Journal of the Korea Academia-Industrial cooperation Society
/
v.13
no.11
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pp.5350-5355
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2012
Human tyrosinase (hTyr) catalyzes the first and rate limiting step in the biosynthesis of a skin color determinant, melanin. Although a number of cosmetic companies have tried to develop hTyr inhibitors for several decades, absence of 3D structure of hTyr make it impossible to design or screen inhibitors by structure-based approach. Therefore, we built a 3D structure by comparative modeling technique based on the crystal structure of tyrosinase from Bacillus megaterium to provide structural information and to search new hit compounds from database. Our model revealed that two copper atoms of active site located deep inside and were coordinated with six strictly conserved histidine residues coming from four-helix-bundle. Substrate binding site had narrow funnel like shape and its entrance was wide and exposed to solvent. In addition, hTyr-tyrosine and hTyr-kojic acid, a well-known inhibitor, complexes were modeled with the guide of solvent accessible surface generated by in-house software. Our model demonstrated that only phenol group or its analogs could fill the binding site near the nuclear copper center, because inside of binding site had narrow shape relatively. In conclusion, the results of this study may provide helpful information for designing and screening new anti-melanogenic agents.
Song-Ee Baek;Asad Ul-Haq;Dae Hee Kim;Hyoung Wook Choi;Myeong-Jin Kim;Hye Jin Choi;Honsoul Kim
Korean Journal of Radiology
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v.21
no.6
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pp.726-735
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2020
Objective: Recent innovations in biology are boosting gene and cell therapy, but monitoring the response to these treatments is difficult. The purpose of this study was to find an MRI-reporter gene that can be used to monitor gene or cell therapy and that can be delivered without a viral vector, as viral vector delivery methods can result in long-term complications. Materials and Methods: CMV promoter-human organic anion transporting polypeptide 1B3 (CMV-hOATP1B3) cDNA or CMV-blank DNA (control) was transfected into HEK293 cells using Lipofectamine. OATP1B3 expression was confirmed by western blotting and confocal microscopy. In vitro cell phantoms were made using transfected HEK293 cells cultured in various concentrations of gadoxetic acid for 24 hours, and images of the phantoms were made with a 9.4T micro-MRI. In vivo xenograft tumors were made by implanting HEK293 cells transfected with CMV-hOATP1B3 (n = 4) or CMV-blank (n = 4) in 8-week-old male nude mice, and MRI was performed before and after intravenous injection of gadoxetic acid (1.2 µL/g). Results: Western blot and confocal microscopy after immunofluorescence staining revealed that only CMV-hOATP1B3-transfected HEK293 cells produced abundant OATP1B3, which localized at the cell membrane. OATP1B3 expression levels remained high through the 25th subculture cycle, but decreased substantially by the 50th subculture cycle. MRI of cell phantoms showed that only the CMV-hOATP1B3-transfected cells produced a significant contrast enhancement effect. In vivo MRI of xenograft tumors revealed that only CMV-hOATP1B3-transfected HEK293 tumors demonstrated a T1 contrast effect, which lasted for at least 5 hours. Conclusion: The human endogenous OATP1B3 gene can be non-virally delivered into cells to induce transient OATP1B3 expression, leading to gadoxetic acid-mediated enhancement on MRI. These results indicate that hOATP1B3 can serve as an MRI-reporter gene while minimizing the risk of long-term complications.
The effects of Backryeoncho (Opuntia ficus-indica var. saboten) powder addition (0, 0.4, 0.8, and 1.2%) on the pH, titratable acidity, lactic acid contents, microbial contents, texture, and sensory evaluation of cabbage Kimchi, stored at $10^{\circ}C$ for 36 days were studied to establish an optimum recipe. Upon addition of Backryeoncho powder, pH was higher than that of Kimchi without powder after 36 days of fermentation. The titratable acidity was increased by the addition of Backryeoncho powder, and 0.4% powder added Kimchi showed the slowest changing level. During the fermentation period, the degree of salinity decreased with the addition of powder from 2.52 to 2.19 to 2.10. The a value was higher in Kimchi with added Backryeoncho powder in a dose-dependent manner compared with control Kimchi. The cutting force and compression force were higher during the fermentation period, and especially addition of 0.4% powder showed the highest value of hardness. The addition of powder slowed growth of lactic acid bacteria as compared with the control samples. In the sensory evaluation of overall taste, 0.4% addition of powder improved the quality of Kimchi. Therefore, addition of 0.4% Backryeoncho powder apears to be an acceptable approach to enhance the quality of Kimchi without reducing acceptability.
