental symptoms with a significant difference, but the lower salivary pH than that of their counterparts. From the viewpoint of their marital status, existence of disease history, the special teachers show the higher complain rate of subjective fatigue, but the lower salivary pH than that of the elementary teachers respectively. According to the above results, the special teachers generally show the higher complain rate of subjective fatigue, the lower salivary pH, and the higher complain rate of mental symptoms. To prevent the possible accumulation of mental fatigue of the special teachers, ways and means to make use of leisure time, recreational facilities are necessarily provided. Since the degree of fatigue and salivary pH have a correlation to some extent, it is necessary that further continuous studies on the correlation between the degrees of fatigue and salivary pH should be pursued.