• Korean Journal of Clinical Laboratory Science
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• v.46 no.3
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• pp.99-105
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• 2014

Isolating total DNA from small samples using traditional methods is difficult and inefficient mainly due to loss of DNA during filtration and precipitation. With advances in molecular pathology, DNA extraction from micro-dissected cells has become essential in handling clinical samples. Genomic DNA extraction using small numbers of cells can be very important to successfully PCR amplify DNA from small biopsy specimens. We compared our experimental genomic DNA extraction method (A) with two other commercially available methods: using spin columns (B), and conventional resins (C), and determined the efficacy of DNA extraction from small numbers of cells smeared on a glass slide. Approximately 50, 100, 200, 500 and 1000 cells were isolated from fine needle aspiration biopsy (FNAB) slides aspirated from histologically proven papillary thyroid carcinoma masses. DNA was extracted using the three techniques. After measuring DNA quantity, PCR amplification was performed to detect the ${\beta}$-globin and $BRAF^{V600E}$ gene mutations. DNA extracted by method (A) showed better yield than the other methods in all cell groups. With our method, a suitable amount of genomic DNA to produce amplification was extracted from as few as 50 cells, while more than 100 to 200 cells were required when methods (B) or (C) were applied. Our genomic DNA extraction method provides high quality and improved yields for molecular analysis. It will be especially useful for paucicellular clinical samples which molecular pathologists often confront when handling fine needle aspiration cytology, exfoliative cytology and small biopsy specimens.

• Korean Journal of Food Science and Technology
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• v.12 no.2
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• pp.133-139
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• 1980

The extraction mechanism and extraction conditions of cellular lipid of freeze-dried cells of Rhodotorula rubra YUFE 1526, which was reported as coenzyme Q producing microorganism, were studied. 1. Methyl alcohol was the most appropriate solvent for extraction of cellular lipids and the resulting total lipid was 19.17 weight %. 2. When 1 % (w/v) of freeze-dried cell was extracted by methyl alcohol, the extraction yield was 90.49 % at $50^{\circ}C$ for 2 hr. 3. The diffusivity varied with extraction temperature and the empirical equation was derived as follows : $2.2e^{0-02907T}{\times}10^{-9}\;mm^2/min.$ 4. The diffusivity of cellular lipid of Rhodotorula rubra YUFE 1526 was $9.4{\times}10^{-9}\;mm^{2}/min.$ 5. The extraction model was well fitted on the extraction of cellular lipid.

• Journal of the Korean Applied Science and Technology
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• v.31 no.3
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• pp.367-374
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• 2014

In this study, skin permeation enhancement was confirmed by designing it to have a structure and composition similarity to the intercellular lipids that improve miscibility with skin by cross-linked lipids poloxamer. The cross-linked lipids poloxamer was synthesized and analyzed by 1H NMR that structure dose had conjugated pluronic with ceramide3. Active component is released by modification of liquid crystal structure because PPO part, large-scale molecule block of pluronic, has hydrophobic nature at skin temperature of $35^{\circ}C$. Conjugated pluronic with ceramide3 was synthesized using Pluronic F127 and p-NPC (4-nitrophenyl chloroformate) at room temperature yielded 89%. Pluronic(Ceramide 3-conjugated Pluronic) was synthesized by reaction of p-NP-Pluronic with Ceramide3 and DMAP. The yield was 51%. This cross-linked lipids poloxamer was blended and dissolved at isotropic state with skin surface lipids, phospholipid, ceramide, cholesterol and anhydrous additive solvent. Next step was preceded by ${\alpha}$-Transition at low temperature for making the structure of Meso-Phase Lamella, and non-hydrous skin analogue liquid crystal using thermo-sensitivity smart sensor, lamellar liquid crystal structure through aging time. For confirmation of conjugation thermo-sensitivity smart sensor and non-hydrous skin analogue liquid crystal, structural observation and stability test were performed using XRD(Xray Diffraction), DSC(Differential Scanning Calorimetry), PM (Polarized Microscope) And C-SEM (Cryo-Scanning Electron Microscope). Thermo-sensitivity observation by Franz cell revealed that synthesized smart sensor shown skin permeation effect over 75% than normal liquid crystal. Furthermore, normal non-hydrous skin analogue liquid crystal that not applied smart sensor shown similar results below $35^{\circ}C$ of skin temperature, but its effects has increased more than 30% above $35^{\circ}C$.

