• Applied Biological Chemistry
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• v.26 no.1
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• pp.19-27
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• 1983

The lipid and fatty acid compositions of neutral lipid, glycolipid and phospholipid were identified and quantified by thin-layer chromatography and gas chromatography. Main constituents of the neutral lipid were triglyceride, free sterol and esterified sterol in the free lipid, and free fatty acid and monoglyceride in the bound lipid. In the outer part, there existed 25.7% triglyceride in the free lipid and it was not nearly found in the bound lipid. Main constitutents of the glycolipid were digalactosyl diglyceride and esterified steryl glycoside in the free lipid, and digalactosyl diglyceride in the bound lipid. Free lipid didn't contain trigalactosyl diglyceride but bound lipid contained 2.0% of it. Main constituents of the phospholipid were lysophosphatidyl choline, phosphatidyl inositol, phosphatidyl choline, phosphatidyl glycerol and phosphatidyl ethanolamine in the free lipid, and phosphatidyl choline and phosophatidyl inositol in the bound lipid. The predominant fatty acids of three fractions, neutral lipid, glycolipid and phospholipid showed almost the same pattern as that of the total free and bound lipids. The content of palmitic acid was relatively higher in the polar lipids(glyco and phospholipid). Therefore, saturated fatty acid ratio of polar lipid was higher than that of neutral lipid. found lipid contained more saturated fatty acids as compared with the free lipid.

• Animal cells and systems
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• v.2 no.2
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• pp.233-238
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• 1998

Certain basic characteristics of choline uptake in nerve terminals were studied with synaptosomes from rat hippocampus. Synaptosomal $[^3H]$-choline uptake was clarified as specific and high affinity by low Km value(2.2 uM), Na+-dependency and high sensitivity to hemicholinium-3, a competitive inhibitor of choline uptake. Choline uptake into synaptosomes was linearlys related to Na+ concentration and membrane potential. Extracellular Ca2+ modulated the choline uptake, but probably not through increase of intracellular $Ca^{2+}$, because this modulation was not affected the by high $K^+$-depolarization. EGTA (2mM) added for $Ca^{2+}$-free condition had a peculiar effect of decreasing choline uptake. These results suggest that Ca2+ may play an important role in regulating the metabolism of acetylcholine in the nerve terminals directly through the increase of acetylcholine release.

• Journal of Plant Biology
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• v.29 no.3
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• pp.157-165
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• 1986

The onset of photophosphorylation at the various stages of greening showed different patterns with varying concentrations of Pi. With further greening, ATP formation occurred at the lower concentration of Pi (48 hrs; 0.05 mM). At early stages of greening, more Pi was required for photophosphorylation (6 hrs; 5.0 mM). The addition of cell-free extracts of etiolated barley seedlings resulted in the competitive inhibition of photophophorylation. The apparent inhibition by cell-free extracts were gradually decreased during greening of etiolated barley seedlings. We found that the inhibitors of photophosphorylation in cell-free extracts were some organic phosphates and most of them was P-choline. P-choline inhibited photophosphorylation competitively with Pi and its content was decreased considerably in greening. It is likely that P-choline partly delay the photophorylation in early stages of greening.

• Journal of the Korean Applied Science and Technology
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• v.3 no.2
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• pp.49-64
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• 1986

This experiment is carried out to study effect of choline-deficient diet on serum and liver lipid contents of male rats. The experimental animals use 36 male Sprague-Dawley rats weighing about $100{\pm}3g$. They are classified into 7 groups and fed to experimental diets which are composed of 0.8% choline-supplement of deficient diets in addition to 14% corn oil, 14% corn margarine and 14% lipids mixed with 4% corn oil and 10% corn margarine, respectively. After feeding for 4 weeks, I measure lipid concentration of serum and liver, and the result are as follows. 1. The choline-deficient diet group decreases slightly the rates of weight gain and feed efficiency as compared with those of the choline-supplement diet group, but increases liver weight. 2. The choline-dificient diet group decreases the serum concentrations of total cholesterol (TC), free cholesterol, HDL-choelsterol, VLDL, LDL-cholesterol and phospholipid (PL), but increases the contents of triglyceride and the ratios of cholesterol/HDL-cholesterol and triglyceride/PL, and indicates no remarkable-difference in the ratio of TC/PL. 3. As compared with the choline-supplement diet group, the choline-deficient diet group contains the higher liver contents of total lipids, free cholesterol and triglyceride, and gives little difference in the liver contents of total cholesterol and phospholipid(PL), and presents the higher ratios of VLDL, LDL-cholesterol/HDL-cholesterol and TG/PL in the liver.4. In the choline-deficient diet group, the coutents of serum and liver lipid is not influenced by the kind of dietary fat. On the other hand, the choline-supplemented diet group indicates a significantly lower content of phospholipid in the corn margarine-added diet group than in the corn oil-added diet group. As aforementioned results, I think that the choline-deficient diet induces fatty liver in male rats without relation to kind of fatty acid, and cholines-upplement diet with saturated fatty acid makes the more decrease of liver phospholipid than that with polyunsaturated fatty acid.

