Choi, Hyo Jin;Hwang, Sang Youn;Jang, Dae Ho;Cho, Hyung Min;Kang, Jung Hye;Seong, Gi Hun;Choo, Jae Bum;Lee, Eun Kyu
Korean Chemical Engineering Research
/
v.44
no.1
/
pp.65-72
/
2006
Recent technical advances in the biorecognition engineering and the microparticle fabrication may enable us to develop the single step purification using magnetic particle, because of its simplicity, efficacy, ease of automation, and process economics. In this study, we used commercial magnetic particles from Seradyn, Inc. (Indianapolis, USA). It was ca. 2.8 micron in diameter, consisted of polystyrene core and magnetite coating, and its surface had carboxyl groups. The model, capture protein was IgG and anti-IgG was used as the ligand molecule. We studied the different surfaces ('nude', ester-activated, and anti-IgG coated) for their biorecognition of IgG. At a high pH condition, we could reduce non-specific binding. Also anti-IgG immobilized magnetic particle could capture IgG more selectively. We attempted 'oriented immobilization' of anti-IgG, in which the polysaccharides moiety near the C-terminus was selectively oxidized and linked to the hydrazine-coated MP, to improve the efficacy of biorecognitive binding. Using this method, the IgG capturing ability was improved by ca. 2 fold. From the binary mixture of the IgG-insulin, IgG could be more selectively captured. In summary, the oriented immobilization of oxidized anti-IgG proved to be as effective as the streptavidin-biotin system and yet simpler and cost-effective. This immobilization method can find its applications in protein biochips and biotargeting.
Kang, Kyung Hun;Park, Si Young;Kwon, Ki Han;Lim, Heekyung;Kim, Sung Hyun;Kim, Jeong Gyun;Chung, Mi Ja
Journal of the Korean Society of Food Science and Nutrition
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v.44
no.10
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pp.1458-1469
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2015
This study was carried out to investigate the nitrite scavenging activities (NSA) of nine kinds of wild vegetables in a $NaNO_2$ model system and nitrite of Chinese cabbage as well as the inhibitory effect of kimchi containing a mixture of wild vegetables (MWV) with nitrite scavenging activity on brain neuronal cell death. NSA was higher at pH 1.2 than pH 4.2 in all samples. NSA of extracts from sprouts of Oenothera laciniata and Aster scaber (AS) was above 90% at pH 1.2. AS, Codonopsis lanceolate (CL), Adenophora triphylla (AT), Platycodon grandiflorum (PG), and Taraxacum officinale (TO) extracts showed significantly higher levels of NSA than those from other extracts at pH 4.2. CL, AT, PG, and TO extracts showed high NSA on nitrite of Chinese cabbage. In addition, the effects of MWV on antioxidant and brain neuronal cell death induced by oxidative stress were investigated in human brain neuroblastoma SK-N-SH cells. MWV extract attenuated $H_2O_2$-induced cell death and reactive oxygen species (ROS) generation in SK-N-SH cells. MWV extract showed significantly higher DPPH radical scavenger activity when compared to normal kimchi extract. MWV extract showed an inhibitory effect on brain neuronal cell death against oxidative stress by antioxidant activities.
Song, Ji-Hye;Hwang, Dong Hyeon;Oh, Doo-Byoung;Rhee, Sang Ki;Kwon, Ohsuk
Microbiology and Biotechnology Letters
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v.41
no.1
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pp.17-25
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2013
The thermotolerant methylotrophic yeast Hansenula polymorpha is an attractive model organism for various fundamental studies, such as the genetic control of enzymes involved in methanol metabolism, peroxisome biogenesis, nitrate assimilation, and resistance to heavy metals and oxidative stresses. In addition, H. polymorpha has been highlighted as a promising recombinant protein expression host, especially due to the availability of strong and tightly regulatable promoters. In this study, we investigated the possibility of employing human serum albumin (HSA) as the fusion tag for the secretory expression of heterologous proteins in H. polymorpha. A set of four expression cassettes, which contained the methanol oxidase (MOX) promoter, translational HSA fusion tag, and the terminator of MOX, were constructed. The expression cassettes were also designed to contain sequences for accessory elements including His8-tag, $2{\times}(Gly_4Ser_1)$ linkers, tobacco etch virus protease recognition sites (Tev), multi-cloning sites, and strep-tags. To determine the effects of the size of the HSA fusion tag on the secretory expression of the target protein, each cassette contained the HSA gene fragment truncated at a specific position based on its domain structure. By using the Green fluorescence protein gene as the reporter, the properties of each expression cassette were compared in various conditions. Our results suggest that the translational HSA fusion tag is an efficient tool for the secretory expression of recombinant proteins in H. polymorpha.
