• Bulletin of the Korean Chemical Society
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• v.29 no.8
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• pp.1510-1518
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• 2008

Results of intrinsic and extrinsic fluorescence studies on choline oxidase revealed that the enzyme at high alkaline pH values has more accessible hydrophobic patches relative to acidic pH. Fluorescence quenching studies with acrylamide confirm these changes. The quenching constants were also determined at different pH(s) by using the Stern-Volmer equation. CD studies showed that at higher pH a transition from $\alpha$-helix to $\beta$- structure was appeared while at lower pH the content of $\alpha$-helix structure was increased. Furthermore, analysis of the spectral data using chemometric method gave evidence for existence of intermediate components at very high pH(s). Results of thermal denaturation evaluated that the enzyme has the most instability at higher pH(s). Altogether low and high pH values caused significant alteration on secondary and tertiary structures of choline oxidase via inducing of an intermediate.

• Journal of Nutrition and Health
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• v.41 no.5
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• pp.428-438
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• 2008

Choline is important for normal membrane function, acetylcholine synthesis and methyl group metabolism. In this study, 185 food items customarily eaten by Koreans were selected from the data of the 2001 Korean National Health and Nutrition Survey and analyzed on the total choline content of the foods using enzymatic method of choline oxidase. Foods with high choline concentration (mg/100 g) were listed in sequence of quail egg (476.04 mg), dried squid (452.42 mg), beef liver (427.16 mg), pork liver (424.92 mg), tuna canned in oil (414.44 mg), boiled and dried anchovy (381.30 mg), dried Alaskan pollack (378.88 mg), chicken egg (309.88 mg), chicken liver (259.38 mg), soybean (238.62 mg), French bread with garlic (193.18 mg) and barley (183.73 mg). From this result, it is shown that dried fishes, prepared fishes, livers, eggs, pulses and cereals might be categorized as high choline food. Citron tea and green tea showed low choline content below 1 mg. Vegetables and fruits were also categorized into low choline food. No choline was detected in red pepper powder, beer, soju, soybean oil and corn oil out of foods analyzed in this study. Further study is required for analytic procedure of the foods of which results are inconsistent with USDA's data such as rice and wheat flour.

• YAKHAK HOEJI
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• v.56 no.4
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• pp.222-229
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• 2012

In order to determine acetylcholine and choline in the biological samples, the specific enzymes of acetylcholinesterase (AChE) and choline oxidase (ChO), which utilize acetylcholine and choline as substrates, were employed to convert substrates to $H_2O_2$. The produced $H_2O_2$ was coupled to 4-aminoantipyrine/phenol with peroxidase (PO) yielding quinoneimine dye which was measured at 508 nm. In the present enzymatic spectrophotometric analysis the product at the equilibrium state was measured considering accuracy, precision, time and cost of the analysis. The developed analytical method yielded good linearity (calibration curve; $A_{508}$=9534[acetylcholine]+0.009, correlation coefficient ($R^2$); 0.999) with detection limit of $1.11{\times}10^{-7}M$, reasonable precision (relative standard deviation; 0.10~1.62% at $2.5{\times}10^{-6}M{\sim}1.25{\times}10^{-4}M$) and accuracy (relative error; -0.24~0.97% at $4.13{\times}10^{-6}M{\sim}1.01{\times}10^{-4}M$) for acetylcholine chloride standard solution. The concentrations of acetylcholine and choline in human serum were found as $3.20{\times}10^{-5}M$ and $1.14{\times}10^{-4}M$, respectively. The brain tissues of Sprague-Dawley strain rat contained 9.82${\mu}g/g$ of acetylcholine and 6.53 ${\mu}g/g$ of choline in the cerebrum, while 7.37 ${\mu}g/g$ of acetylcholine and 5.34 ${\mu}g/g$ of choline in the cerebellum.