• Journal of Nutrition and Health
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• v.23 no.6
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• pp.418-426
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• 1990

This study examines the effects of choline deficiency and 2-acetylaminofluorene(2-AAF) on the lipid peroxide values, glucose 6-phosphatase(G6Pase) and glutathione S-transferase (GST) activities in rats fed different dietary fats. Weanling Sprague Dawley male rats fed the diets containing 15% beef tallow or 15% corn oil with vitamin fortification mixture or choline free vitamin mixture for 10 weeks. At 3th and 5th week, 2-AAF was injected twice each week intraperitoneally. Total 2-AAF injection was four times. 2-AAF and choline deficiency increased lipid peroxidation in corn oil groups, so the role of 2-AAF and choline deficiency in lipid peroxidation was more important in corn oil groups than beef tallow groups. G6Pase activities tended to be decreased by 2-AAF in choline deficient groups, and in corn oil groups, the enzyme activities were decreased significantly in all subgroups compaired to beef tallow groups. GST activities were increased by 2-AAF in beef tallow groups and choline deficiency in corn oil groups, and might defence against carcinogen metabolism and lipid peroxidation.

• Journal of Nutrition and Health
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• v.43 no.6
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• pp.588-596
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• 2010

Most nutrients taken by pregnant women are secreted into their breast milk. Food contains choline together with betaine, and in human body choline is oxidized to betaine which transfer methyl group. The aim of the study was to estimate the concentrations of choline and betaine in breast milk of Korean lactating women and the choline and betaine intakes of their infants. Total choline, free choline and betaine concentrations in breast milk of some lactating women living in Daejon Metropolitan city were analyzed every month by using HPLC-MS and enzymatic method during the first five months. Total choline concentrations of breast milks were 157.64 mg/L (1.52 mmol/L), 157.83 mg/L (1.52 mmol/L), 165.99 mg/L (1.60 mmol/L), 153.67 mg/L (1.48 mmol/L), 145.05 mg/L (1.39 mmol/L) by month after delivery for five months. The concentrations of total choline and free choline in breast milks were not significantly changed for the five months while the betaine concentrations gradually decreased. Daily intake of total choline of the infants appears to be adequate for the infant's requirement according to the US DRI; 124.6 mg/d, 120.9 mg/d, 126.5 mg/d 104.1 mg/d from 2nd to 5th month after birth. Free choline and betaine intakes of the infants were not significantly changed during the four months except showing decrease in betaine intake per kg body weight. Choline intakes of the infants more correlated with choline concentrations of the breast milks (r = 0.982, p = 0.000) than intake amount of the breast milk (r = 0.414, p = 0.028). These results suggest that the choline intake of Korean breast-fed infants appears to be adequate and the intake could be affected by the choline concentration of the breast milk.

• YAKHAK HOEJI
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• v.47 no.4
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• pp.224-229
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• 2003

We have investigated the transport characteristics of synthetic antioxidant and free radical scavenger, $\alpha$-phenyl-n-tert-butyl nitrone (PBN) at the blood-brain barrier (BBB) by in vitro uptake study in conditionally immortalized rat brain capillary endothelial cell line (TR-BBB). Also, the efflux of PBN from brain to blood is estimated using the brain efflux index (BEI) method. Choline is a charged organic cation, including nitrogen-methyl group and shows the carrier-mediated distribution to the brain. [$^3$H]Choline uptake by TR-BBB cells was significantly inhibited by PBN with $IC_{50}$/ of 1.2 mM, which appears to be due to similar structures between choline and PBN. And, PBN was microinjected into Par2 of the rat brain by BEI method, and was eliminated from the brain with an apparent elimination half-life of about 2 min. Also, [$^3$H]choline efflux was significantly inhibited by PBN using BEI method. In conclusion, the efflux transport of PBN takes place across the BBB and PBN may be transported into the brain and eliminated from the brain by BBB choline transporter.