Recent underground common utility tunnels are underground facilities for jointly accommodating more than 2 kinds of air-conditioning and heating facilities, vacuum dust collector, information processing cables as well as electricity, telecommunications, waterworks, city gas, sewerage system required when citizens live their daily lives and facilities responsible for the central function of the country but it is difficult to cope with fire accidents quickly and hard to enter into common utility tunnels to extinguish a fire due to toxic gases and smoke generated when various cables are burnt. Thus, in the event of a fire, not only the nerve center of the country is paralyzed such as significant property damage and loss of communication etc. but citizen inconveniences are caused. Therefore, noticing that most fires break out by a short circuit due to electrical works and degradation contact due to combustible cables as the main causes of fires in domestic and foreign common utility tunnels fire cases that have occurred so far, the purpose of this paper is to scientifically analyze the behavior of a fire by producing the model of actual common utility tunnels and reproducing the fire. A fire experiment was conducted in a state that line type fixed temperature detector, fire door, connection deluge set and ventilation equipment are installed in underground common utility tunnels and transmission power distribution cables are coated with fire proof paints in a certain section and heating pipes are fire proof covered. As a result, in the case of Type II, the maximum temperature was measured as $932^{\circ}C$ and line type fixed temperature detector displayed the fire location exactly in the receiver at a constant temperature. And transmission power distribution cables painted with fire proof paints in a certain section, the case of Type III, were found not to be fire resistant and fire proof covered heating pipes to be fire resistant for about 30 minutes. Also, fire simulation was carried out by entering fire load during a real fire test and as a result, the maximum temperature is $943^{\circ}C$, almost identical with $932^{\circ}C$ during a real fire test. Therefore, it is considered that fire behaviour can be predicted by conducting fire simulation only with common utility tunnels fire load and result values of heat release rate, height of the smoke layer, concentration of O2, CO, CO2 etc. obtained by simulation are determined to be applied as the values during a real fire experiment. In the future, it is expected that more reliable information on domestic underground common utility tunnels fire accidents can be provided and it will contribute to construction and maintenance repair effectively and systematically by analyzing and accumulating experimental data on domestic underground common utility tunnels fire accidents built in this study and fire cases continuously every year and complementing laws and regulations and administration manuals etc.
We investigated the binding properties of $(^3H)$ QNB and $(^3H)$ NMS to mAchR to elucidate the characterstics of mAchR in rat brain by using two different preparations (homogemates & intact brain cell aggregates). The binding properties of both ligands demonstrated high affinity and saturability in both experiments, however $(^3H)$ QNB showed a significantly higher maximal binding capacity than tha ot $(^3H)$ NMS 1. In rat brain homogenates; Displacement of both lignands with several mAchR antagonists resulted in competition curves in accoradnce with the law of massaction for QNB, atropine & scopolamine in thie preparation, also a similar profile was found for the quaternary ammonium analogs of atropine & scopolamine (methyl atropine & methylscopolamine) when $(^3H)$ NMS was used to label the receptors in rat brain. But when these hydrophillic antagonists were used to displace $(^3H)$ QNB, they showed interaction with high- and low-affinity binding sites in brain homogenates. Pirenzepine, the nonclassical mAchR antagonist, was able to displace both ligands from binding sites in this preparation. 2. In intact rat brain cell aggregates; Intact bain cell aggregates were used to elucidate the binding characteristics of $(^3H)$ NMS to mAchR in rat. The magnitude of binding of this ligand was related linearly to the amount of cell protein in the binding assay with a high ratio of total to nonspecific binding. mAchR antagonists displaced specific $(^3H)$ NMS binding according to the law of mass-action, while it was possible to resolve displacement curves using mAchR agonist into high-& low-affinity component. 3. Our results indicate that more hydrophilic receptor ligand $(^3H)$ QNB, displacement experiments in both tissues demonstrated that the lipid solubility of a particulr mAchR ligand might play an important role in determining its profile of binding to the mAchR, and the concentrations of mAchR in rat brain are both on the cell surface (membrane-bound receptor) and in the intracelluar membrane (intermembrane-bound receptor). 4. The results are discussed in terms of the usefulness of dissociated intact rat brain cells in studying mAchR in central nervous system.