• Proceedings of the Korean Society of Plant Biotechnology Conference
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• 2005.11a
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• pp.187-187
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• 2005

• Journal of Nutrition and Health
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• v.37 no.2
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• pp.95-99
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• 2004

This study was designed to investigate the effects of ethyl acetate(EtOAc) fraction of pine (Pinus densiflora Sieb et Zucc) needle on cholesterol and lipofuscin(LF) accumulations, acetylcholine(ACh) and its related enzyme activities such as choline acetyltransferase(ChAT) and acetylcholinesterase(AChE), and monoamine oxidase-B(MAO-B) activity, which destroyed the catecholamine related neurotransmitters in brain membranes of Sprague- Dawley (SD) rats. Male SD rats were fed basic diets (control group) and experimental diets (EtOAc-25, EtOAc-50 and EtOAc-100) for 45 days. Cholesterol accumulations in mitocholndria and microsomes were significantly inhibited (11.8-12.1% and 9.6-13.0%, respectvely) in EtOAc-50 and EtOAc-100 groups. ACh levels and ChAT activities were significantly increased about 10% in membranes of EtOAc-100 group compared with control group. AChE activities were significantly increased about 8 -12% in membranes of EtOAc-50 and EtOAc-100 groups compared with control group. MAO-B activities were significantly inhibited about 10% in membrane of EtOAc-l00 group compared with control group. These results suggest that ethyl acetate fraction of pine needle may play an effective role in inhibiting cholesterol and improving a membrane fluidity, and learning and memory impairments. (Korean J Nutrition 37(2): 95 -99, 2004)

• Journal of Plant Biotechnology
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• v.42 no.1
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• pp.19-24
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• 2015

Glycine betaine (GB) is one of the compatible solutes that accumulate in the chloroplasts of certain halotolerant plants under salt or cold stress. The codA gene for choline oxidase, the enzyme that converts choline into GB, has been cloned from a soil bacterium Arthrobacter globiformis. We generated transgenic sweetpotato plants [Ipomoea batatas (L.) Lam] expressing codA gene in chloroplasts under the control of the SWPA2 promoter (referred to as SC plants) and evaluated SC plants under oxidative and drought stresses. SC plants showed enhanced tolerance to methyl viologen (MV)-mediated oxidative stress and drought stress due to induced expression of codA. At $5{\mu}M$ of MV treatment, all SC plants showed enhanced tolerance to MV-mediated oxidative stress through maintaining low ion leakage and increased GB levels compared to wild type plants. When plants were subjected to drought conditions, SC plants showed enhanced tolerance to drought stress through maintaining high relative water contents and increased codA expression compared to wild type plants. These results suggest that the SC plants generated in this study will be useful for enhanced biomass production on global marginal lands.

• Korean Journal of Food Science and Technology
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• v.21 no.5
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• pp.726-734
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• 1989

To analyze the components of dense coat fractions associated with fat globule membrane, The membrane was treated with various concentrations of Triton X-100, a non-ionic detergent, and the composition of lipids associated to the detergent insoluble material was analyzed. The amount of protein, phospholipid, cholesterol and ganglioside in milk fat globule membrane was reduced consistently with increasing concentrations of Triton X-100. Butyrophilin (band 12), xanthine oxidase (band 3) and band 16 as constituents of insoluble coat materials was revealed after electrophorisis on SDS-polyacrylamide gels. Phosphatidyl ethanolamine, phosphatidyl choline, phosphatidyl serine, phosphatidyl inositol and sphingomyelin were identified as the major phospholipids of the coat materials without selective concentration relative to the original membranes. Percentages of total phospholipid were not changed by any of the treatments. Fatty acids of total lipid were myristate, palmitate, stearate (major saturated acids), oleate and linoleate (major unsaturated acids). Cholesterol contents on a protein basis were slightly reduced with increasing concentrations of Triton X-100. Cholesterol adhered to protein more tightly than other constituents The contents of gangliosides was proportionally refuted with increasing concentration of Triton X-100.

