• 한국생물공학회:학술대회논문집
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• 2003.04a
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• pp.639-642
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• 2003

Olfactory receptor protein ODR10 was expressed in E.coli as fusion protein with GST and His6 Tag. Crude membrane extract of the expressed protein was coated on the surface of quartz crystal microbalance, and the interaction of the ODR10 with several odorants was examined. Although the expression level was very low, quartz crystal microbalance showed that the expressed protein interacted most strongly with diacetyl (butanedione), which is known to bind to the ODR10 protein selectively. The interaction between ODR10 and diacetyl was $5{\sim}10$ times stronger than the interaction between ODR10 and other odorants. Thus, E. coli cells expressing the olfactory receptor protein could be used as an olfactory biosensor. Also, such system could be used to test which olfactory receptor reacts specifically with which odorant molecules, since there has been no cheap and convenient way to test the interaction of olfactory receptors and odorant molecules yet.

• 한국생물공학회:학술대회논문집
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• 2003.04a
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• pp.668-672
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• 2003

An antioxidative activity of Portulaca oleracea was tested by in vitro experimental models. The antioxidative activities were determined by evaluation the DPPH radical scavenging activity and by measuring lipid peroxide using 2-thiobarbituric acid (TBA). The crude extract was sequentially partitioned with n-hexane, 15% aq. MeOH, EtOAc, n-BuOH, $H_2O$. Among them, a remarkable antioxidative effect was observed in the fractions of EtOAc and n-BuOH. The DPPH radical scavenging effect $(IC_{50}=17.90{\mu}g/ml)$ of the n-BuOH soluble fraction was comparable with that of natural antioxidant, ${\alpha}-tocopherol(IC_{50}=\;6.99{\mu}g/ml)$ and the inhibitory effect of lipid peroxidation in mouse liver homogenate was similar to that of natural antioxidant, L-ascorbic acid at a concentration of 0.1mg/ml to 5mg/ml. From the BuOH soluble fraction yielded two biophenolic glycosides, 3-hydroxy-1-(2-hydroxyethyl)phenyl-4-O-${\beta}$-D-glucopyranoside(1) and 2-(3,4-dihydroxyphenyl)ethyl-O-${\beta}$-D-glucopyranoside(2) using column chromatography and revered-phase HPLC. In particular, the DPPH radical scavenging activity of 2 was comparable to that tocopherol$(IC_{50}=6.59{\mu}g/ml)$.

• Journal of the Korean Society of Food Science and Nutrition
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• v.31 no.1
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• pp.50-56
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• 2002

The $\beta$-galactosidase was purified from the internal organs of edible snail by fractionation with ammonium sulfate, ion exchange chromatography on DEAE-Sephadex, Mono Q HR 5/5 and gel filtration on Sephacryl S-200. Superose 12 HR 10/30 chromatography. The specific activity of the purified $\beta$-galactosidase was 18.8 units/mg protein with 31.3 purification fold from crude extract. The $\beta$-galactosidase had native molecular weight of 144,000 dalton and was composed of two subunits of 72,000 dalton. The isoelectric point of the enzyme was determined 4.1. This enzyme was the most active at pH 3.0 and 6$0^{\circ}C$, and was stable in the pH range 2.0~8.0 and below 5$0^{\circ}C$. The enzyme was inhibited by metal ions and sugars such as fructose, glucose, galactose, maltose and xylose.

• Analytical Science and Technology
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• v.28 no.2
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• pp.98-105
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• 2015

The development of an enzymatic biosensor for dopamine determination based on multiwall carbon nanotubes (MWCNTs) and peroxidase obtained from the crude extract of burley tobacco (Nicotiana tabacum L.) was proposed. Peroxidase catalyzes the oxidation of dopamine to dopamine quinone. The influence on the response of analytical parameters of biosensors such as enzyme concentration, dopamine concentration, pH, and phosphate buffer solution concentration were investigated. The analytical parameters obtained, including sensitivity, linearity, and stability, were investigated. The proposed method for dopamine determination presented good selectivity even in the presence of uric acid and ascorbic acid. The sensor presented a higher response for dopamine in 0.010 M phosphate buffer at pH 6.50, with an applied potential of -0.15 V. The detection limit of the electrode was 2.7×10⁻⁶ M (S/N = 3) and the relative standard deviation of the measurements, which were repeated 10 times using 5.0×10⁻² M dopamine, was 1.3%.