Emission source term is one of the strong controlling factors for the air quality simulation capability, particularly over the urban area. Ulsan is an industrial area and frequently required to simulate for environmental assessment. In this study, two CAPSS (Clean Air Policy Support System) emission data; CAPSS-2003 and CAPSS-2010 in Ulsan, were employed as an input data for WRF-CMAQ air quality model for emission assessment. The simulated results were compared with observations for the local emission dominant synoptic conditions which had negative vorticities and lower geostrophic wind speed at 850hPa weather maps. The measurements of CO, $NO_2$, $SO_2$ and $PM_{10}$ concentrations were compared with simulations and the 'scaling factors' of emissions for CO, $NO_2$, $SO_2$, and $PM_{10}$ were suggested in in aggregative and quantitative manner. The results showed that CAPSS-2003 showed no critical discrepancies of CO and $NO_2$ observations with simulations, while $SO_2$ was overestimated by a factor of more than 12, while $PM_{10}$ was underestimated by a factor of more than 20 times. However, CAPSS-2010 case showed that $SO_2$ and $PM_{10}$ emission were much more improved than CAPSS-2003. However, $SO_2$ was still overestimated by a factor of more than 2, and $PM_{10}$ underestimated by a factor of 5, while there was no significant improvement for CO and $NO_2$ emission. The estimated factors identified in this study can be used as'scaling factors'for optimizing the emissions of air pollutants, particularly $SO_2$ and $PM_{10}$ for the realistic air quality simulation in Ulsan.
The aim of this study was to investigate whether fourier transform infrared (FT-IR) spectroscopy can be applied to simultaneous determination of fatty acids contents in different soybean cultivars. Total 153 lines of soybean (Glycine max Merrill) were examined by FT-IR spectroscopy. Quantification of fatty acids from the soybean lines was confirmed by quantitative gas chromatography (GC) analysis. The quantitative spectral variation among different soybean lines was observed in the amide bond region ($1,700{\sim}1,500cm^{-1}$), phosphodiester groups ($1,500{\sim}1,300cm^{-1}$) and sugar region ($1,200{\sim}1,000cm^{-1}$) of FT-IR spectra. The quantitative prediction modeling of 5 individual fatty acids contents (palmitic acid, stearic acid, oleic acid, linoleic acid, linolenic acid) from soybean lines were established using partial least square regression algorithm from FT-IR spectra. In cross validation, there were high correlations ($R^2{\geq}0.97$) between predicted content of 5 individual fatty acids by PLS regression modeling from FT-IR spectra and measured content by GC. In external validation, palmitic acid ($R^2=0.8002$), oleic acid ($R^2=0.8909$) and linoleic acid ($R^2=0.815$) were predicted with good accuracy, while prediction for stearic acid ($R^2=0.4598$), linolenic acid ($R^2=0.6868$) had relatively lower accuracy. These results clearly show that FT-IR spectra combined with multivariate analysis can be used to accurately predict fatty acids contents in soybean lines. Therefore, we suggest that the PLS prediction system for fatty acid contents using FT-IR analysis could be applied as a rapid and high throughput screening tool for the breeding for modified Fatty acid composition in soybean and contribute to accelerating the conventional breeding.