• Journal of Life Science
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• v.10 no.6
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• pp.564-569
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• 2000

This study was designed to investigate the effect of silkworm (Bombyx mori L.) powder on lipofuscin, acetylcholine (ACh) and its related enzyme activities in brain of rats. Sprague-Dawley (SD) male rats ($160{\pm}10g$) were fed basic diet (control group), and experimental diets (SWP-200 and SWP-400 groups) added 200 and 400 mg/kg BW/day for 6 weeks. In case of liver membranes, lipofuscin (LF) levels resulted in a slight decreases (4.6% and 11.5%, respectively) in SWP-200 and SWP-400 groups compared with control group. But in case of barin as the most sensitive organ, LF levels were remarkably inhibited about 16.7% and 20.0% in SWP-200 and SWP-400 groups compared with control group. There were no significant differences in acetylcholine (ACh) syntheses as a very important neurotransmitter, and choline acetyltransferase (ChAT) activities as a synthesis enzyme of ACh, and acetylcholinesterase (AChE) activities as a hydrolysis enzyme, which were concerned in transmission of neuron through synapses in brain of SWP-200 and SWP-400 groups compared with control group. Monoamine oxidase-B (MAO-B) activities were significantly inhibited (about 10.2%) in brain of SWP-400 groups compared with control group. These results suggest that inhibiting effects of LF accumulation and MAO-B activity of silkworm powder (SWP) may play a pivotal role in attenuating a various age-related changes for improvement of brain function.

• Journal of Nutrition and Health
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• v.37 no.3
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• pp.176-181
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• 2004

This study was designed to investigate the effects of buthanol (BuOH) fraction of pine (Pinus densiflora Sieb et Zucc) needle on cholesterol and lipofuscin (LF) accumulations, acetylcholine (ACh) and its related enzyme activities such as choline acetyltransferase (CAhT) and acetylcholinesterase (AChE), and monoamone oxidase-B (MAO-B) activity, which destroyed the catecholamine-related neurotransmitters in brain membranes of Sprague-Dawley (SD) rats. Male SD rats were fed basic diets (control group) or experimental diets (BuOH-25, BuOH-50 and BuOH-100) for 45 days. Cholesterol accumulations in mitochondria and microsomes were significantly inhibited (about 14 - 17％ and 23 - 34％, respectvely) in BuOH-50 and BuOH-100 groups, whereas LF levels were significantly inhibited (about 10 - 14％) in BuOH-50 and BuOH-100 groups compared with control group. ACh levels and ChAT activities were significantly increased (about 11 - 17％ and 11 - 23％, respectively) in membranes of BuOH-50 and BuOH-100 groups compared with control group. AChE activities were significantly increased (about 14 - 17％) in membranes of BuOH-50 and BuOH-100 groups. There was no significant difference in MAO-B activities between control and experimental diet groups. The results suggest that butanol fraction of pine needle may play an effective role in an antiaging effect and improving a learning and memory impairments.

• Journal of Sericultural and Entomological Science
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• v.42 no.2
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• pp.120-125
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• 2000

This study was designed to investigate the effects of silk fibroin (Mw 500) powder (SFP) on lipofuscin, acetylcholine (ACh) and its related enzyme activities in brain of rats. Sprague-Dawley (SD) male rats (160$\pm$10 g) were fed basic diet (control group), and experimental diets (SFP-2.5 and SFp-5.0 groups) added 2.5 and 5.0 g/kg BW/day for 6 weeks. In case of liver membranes, lipofuscin (LF) levels resulted in a considerable decreases (11.5% and 13.8%, respectively) in SFP-2.5 and SFP-5.0 groups compared with control group. But in case of brain as the most sensitive organ, LF levels were remarkably inhibited about 18.3% and 21.7% in SFP-2.5 and SFP-5.0 groups compared with control group. Acetylcholine (ACh) levels were considerable decrease (3.0% and 9.2%, respectively) in brain membranes of SFP-2.5 and SFP-5.0 groups compared with control group. choine acetyltranferase (ChAT) activities as a synthesis enzyme of ACh, and acetylcholinesterase (AChE) activities as a hydrolysis enzyme resulted in a slight increases (2.4% and 3.0%, 4.6% and 6.3%, respectively), but significance difference between ChAT and AChE activities by SFP administration could be not obtained. Monoamine oxidase-B (MAO-B) activities were significantly inhibited (9.5% and 12.6%, respectively) in brain of SEP-2.5 and SFP-5.0 groups compared with control group. These results suggest that inhibiting effects of LF accumulation and MAO-B activity of silk fibroin(SFP) may play a pivotal role in protecting learning memory impairments by attenuating a various age-related changes for improvement of brain function.