• Asian-Australasian Journal of Animal Sciences
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• v.29 no.1
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• pp.126-133
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• 2016

A gene from Actinomyces sp. Korean native goat (KNG) 40 that encodes an endo-${\beta}$-1,4-glucanase, EG1, was cloned and expressed in Escherichia coli (E. coli) $DH5{\alpha}$. Recombinant plasmid DNA from a positive clone with a 3.2 kb insert hydrolyzing carboxyl methyl-cellulose (CMC) was designated as pDS3. The entire nucleotide sequence was determined, and an open-reading frame (ORF) was deduced. The ORF encodes a polypeptide of 684 amino acids. The recombinant EG1 produced in E. coli $DH5{\alpha}$ harboring pDS3 was purified in one step using affinity chromatography on crystalline cellulose and characterized. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis/zymogram analysis of the purified enzyme revealed two protein bands of 57.1 and 54.1 kDa. The amino terminal sequences of these two bands matched those of the deduced ones, starting from residue 166 and 208, respectively. Putative signal sequences, a Shine.Dalgarno-type ribosomal binding site, and promoter sequences related to the consensus sequences were deduced. EG1 has a typical tripartite structure of cellulase, a catalytic domain, a serine-rich linker region, and a cellulose-binding domain. The optimal temperature for the activity of the purified enzyme was $55^{\circ}C$, but it retained over 90% of maximum activity in a broad temperature range ($40^{\circ}C$ to $60^{\circ}C$). The optimal pH for the enzyme activity was 6.0. Kinetic parameters, $K_m$ and $V_{max}$ of rEG1 were 0.39% CMC and 143 U/mg, respectively.

• Journal of Microbiology and Biotechnology
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• v.27 no.11
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• pp.2044-2051
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• 2017

The main pathological hallmark of Alzheimer's disease is the deposition of amyloid-beta ($A{\beta}$) peptides in the brain. $A{\beta}$ has been widely used to mimic several aspects of Alzheimer's disease. However, several characteristics of amyloid-induced Alzheimer's disease pathology are not well established, especially in mice. The present study aimed to develop a new Alzheimer's disease model by investigating how $A{\beta}$ can be effectively aggregated using prokaryotes and eukaryotes. To express the $A{\beta}42$ complex in HEK293 cells, we cloned the $A{\beta}42$ region in a tandem repeat and incorporated the resulting construct into a eukaryotic expression vector. Following transfection into HEK293 cells via lipofection, cell viability assay and western blotting analysis revealed that exogenous $A{\beta}42$ can induce cell death and apoptosis. In addition, recombinant His-tagged $A{\beta}42$ was successfully expressed in Escherichia coli BL21 (DE3) and not only readily formed $A{\beta}$ complexes, but also inhibited the proliferation of SH-SY5Y cells and E. coli. For in vivo testing, recombinant His-tagged $A{\beta}42$ solution ($3{\mu}g/{\mu}l$ in $1{\times}PBS$ containing $1mM\;Ni^{2+}$) was injected stereotaxically into the left and right lateral ventricles of the brains of C57BL/6J mice (n = 8). Control mice were injected with $1{\times}PBS$ containing $1mM\;Ni^{2+}$ following the same procedure. Ten days after the sample injection, the Morris water maze test confirmed that exogenous $A{\beta}$ caused an increase in memory loss. These findings demonstrated that $Ni^{2+}$ is capable of complexing the 50-kDa amyloid and that intracerebroventricular injection of $A{\beta}42$ can lead to cognitive impairment, thereby providing improved Alzheimer's disease models.