• Journal of Animal Science and Technology
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• v.47 no.3
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• pp.409-418
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• 2005

This study was conducted to examine the nutrient digestibility and ruminal fermentation characteristics in sheep fed dried mugwort and mugwort silage for 5% levels of rice straw in the basal diet, and mugwort pellet for 5% levels of concentrate in the basal diet. For the experiment, they were given a basal diet containing of rice straw and concentrate mixed at a 3: 7 ratio (DM basis). The treatments were designed as a 4 ${\times}$ 4 Latin square design with four sheep (50.2 kg body weight). The digestibility of crude protein was increased (p < 0.05) to 4.6 - 6.2 % in sheep fed mugwort silage treatments (60.23 %) compared with those of control (54.08 %) and dried mugwort treatment (55.67 %). That of ether extract was iicreased (p < 0.05) to 4.8 - 8.8 % in sheep fed mugwort silage treatments (80.22 %) compared with those of control (71.47 %) and dried mugwort treatment (75.46 %). In the dry matter intake, mugwort silage treatment (904.44 g) was the hightest and mugwort pellet treatment, dried mugwort treatment and control were 810.66 g, 780.66 g and 742.18 g, respectively. The ruminal pH in all treatments were rapidly decreased (p < 0.05) at 0.5 and 1 hour after feeding and slowly increased at 2, 4 and 8 hours after feeding, especially mugwort silage treatment. The ammonia nitrogen concentrations were the highest (p < 0.05) in sheep fed mugwort silage treatment (11.24 - 12.05mg / 100 rnz) at 0.5 and 2 hours after feeding. The ruminal concentrations of acetic acid (6.06 mmol /100 $m\ell$) and propionic acid (2.35 mmol/ 100 mz) were an increased (p < 0.05) at the mugwort silage treatments at 1 and 2 hours after feeding. Purine derivatives out put (13.41 mmol / d) and microbial protein production (11.61 mmol / d) were increased (p < 0.05) compared with those of control (5.42 and 4.93 mmol / d).

• Korean Journal of Food Science and Technology
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• v.20 no.6
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• pp.762-768
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• 1988

The lipolytic enzyme of milk from hormone treated and non treated cows was isolated and purified, It was shown that the crude lipase extract from the milk before and after a hormone treatment of the cows was different in color, foaming properties, yield and specific activity. Final purification of the lipase system was achieved by affinity chromatography on Heparin-Sepharose CL-6B. The lipase bound by Heparin-Sepharose was then characterised. The pH-optimum of the purified enzyme was 8.5 for butteroil emulsion as a substrate and the optimum temperature was $30^{\circ}C$ respectively. The molecular weight. determined by SDS-polyacrylamidegel electrophoresis, was about 70,000. The activity increased by 10% hen 0.01% bovine serum albumin was added to the substrate. The results indicate the enzymes obtained by affinity chromatography from milk before and after hormone treatment had the similar characteristics. The second lipolytic active component that was not bound by Heparin-Sepharose must be the cause of spontaneous rancidity.

• Journal of Pharmacopuncture
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• v.22 no.4
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• pp.253-259
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• 2019

Objectives: It is reported that tumor-associated macrophages (TAMs) contribute to cancer progression by promoting tumor growth and metastasis. The purpose of this study is to investigate the effect of different fractions of Adenophora triphylla var. japonica (AT) on the polarization of macrophages into the M2 phenotype, a major phenotype of TAMs. Methods: We isolated hexane, ethyl acetate, and butanol fractions from crude ethanol extract of AT. The cytotoxicity of AT in RAW264.7 cells was examined by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. RAW264.7 cells were polarized into the M2 phenotype by treatment with interleukin (IL)-4 and IL-13. The expression of M2 macrophage marker genes was detected by reverse transcription polymerase chain reaction (RT-PCR). The phosphorylation level of signal transducer and activator of transcription 6 (STAT6) was investigated by western blot analysis. The migration of Lewis lung carcinoma (LLC) cells was examined by transwell migration assay using conditioned media (CM) collected from RAW264.7 cells as a chemoattractant. Results: Among various fractions of AT, the ethyl acetate fraction of AT (EAT) showed the most significant suppressive effect on the mRNA expression of M2 macrophage markers, including arginase-1, interleukin (IL)-10 and mannose receptor C type 1 (MRC-1), up-regulated by treatment of IL-4 and IL-13. In addition, EAT suppressed the phosphorylation of STAT6, a critical regulator of IL-4 and IL-13-induced M2 macrophage polarization. Finally, the increased migration of Lewis lung carcinoma (LLC) cells by CM from M2-polarized RAW264.7 cells was reduced by CM from RAW264.7 cells co-treated with EAT and M2 polarization inducers. Conclusion: We demonstrated that EAT attenuated cancer cell migration through suppression of macrophage polarization toward the M2 phenotype. Additional preclinical or clinical researches are needed to evaluate its regulatory effects on macrophage polarization and anti-cancer activities.