Purpose: The aim of this study is to identify clinical usefulness of Wide Beam Reconstruction (WBR) which is called Xpress.cardiac$^{TM}$ to confirm the agreement between segmental perfusion and regional wall motion in myocardium compared to conventional OSEM method. Materials and Methods: Subjects were separated two groups. First group was composed of 20 normal control group. Second group was composed of 10 patients (abnormal group) who had coronary artery disease. Subjects underwent myocardial perfusion SPECT ($^{201}Tl$ rest and $^{99m}Tc$-MIBI stress). Image acquisition and reconstruction were that rest stage was each step per 30, 15 seconds and stress stage was each step per 25, 13 seconds, OSEM and WBR methods were applied. Segmental perfusion and regional wall motion were applied 20-segment model of QPS, QGS algorithm in AutoQuant. Status of perfusion was composed of 5 point scoring system (0=normal, 1=mild, 2=moderate, 3=severe hypokinesia, 4=dyskinesia). Status of regional wall motion was also composed of 5 point scoring (0=normal, 1=mild, 2=moderate, 3=severe hypokinesia, 4=dyskinesia). We evaluated the agreement between conventional OSEM and WBR through automatic quantification value. Results: The agreement of rest segmental perfusion between conventional OSEM and WBR in normal patients was 99% (396/400, k=0.662, p<0.0001) and one of rest regional wall motion was 83.8% (335/400, k=0.283), the agreement of stress segmental perfusion was 95.8%(383/400, k=0.656), one of stress regional wall motion was 87.3% (349/400, k=0.390). The match rate of rest segmental perfusion in abnormal patients was 83% (166/200, k=0.605, p<0.0001) and one of rest regional wall motion was 55.5% (111/200, k=0.385), the agreement of stress segmental perfusion was 79.5% (159/200, k=0.682), one of stress regional wall motion was 63.5% (127/200, k=0.486). Conclusion: Compared to conventional OSEM, WBR method had a good agreement of segmental perfusion in myocardium in normal and abnormal groups. However regional wall motion showed meaningful low agreement. Although WBR offers high resolution and contrast ratio, it is not useful method for gated myocardial perfusion SPECT.
The principal objective of the current study was to isolate a purpurogallin derivative as an oxidation product from gallic acid, in an effort to assess the anti-inflammatory effects of this compound. Purpurogallin derivative is known to be the one of the oxidation products of gallic acid. This compound has been identified as a minor chemical component in fermented tea products. It has been previously demonstrated that theaflavins, the oxidation products of catechins found in fermented tea products, exert profound antioxidant and anti-inflammatory effects. However, the biological activities of a minor chemical component in fermented teas have yet to be evaluated. Purpurogallin carboxylic acid (PCA) was identified as a major oxidation product of gallic acid from a peroxidase/hydrogen peroxide oxidation model system. The identity of the PCA was verified by $^{1}H$ NMR, $^{13}C$ NMR and MS techniques. PCA treatment significantly suppressed the generation of pro-inflammatory mediators including nitric oxide and IL-6 in lipopolysaccharide (LPS)-stimulated RAW264.7 murine macrophages. According to the nitrite assay, PCA 100, 75, and $50{\mu}g/mL$ treatment dose-dependently inhibited NO production by 57.6, 41.5, and 21.8%, respectively, in LPS-stimulated RAW264.7 murine macrophage cells. Moreover, IL-6 production was inhibited to a significant degree with PCA treatment of 100 and $75{\mu}g/mL$ at 43.1 and 23.9%, respectively. PCA treatment also significantly suppressed $PGE_2$ production at levels of 100 and $75{\mu}g/mL$. These results showed that PCA exerts inhibitory effects on the production of inflammatory mediators.
The purpose of this study was to examine the presence of Bacillus cereus, aerobic bacteria and coliforms in the raw material of infant formulas and investigate the manufacturing process in terms of microbial safety. Among ten kinds of raw infant formula material samples (n=20), Bacillus cereus appeared in two (n=4). Aerobic bacteria were not detected in raw infant formula material or maximum 4.15 log CFU/g. Eleven species of aerobic bacteria were isolated and 76% of them were Sphingomonas paucimobilis, Pseudomonas fluorescens, Rhizobium radiobactor, or Stenotrophomonas maltophilia. A Pearson's correlation analysis revealed that the most influential factors for detecting Bacillus cereus were aerobic bacteria and coliforms. In other words, when the measured values of aerobic bacteria and coliforms were higher, the possibility that Bacillus cereus would appear increased. In a regression model to predict Bacillus cereus, the rate of appearance was correlated with aerobic bacteria and coliforms, and its contribution rate for effectiveness was 86%. Improving microbial quality control by pasteurization, spray dry, popping and extrusion resulted in a decrease in the numbers of Bacillus cereus, aerobic bacteria and coliforms in the raw materials. The results suggest that a hazard analysis and critical control point system might be effective for reducing microbiological contamination.
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