• Proceedings of the Korea Society of Environmental Toocicology Conference
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• 2003.10a
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• pp.57-69
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• 2003

It is well known that many pesticides possess hormonal activity, and affect the developments of wildlife and mammals including human. Currently, pyrethroid insecticides are in worldwide use to control in and outdoor pests, providing potential far environmental exposure. Hormonal activities of these pyrethroid insecticides, however, have been little studied, and the developmental effects of them were no reported. Therefore, we firstly examined the potential estrogenic activities of some pyrethroid insecticides (permethrin, cypermethrin, tetramethrin, deltamethrin, sumithrin, fenvalerate and bioallethrin) by immature rat uterotrophic assay, luciferase reporter gene assay and Calbindin-D$\sub$9k/ (CaBP-9k) gene expression assay. Uterine wet weights were increased by permethrin and the permethrin-induced weights were inhibited by ICI 182780 in the uterolrophic assay. On the other hand tetramethrin significantly reduced uterine and vaginal wet weights, and also inhibited the E2-induced weight increases at all doses tested. Cypermethrin and sumithrin had a tendency to increase uterine weights, although not statistically significant. Permethrin and cypermethrin dose-dependently increased the luciferase activity in reporter gene assay. Northern blot analysis showed that permethrin induced CaBP-9k mRNA expression whereas tetramethrin inhibted. Subsequent studies were conducted to investigate the possible developmental effects of four pyrethroid insecricides (permethrin, cypermethrin, sumithrin and teramethrin). Either diethlbestrol (DES) or 17${\beta}$ -estradiol (E2) was used as a reference control in this study. Pyrethroid insecticides were administered to Sprague Dawley rats via subcutaneous injection at 6 to 18 days of gestation or 1 to 5 days after birth. In utero treatment of permethrin (10mg/kg/day) in female rat resulted in significant increases in uterine and ovarian weights while significant decreases in serum E2 concentration, uterine and ovarian ER${\alpha}$ mRNA levels. Sumithrin and permethrin led to acceleration in vaginal opening of female rat, while delay in preputial separation of male after neonatal treatment. Anogenital distances of PND 18 were significantly reduced in sumthrin-treated, and permerhrin-treated male rats after neonatal treatment. All the pyrethroid insecticides tested caused significant increases in uterine weights on PND 18, while significant reductions in the first diestrus phase when neonataly treated. In addition, exposure to pyrethroids in neonatal period led to significant reduction in relative brain weight in female rat on PND 18, but its weight was recovered in diestrus phase. In summary, Our experimental data demonstrate the possibilities of developmental effects of pyrethroid insecticides via estrogenic or antiestrogenic activity.

• Nutrition Research and Practice
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• v.4 no.1
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• pp.3-10
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• 2010

This study was performed to investigate the effect of desalinated underground seawater (named as 'magma seawater', MSW) of Jeju Island in Korea on lipid metabolism and antioxidant activity. MSW was collected from underground of Han-Dong in Jeju Island, and freely given to high fat diet (HFD)-fed C57BL/6 mice for 10 weeks. Although there were no significant differences in the body weight changes and plasma lipid levels, hepatic triglyceride levels were significantly lower in the MSW group than in the normal tap water (TW)-drunken control group. Furthermore, the activity of fatty acid synthase (FAS) was significantly decreased and carnitine palmitoyltransferase (CPT) activity was increased in MSW group compared to TW group. Similarly, real-time PCR analysis revealed that mRNA expressions of lipogenic genes were lowered in MSW groups compared to the control group. In a morphometric observation on the liver tissue, accumulation of fats was remarkably reduced in MSW group. Meanwhile, in vitro assay, tree radical scavenging activity measured by using diphenylpicrylhydrazyl (DPPH) was increased in MSW group. The 2'-7'-dichlorofluorescein diacetate (DCF-DA) staining followed with fluorescent microscopy showed a low intensity of fluorescence in MSW-treated HepG2 cells, compared to TW-treated HepG2 cells, which indicated that the production of reactive oxygen species by tert-butyl hydroperoxide (t-BHP) in HepG2 cells was decreased by MSW treatment. The antioxidant effect of MSW on t-BHP-induced oxidative stress in HepG2 cells was supported by the increased activities of intracellular antioxidant enzymes such as catalase and glutathione reductase. From these results, we speculate that MSW has an inhibitory effect on lipogenesis in liver and might play a protective role against cell damage by t-BHP-induced oxidative stress.