• Parasites, Hosts and Diseases
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• v.38 no.2
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• pp.75-84
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• 2000

Antibody responses in serum and cerebrospinal fluid (CSF) samples from patients with active and chronic paragonimiasis and in sera from patients on whom follow-up studies were done after praziquantel treatment were analyzed using antigens of Paragonimus westermani prepared from eggs, metacercariae, juveniles of 4-and 7-week old, adult worms and recombinant protein of 28 kDa cruzipain-like cysteine protease (rPw28CCP). The patient sera/CSFs of active and chronic paragonimiasis revealed strong antibody reactions against the crude extracts of 4-and 7-week old juveniles as well as against those from egg and adult. rPw28CCP also showed specific reaction to the sera with active paragonimiasis. After the treatment, levels of specific antibodies in the sera gradually decreased to negative range in most patients. In some cases with persisting high antibody levels, however, the reactions at 27 kDa egg Protein were sustained throughout the observation period of 34 months. The reactions at 35 and 32 kDa in adult extract and rPw28CCP disappeared rapidly after the treatment. Persistent antibody reactions even after successful treatment are provoked by continuous antigenic challenge from eggs which were not resolved by treatment.

• Korean Journal of Medicinal Crop Science
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• v.19 no.6
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• pp.484-490
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• 2011

The potential antioxidant activities of different fractions from Prunella vulgaris var. lilacina were assayed in vitro. Among several fractions, n-BuOH fraction showed the highest 1,1-di[henyl-2-picrylhydrazyl (DPPH) free radical scavenging ($IC_{50}=0.50{\mu}g/mL$). The results of 2,2'-azinobis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) radical scavenging activity and ferric reducing antioxidant power (FRAP) assay showed the concentration dependency and n-BuOH fraction appeared a better result than the other fractions at the same concentrati on in this study. Moreover the total phenol and flavonoid contents of n-BuOH fraction contained the highest level. Additionally, correlation analysis indicated a high correlation between the antiradical activity and the total phenolic and flavonoid contents (p < 0.001). It suggests that n-BuOH fraction obtained from the 70% EtOH crude extract of Prunella vulgaris var. lilacina has wide potential for use as a source of antioxidant material.

• Asian-Australasian Journal of Animal Sciences
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• v.23 no.7
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• pp.889-900
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• 2010

Spices can be used as novel supplements to enhance the utilization of low quality forages (LQF) and reduce nutrient wastage by ruminant animals. However, it is essential to characterize these spices alongside LQF before testing their potential use as supplements in ruminant diets. This study characterized four spices (cinnamon, cumin, clove and turmeric) alongside three forages (rice straw, wheat straw and hay) for their chemical components before evaluating their effect at four different doses (0, 10, 30 and 90 mg/g forage DM) on the in vitro rumen degradability of dry matter (DM) (IVD) and organic matter (OM) (IVOMD) of these forages at various incubation times. It appeared that some spices could provide complementary nutrients which could improve the utilization of LQF where hay had better chemical composition than the other two forages. Cumin contained more crude protein (CP), ether extract and mineral contents whereas turmeric contained more soluble sugars than the other spices. Cinnamon was least acceptable as a ruminant supplement due to its higher condensed tannin and saponin and lower CP and mineral contents. The IVD and IVOMD were highest for hay and lowest for wheat straw with all spices at all incubation times (p<0.001). Due to relatively better nutrient profiles, cumin and turmeric had greater effect on IVD and IVOMD of the forages. In contrast, cinnamon had negative effects on IVD and IVOMD. IVD and IVOMD were greater at 10 mg/g than at other levels of most spices suggesting that using certain amounts of spices can increase forage degradability. However, the choice of a spice will depend upon the forage type being offered to ruminants. Further studies will examine the effect of these spices on fermentation profile, methane production and nitrogenous loss by ruminants.