Journal of the korean academy of Pediatric Dentistry
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v.24
no.1
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pp.204-219
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1997
The use of fluoride is one of the most effective methods for caries prevention. Fluoridation of public water supply has been recognized, for many years, as an effective way to reduce dental caries. The fluoride supplement has been recommended when the natural fluoride was unavailable or below the optimal range. However the mechanism of caries prevention by fluoride has not yet been clarified and it is well known that an overdose of fluoride results inacute and chronic toxicity, especially dental fluorosis. Fluoride mouthrinsing solution is widely used in dentistry due to its effectiveness in carrying anticariogenic action. Understanding the effects of fluoride mouthrinsing solution on human gingival fibroblasts will provide the safety rationale for its use during the caries preventive therapy. The purpose of this study was to evaluate the cytotoxic effect of fluoride mouthrinsing solution on the human gingival fibroblast in vitro. The human gingival fibroblasts were cultured from healthy gingiva on the extracted deciduous teeth of children. Cells were inoculated into a 24-well plate with $1{\times}10^4cells/well$ of medium at $37^{\circ}C$, 100% humidity, 5% $CO_2$ incubator for 24 hours. And the cells were counted by using the hemocytometer at each designed study. Human gingival fibroblasts were cultured in growth medium after one minute application range of 0.02%-0.2% NaF solution and 0.1% $SnF_2$ solution. The cells used in this study were between fifth to eighth passage number. The cell morphology was examined by inverted microscope and cell proliferation was measured by incorporating $[^3H]$-thymidine into DNA. DNA synthesis by human gingival fibroblasts was assessed by $[^3H]$-thymidine uptake assays while the cell activity was measured by MTT assay. Each concentrated fluoride mouthrinsing solution was estimated for its biocompatability with fibroblasts by the tissue culture technique. The results of this study were as follows : 1. It was observed that at 0.05%, 0.2% NaF mouthrinsing solution the cytoplasmic processes became globular. When 0.1% $SnF_2$ mouthrinsing solution was applied, the cytoplasmic process and cell morphology were disappeared. 2. DNA synthetic activity was reduced regardless of the concentration of the fluoride mouthrinsing solution. However, the result is statistically insignificant except 0.1% $SnF_2$ mouthrinsing solution(p<0.05). 3. Our results indicate that 0.02%, 0.05% concentrations of NaF mouthrinsing solution caused minimal cytotoxicity. But 0.2% NaF and 0.1% $SnF_2$ concentration were a significant difference between the cell activity in the experimental group and control group (p<0.05). 4. After appling 0.05% & 0.02% NaF fluoride mouthrinsing solution, cell activity was restored to the control groups level according to incubating time. The results suggest that direct exposure to fluoride solution inhibits gingival fibroblast activity. Therefore, for the most effective use of fluoride use, lowering the concentration of fluoride mouthrinsing is advisable because it maintains biocompatability and free ion in the oral fluid.
Helicobacter pylori is a spiral-shaped, microaerophilic human gastric pathogen causing chronic-active gastritis in association with duodenal ulcer and gastric cancer. To investigate the possibility of H. pylori outer membrane proteins (OMPS) as the oral vaccine antigens, sarcosine-insoluble outer membrane fraction has been prepared from H. pylori NCTC 11637. The major OMPs having apparent molecular masses of 62 kDa, 54 kDa and 33 kDa were detected by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), which were identified as urease B subunit (UreB), heat shock protein (Hsp54 kDa) and urease A subunit (UreA), respectively. Minor protein bands of 57 kDa, 52 kDa, 40 kDa, 36 kDa and 31 kDa were also observed. The antigenicity of H. pylori OMPs and antigenic cross-reactivity among the strains were determined by immunoblot analysis using anti-H. pylori OMPs antisera or intestinal lavage solutions. The results showed that UreB, Hsp54 kDa, UreA and 40 kDa proteins vigorously stimulated mucosal immune response rather than systemic immunity. From this results, these proteins seemed to be useful as the antigen candidates for the oral vaccine. The immunoblotting results with surface proteins from eight isolated H. pylori strains were similar to that of H. pylori NCTC 11637. The IgA which had been arised from oral administration of H. pylori OMPs, was able to bind H. pylori whole-cells.
Improvement of L-asparate ${\beta}-decarboxylase$ production from Pseudomonas dacunhae ATCC 21192 was attempted by optimizing fermentation conditions. Optimum carbon and nitrogen sources for cell growth and enzyme production were determined. L-Glutamate (2%) was the most suitable carbon source, and D-glucose, D-glycerol and fumarate repressed enzyme production. Yeast extract (2%) was the most effective as nitrogen source. A slight change of pH to 6.5 from medium pH resulted in a meaningful increase in the production of enzyme. The production of the enzyme was highly improved by using 2% yeast extract and 2% L-glutamate in culture media. Maximum L-asparate ${\beta}-decarboxylase$ activity reached up to over 24 U/mL-broth by 15 h flask fermentation.