• Herbal Formula Science
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• v.24 no.2
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• pp.108-123
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• 2016

Objectives : This comparative study was to investigate on anti-obesity effects of Ephedrae Herba and Cyperi rhizoma in high fat diet(HFD) fed mice. Methods : Male C57BL/6J mice were fed a normal diet(normal group, N), high fat 45 cal% diet[HFD, control group, C), HFD with Ephedrae Herba(EH group) and Cyperi rhizoma(CR group) extracts fed for 5 weeks. We were observed as follows : changes of body weight, amount of diet intake, weight of total visceral fats, levels of obesity-related hormones and blood lipids. Results : The change of body weight after EH and CR oral administration significantly more decreased in EH group than that of control group. The FFR(Food Efficiency Ratio) was decreased in EH group, but more increased in CR group than that of control group. The weight of periepididymal and perirenal fats were significantly decreased in EH and CR groups compared to the control group. The levels of serum leptin and insulin were significantly decreased in EH group, and the level of serum adiponectin was increased in EH group compared to control group. The levels of serum triglyceride and total cholesterol were significantly decreased in EH and CR groups, and HDL-cholesterol levels was significantly increased in EH group compared to control group. Conversely in CR group, its values showed the opposite effect. The staining density of lipid droplets within the hepatocytes was widely distributed in CR and control groups, but in EH group, its density was weakly stained. Conclusions : These experimental results suggest that Ephedrae Herba shows conspicuous anti-obesity effect, and Cyperi rhizoma shows weak anti-obesity effect.

• Clinical and Experimental Pediatrics
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• v.63 no.5
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• pp.189-194
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• 2020

Background: The decision to use transannular patching (TAP) during tetralogy of Fallot (TOF) repair depends on the pulmonary valve annulus size; the z score of the pulmonary annulus is the most commonly used predictor. However, definitive results are not obtained with z scores as different z score data sets are used for different parameters. Purpose: This study aimed to identify the echocardiographic and other key factors that warranted a change in the surgical method during TOF surgery. Methods: Sixty-two patients were enrolled and divided into a pulmonary valve (PV) preservation group and a TAP group. Their medical records were reviewed. Results: The z score for PV annulus (PVA), ratio of the PVA to aortic annulus size, and ratio of PVA to descending aorta (DAO) size were significantly different between the PV preservation and TAP groups (-1.72±1.52 vs. -3.07±1.94, P=0.004; 0.62±0.12 vs. 0.50±0.14, P=0.002; and 1.32±0.32 vs. 1.07±0.36, P=0.008, respectively). For TAP repair, the PVA z score had a sensitivity of 65.4% and specificity of 73.1%, ratio of PVA to aortic annulus size had a sensitivity of 73.1% and specificity of 65.4%, and ratio of PVA to DAO size had a sensitivity of 69.2% and specificity of 57.7%. The TAP group showed more monocuspid PVs (P=0.011), while the PV preservation group showed more tricuspid PVs (P=0.027). Commissurotomy was more frequently performed in the PV preservation group than in the TAP group (P=0.001). Of patients with commissurotomy, 58% showed a PV z score<-2. Conclusion: Although various echocardiographic parameters may serve as predictors for determining surgical methods for TOF patients, the PV morphology and tissue characteristics should also be considered.

• Journal of Preventive Medicine and Public Health
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• v.51 no.1
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• pp.33-40
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• 2018