• The Korea Journal of Herbology
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• v.31 no.6
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• pp.29-35
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• 2016

Objectives : Blood stasis (BS) is related to be caused by blood circulation and stagnation which are cancer, atherosclerosis and hyperlipidemia in traditional medicine. We extracted 3 kinds of BS formula; Seogakjihwang-tang (SGT), Tonggyuhawlhyul-tang (THT), Hyulbuchukeo-tang (HCT). This study was conducted to investigate whether the 3 kinds of herbal formula extracts have inhibitory efficacy association with anti-adipogenesis. Methods : To investigate the anti-adipogenesis, we used the mouse fibroblast cell line, 3T3-L1 which differentiated into adipocytes in response to insulin, IBMX and dexamethasone (MDI). Cytotoxicity of herbal formula extracts were examined by CCK-8 kit. Intracellular lipid droplets were detected by Oil-Red-O staining. Triglyceride (TG) and leptin were measure using elisa kit. Results : The yield of water extracts was 14.62% (SGT), 21.27% (THT), 20.02% (HCT). Lipid accumulation was reduced significantly by 3 kinds of herbal formula compared to control. Especially, THT and HCT decreased lipid droplet, respectively at all concentration. The TG and leptin were also inhibited by 3 kinds of herbal formula. The IC50 of TG were $280.51{\mu}g/m{\ell}$ (SGT), $52.62{\mu}g/m{\ell}$ (THT), $313.99{\mu}g/m{\ell}$ (HCT). The IC50 of leptin were $348.76{\mu}g/m{\ell}$ (SGT), $164.02{\mu}g/m{\ell}$ (THT), $257.00{\mu}g/m{\ell}$ (HCT). THT was better than other herbal formula on anti-adipogenesis. Conclusion : kinds of herbal formula inhibited adipogenesis in MDI-induced 3T3-L1 adipocytes, as indicated by the significant reduction in TG and leptin concentration without cytotoxicity. Therefore, 3 kinds of herbal formula for BS might act as a therapeutic agent for preventing lipid diseases, such as obesity and atherosclerosis.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2019.10a
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• pp.80-80
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• 2019

Potatoes were first introduced outside the Andes region four centuries ago, and have become an integral part of much of the world's food. Potatoes were first introduced into Europe in the 16th century and Korea in the early 19th century. It is the world's fourth-largest crop, following rice, wheat, and maize. In the nutritional aspects, potatoes contain abundant vitamins and minerals, as well as an assortment of phytochemicals such as carotenoids and natural phenols. Chlorogenic acid constitutes up to 90% of potato natural phenols. Due to the high content of potato functional compounds, it has known that potatoes are effective in the prevention of various human diseases. Recently, potato 'Seohong' was developed by RDA, and it has reported that they have a high-yield and dry matter content. Processing industry of potato generates high amounts of peel as a byproduct. It was reported as a good source of several beneficial functional ingredients including antioxidant effect. This study was conducted to enhance the utilization of the peel of Potato 'Seohong'. The anti-inflammatory effects on solvent fraction was evaluated. The anti-inflammatory activities of Ethylacetate fraction was evaluated for inhibitory activities against lipopolysacchride (LPS) induced nitric oxide (NO) in RAW264.7 cell lines. The fraction inhibitory activity for tests with $IC_{50}$ values showed in the ranges of $50{\mu}g/ml$. This result revealed that n-butanol fraction of 'Seohong's peel is expected to be good candidate for development into source of anti-inflammatory agent.

• Geomechanics and Engineering
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• v.6 no.2
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• pp.173-194
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• 2014

Laboratory and field data showed that deep mixed (DM) columns accelerated the rate of consolidation of the soft foundations. Most analyses of consolidation of DM column-improved foundations so far have been based on the elastic theory. In reality, the DM columns may yield due to the stress concentration from the soft soil and its limited strength. The influence of column yielding on the degree of consolidation of the soft foundation improved by DM columns has not been well investigated. A three-dimensional mechanically and hydraulically-coupled numerical method was adopted in this study to investigate the degree of consolidation of the DM column foundation considering column yielding. A unit cell model was used, in which the soil was modeled as a linearly elastic material. For a comparison purpose, the DM column was modeled as an elastic or elastic-plastic material. This study examined the aspects of stress transfer, settlement, and degree of consolidation of the foundations without or with the consideration of the yielding of the DM column. A parametric study was conducted to investigate the influence of the column yielding on the stress concentration ratio, settlement, and average degree of consolidation of the DM column foundation. The stress concentration ratio increased and then decreased to reach a constant value with the increase of the column modulus and time. A simplified method was proposed to calculate the maximum stress concentration ratios under undrained and drained conditions considering the column yielding. The simplified method based on a composite foundation concept could conservatively estimate the consolidation settlement. An increase of the column modulus, area replacement ratio, and/or column permeability increased the rate of consolidation.

• Food Science and Preservation
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• v.18 no.6
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• pp.954-960
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• 2011

The antioxidant activities of the ethanol, methanol, and water extracts of black onion were investigated. The highest extraction yield (28.56%) and total polyphenol content (13.5 mg/g) were found in the water extract. The water extract also showed the highest protective effect on the RAW 264.7 cell anti-inflammatory activity, and the water and ethanol extracts showed the highest reducing power. The water extract showed higher DPPH and ABTS radical scavenging activities (65.98 and 89.69%, respectively) than the other solvent extracts (ethanol, 47.77%; methanol, 66.12%). Hence, black onion can be used as a potent natural antioxidative source.