This study was to examine whether the in vitro friability, motility and intact acrosome of frozen-thawed bovine and human sperm can be improved by adding Pentoxifylline (PF) or Fertilization Promoting Peptide (FPP). Human semen was frozen ultra-rapidly using Test yolk-buffer (TYB) freezing medium. Additive (PF, FPP) effects in frozen-thawed bovine and human sperm were analyzed by microscopic count for sperm motility and coomassie brilliant blue staining method f3r sperm acrosome intact. The in vitro motility of frozen-thawed bovine sperm with 5 mM PF treatment group (50.0%) was significantly higher than that of control (34.0%) (P<0.05). In the frozen-thawed bovine sperm was examined, the intact acrosome rate of 50 nM FPP treatment (49.0%) was significantly higher than those of control (30.0%) and 25 nM FPP (38.0%) treatment groups (P<0.01). In human semen, when in vitro motility of sperm with PF addition prior to freezing was examined, the result of 5 mM treatment group (51.0%) was significantly higher than those of control and 2.5 mM treatment group (39.0, 40.0%) (P<0.01). In addition, 50 nM (75.5%) FPP adding in all treatment procedures for human semen freezing (before freezing, freezing and after thawing) was significant effect on maintenance of the sperm intact acrosome percentage (control: 45.0; 25 nM: 53.0; 100 nM: 68.0%) (P<0.01). Also, the intact acrosome rate of human sperm with FPP (65.0%) was significantly higher than that with PF (43.0%) (P<0.05), although sperm motility was slightly higher in PF treatment group. These results suggest that improved sperm motility and intact acrosome of frozen thawed bovine and human sperm can be obtained by addition of PF or FPP, and that the enhanced in vitro viability, motility and intact acrosome can be obtained by addition of FPP in all semen freezing procedures.
Highly toxic gases such as hydrogen sulfide ($H_2S$), carbon dioxide ($CO_2$), and ammonia ($NH_3$) are generated by both nature and human activities and affect human health. In this research, activated carbon combined with $Ca(OH)_2$ and $CaCO_3$ (AC-CO and AC-CC, respectively) were fabricated and applied in absorbing toxic gases from air pollutants. Activated charcoal powder was compressed in the form of pellets and used in the designated conditions. The optimum operating conditions and material properties, such as adsorption capacity, effect of weight ratio of the mixture, and hardness, have been investigated after combining with the calcium derivative. The good performance exhibited in this study suggests that this material is expected to be an effective and economically viable adsorbent for $NH_3$, $CO_2$, and $H_2S$ removal from the air system.
The purpose of this study was to investigate the quality characteristics and antioxidative effect of Dasik prepared with Lycii fructus extract(LD). LD were added in ratios (w/w) of 0(C), 5(LD1), 10(LD2), and 15%(LD3), and then proximate compositions, physicochemical properties, sensory evaluations and antioxidative effect of the Dasik were measured. LD1~LD3 samples showed higher contents of moisture, crude lipid, crude protein and crude ash as well as pH and sugar content compared to control (p<0.001). The total phenol content of LD samples (LD1, 31.5% ; LD2, 66.1% ; and LD3, 69.7%) was higher than that of control (4.71%). DPPH radical scavenging activity of LD3 showed the highest activity of 65.3%, whereas that of control (27.0%) was the lowest among samples. Additionally, ABTS radical scavenging activity of LD samples was higher than that of control (C, 38.9% ; LD1, 53.6%; LD2, 80.5% ; and LD3, 83.3%). The results of texture analysis on all samples showed that control had the highest but LD3 was the lowest (p<0.001). In sensory evaluation, the scores of appearance, flavor, taste and overall preference for LD2 was significantly higher than the samples(C, LD1, and LD2). Form the findings, this study suggests that 10% addition of LD was effective for preparation of Dasik in the aspects of the consumer acceptability and functionality.
Objectives : This study was focused to investigate the toxicity of Saussurea lappa (SL) extracts in HL-60 cells and human lymphocytes. We also examined the differentiation effect of SL against leukemia cells. Methods : For examining the toxicity of SL, cytokinesis-block micronucleus (CBMN) assay and single cell gel eletrophoresis (SCGE) assay were used in present study. The cell differentiation effect of SL was evaluated by nitroblue tetrazolium (NBT) reduction assay. Results : The inhibition of cell growth in HL-60 cells was observed in a dose-dependant manner after SL treatment for 24 h. According to SCGE assay, HL-60 cells treated with SL increased DNA damage at $10{\mu}g/m{\ell}$, while DNA damage was induced by 0.1, 1, $10{\mu}g/m{\ell}$ concentration of SL in human lymphocytes. Our results indicated that SL have no genotoxic effect in HL-60 cells and human lymphocytes. Additionally, the differentiation effect was induced in $1{\mu}g/m{\ell}$ SL-treated HL-60 cells. Conclusions : From above results it is suggested that SL could be beneficial for the preparation of the useful agent for treating leukemia.
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