Objectives: The aim of this study was to determine the factors associated with the spatial distribution of the incidence of colorectal cancer (CRC) in the neighborhoods of Tehran, Iran using Bayesian spatial models. Methods: This ecological study was implemented in Tehran on the neighborhood level. Socioeconomic variables, risk factors, and health costs were extracted from the Equity Assessment Study conducted in Tehran. The data on CRC incidence were extracted from the Iranian population-based cancer registry. The $Besag-York-Molli{\acute{e}}$ (BYM) model was used to identify factors associated with the spatial distribution of CRC incidence. The software programs OpenBUGS version 3.2.3, ArcGIS 10.3, and GeoDa were used for the analysis. Results: The Moran index was statistically significant for all the variables studied (p<0.05). The BYM model showed that having a women head of household (median standardized incidence ratio [SIR], 1.63; 95% confidence interval [CI], 1.06 to 2.53), living in a rental house (median SIR, 0.82; 95% CI, 0.71 to 0.96), not consuming milk daily (median SIR, 0.71; 95% CI, 0.55 to 0.94) and having greater household health expenditures (median SIR, 1.34; 95% CI, 1.06 to 1.68) were associated with a statistically significant elevation in the SIR of CRC. The median (interquartile range) and mean (standard deviation) values of the SIR of CRC, with the inclusion of all the variables studied in the model, were 0.57 (1.01) and 1.05 (1.31), respectively. Conclusions: Inequality was found in the spatial distribution of CRC incidence in Tehran on the neighborhood level. Paying attention to this inequality and the factors associated with it may be useful for resource allocation and developing preventive strategies in at-risk areas.

• Journal of Physiology & Pathology in Korean Medicine
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• v.24 no.4
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• pp.616-623
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• 2010

Three herbal formulas (Bangpungtongsung-san, Ohyaksungi-san, and Ojeok-san) for wind-cold and heat pain symptom were applied to investigate the immunological activities on antigen (Ag)-specific or Ag-non-specific immune responses in murine macrophage cell line (RAW 264.7) and ovalbumin (OVA)-immunized mice. This study was carried out in nitric oxide (NO) synthesis in RAW 264.7 cells and cellular proliferation in mouse splenocytes according to three herbal formulas. C57BL/6 mice were immunized intraperitonially with OVA/aluminium ($100\;{\mu}g/200\;{\mu}g$/mouse) on day 1, 8, and 15. Three herbal formulas were administrated to mice orally for 3 weeks from day 1. On day 22, OVA-, lipopolysaccharide (LPS)-, and concanavalin A (Con A)-stimulated splenocyte proliferation and antibodies (OVA-specific antibodies of the IgG, lgG1, and total IgM classes) in plasma were measured. Ohyaksungi-san increased NO synthesis in RAW 264.7 cells. Ojeok-san and Ohyaksungi-san significantly enhanced cellular proliferation by LPS and Con A in splenocytes from OVA-immunized mice (p<0.001). Three herbal formulas for wind-cold and heat pain symptom also significantly enhanced plasma OVA-specific IgG, IgG1, and total IgM levels compared with the OVA/Alum group. These results suggested that three herbal formulas for wind-cold and heat pain symptom could be used as stimulator of immune response.

• Korean Journal of Plant Resources
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• v.28 no.2
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• pp.193-216
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• 2015

This study was carried out to investigate the vascular plants of Mt. Bohyeon (Gyeongbuk). The vascular plants were collected 22 times and summarized as 589 taxa, including 105 families, 327 genera, 522 species, 11 subspecies, 48 varieties and 8 forms respectively. Among them, 13 Korean endemics, 10 red list of vascular plants, 2 endangered plants and 83 specially designated plants by the Ministry of Environment were included respectively. The naturalized plants were 37 taxa and the naturalized ratio was 6.41 of 589 taxa. Usage of 589 taxa were consists of 240 taxa (40.1%) of edible plants, 205 taxa (34.2%) of medicinal plants, 62 taxa (10.3%) of ornamental plants, 213 taxa (35.6%) of pasture plants, 15 taxa (2.5%) of timber plants, 8 taxa (1.3%) of fiberplants, and 3 taxa (0.5%) of industrial plants.

• KSBB Journal
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• v.22 no.1
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• pp.7-15
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• 2007

Extensive research was carried out for inhibition of melanin formation as development of whitening cosmetic ingredients. But degradation of melanin itself was not intensively pursued as development of cosmetics. In this study, novel melanin degradation enzyme was developed and characterized. Also this enzyme production process was optimized and formulation was tried using micro encapsulation technique.