• Journal of Applied Biological Chemistry
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• v.64 no.3
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• pp.309-316
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• 2021

Kaempferol 3-O-galactoside (Trifolin), a member of the flavonol group, has been reported to have anticancer effects against promyelocytic leukemia, histocytic lymphoma, skin melanoma and lung cancer. Trifolin has been extracted and used from several plants, but the extraction process is complicated and the final yield is low. Biotransformation is an alternative tool to produce high value-added chemicals from inexpensive compounds. To synthesis trifolin from naringenin, three genes (PeFLS and OsUGE-PhUGT) were introduced into Escherichia coli, respectively. In order to synthesis trifolin from naringenin, a co-culture fermentation system was established by optimizing the cell concentration, biotransformation temperature and medium, isopropyl-β-D-thiogalactoside (IPTG) concentration, substrate supply concentration, and recombinant protein induction time. The established optimal conditions for trifolin production were a 3:1 ratio of BL-UGTE to BL-FLS, induction of recombinant protein at 25 ℃ for 4 h after addition of 2.0 mM IPTG, biotransformation at 30 ℃, and supply of 300 μM naringenin. Through the optimized co-culture fermentation system, trifolin was biosynthesized up to 67.3 mg/L.

• Journal of Microbiology and Biotechnology
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• v.28 no.12
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• pp.1999-2008
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• 2018

The compound 2,5-furandicarboxylic acid (FDCA), an important bio-based monomer for the production of various polymers, can be obtained from 5-hydroxymethylfurfural (HMF). However, efficient production of FDCA from HMF via biocatalysis has not been well studied. In this study, we report the identification of key genes that are involved in FDCA synthesis and then the engineering of Raoultella ornithinolytica BF60 for biocatalytic oxidation of HMF to FDCA using its resting cells. Specifically, previously unknown candidate genes, adhP3 and alkR, which were responsible for the reduction of HMF to the undesired product 2,5-bis(hydroxymethyl)furan (HMF alcohol), were identified by transcriptomic analysis. Combinatorial deletion of these two genes resulted in 85.7% reduction in HMF alcohol formation and 23.7% improvement in FDCA production (242.0 mM). Subsequently, an aldehyde dehydrogenase, AldH, which was responsible for the oxidation of the intermediate 5-formyl-2-furoic acid (FFA) to FDCA, was identified and characterized. Finally, FDCA production was further improved by overexpressing AldH, resulting in a 96.2% yield of 264.7 mM FDCA. Importantly, the identification of these key genes not only contributes to our understanding of the FDCA synthesis pathway in R. ornithinolytica BF60 but also allows for improved FDCA production efficiency. Moreover, this work is likely to provide a valuable reference for producing other furanic chemicals.

• Asian-Australasian Journal of Animal Sciences
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• v.32 no.3
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• pp.442-451
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• 2019

Objective: This study was conducted to investigate the effect of transport stress on physiological and hematological responses and milk performance in lactating dairy cows. Methods: Ten lactating dairy cows were randomly divided into 2 groups. The treatment group (TG) was transported 200 km for 4 h by truck, and the control group (NTG) was restrained by stanchion for 4 h in Konkuk University farm. Blood and milk samples were collected at 24 h pre-transport; 1, 2, and 4 h during transport; and 2, 24, and 48 h post-transport. Milk yields were measured at 24 h pre-transport, 0 h during transport, and 24, 48, and 72 h post-transport. Results: Leukocyte, neutrophil, and monocyte numbers in the TG were significantly higher than those of the NTG at each experimental time point. Lymphocyte numbers in the TG were significantly (p<0.05) higher than those of the NTG at 48 h post-transport. Additionally, the neutrophil:lymphocyte ratio of the TG was 45% and 46% higher than that of the NTG at 4 h during transport and 2 h post-transport, respectively. There were no significant differences in erythrocyte numbers, hemoglobin concentrations, platelet numbers, and hematocrit percentages between two groups. Cortisol levels in the TG were significantly (p<0.05) higher than those in the NTG. Milk yields in the TG were lower than those in the NTG. The somatic cell count (SCC) of the TG was significantly (p<0.05) higher than that of the NTG at 1 and 2 h during transport; that of the TG increased dramatically at 1 h during transport and gradually decreased subsequently. Conclusion: Transport stress increased blood parameters including leucocyte, neutrophil, and monocyte numbers by increased cortisol levels, but did not affect erythrocytes, hemoglobin and hematocrit levels. Additionally, transport resulted in a decrease in milk yield and reduced milk quality owing to an increase in milk SCC.