• Asian-Australasian Journal of Animal Sciences
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• v.31 no.5
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• pp.686-695
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• 2018

Objective: The objective of this study was to investigate the effects of dietary choline supplementation on hepatic lipid metabolism and gene expression in finishing pigs with intrauterine growth retardation (IUGR). Methods: Using a $2{\times}2$ factorial design, eight normal birth weight (NBW) and eight IUGR weaned pigs were fed either a basal diet (NBW pigs fed a basal diet, NC; IUGR pigs fed a basal diet, IC) or a diet supplemented with two times more choline than the basal diet (NBW pigs fed a high-choline diet, NH; IUGR pigs fed a high-choline diet, IH) until 200 d of age. Results: The results showed that the IUGR pigs had reduced body weight compared with the NBW pigs (p<0.05 from birth to d 120; p = 0.07 from d 120 to 200). Increased (p<0.05) free fatty acid (FFA) and triglyceride levels were observed in the IUGR pigs compared with the NBW pigs. Choline supplementation decreased (p<0.05) the levels of FFAs and triglycerides in the serum of the pigs. The activities of malate dehydrogenase and glucose 6-phosphate dehydrogenase were both increased (p<0.05) in the livers of the IUGR pigs. Choline supplementation decreased (p<0.05) malate dehydrogenase activity in the liver of the pigs. Gene expression of fatty acid synthase (FAS) was higher (p<0.05) in the IC group than in the other groups, and choline supplementation decreased (p<0.05) FAS and acetyl-CoA carboxylase ${\alpha}$ expression in the livers of the IUGR pigs. The expression of carnitine palmitoyl transferase 1A (CPT1A) was lower (p<0.05) in the IC group than in the other groups, and choline supplementation increased (p<0.05) the expression of CPT1A in the liver of the IUGR pigs and decreased (p<0.01) the expression of hormone-sensitive lipase in both types of pigs. The gene expression of phosphatidylethanolamine N-methyltransferase (PEMT) was higher (p<0.05) in the IC group than in the other groups, and choline supplementation significantly reduced (p<0.05) PEMT expression in the liver of the IUGR pigs. Conclusion: In conclusion, the lipid metabolism was abnormal in IUGR pigs, but the IUGR pigs consuming twice the normal level of choline had improved circulating lipid parameters, which could be related to the decreased activity of nicotinamide adenine dinucleotide phosphate-generating enzymes or the altered expressions of lipid metabolism-related genes.

• Animal Bioscience
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• v.35 no.11
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• pp.1787-1799
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• 2022

Objective: Choline deficiency, one main trigger for nonalcoholic fatty liver disease (NAFLD), is closely related to lipid metabolism disorder. Previous study in a choline-deficient model has largely focused on gene expression rather than gene structure, especially sparse are studies regarding to alternative splicing (AS). In modern life science research, primary hepatocytes culture technology facilitates such studies, which can accurately imitate liver activity in vitro and show unique superiority. Whereas limitations to traditional hepatocytes culture technology exist in terms of efficiency and operability. This study pursued an optimization culture method for duck primary hepatocytes to explore AS in choline-deficient model. Methods: We performed an optimization culture method for duck primary hepatocytes with multi-step digestion procedure from Pekin duck embryos. Subsequently a NAFLD model was constructed with choline-free medium. RNA-seq and further analysis by rMATS were performed to identify AS events alterations in choline-deficency duck primary hepatocytes. Results: The results showed E13 (embryonic day 13) to E15 is suitable to obtain hepatocytes, and the viability reached over 95% by trypan blue exclusion assay. Primary hepatocyte retained their biological function as well identified by Periodic Acid-Schiff staining method and Glucose-6-phosphate dehydrogenase activity assay, respectively. Meanwhile, genes of alb and afp and specific protein of albumin were detected to verify cultured hepatocytes. Immunofluorescence was used to evaluate purity of hepatocytes, presenting up to 90%. On this base, choline-deficient model was constructed and displayed significantly increase of intracellular triglyceride and cholesterol as reported previously. Intriguingly, our data suggested that AS events in choline-deficient model were implicated in pivotal biological processes as an aberrant transcriptional regulator, of which 16 genes were involved in lipid metabolism and highly enriched in glycerophospholipid metabolism. Conclusion: An effective and rapid protocol for obtaining duck primary hepatocytes was established, by which our findings manifested choline deficiency could induce the accumulation of lipid and result in aberrant AS events in hepatocytes, providing a novel insight into various AS in the metabolism role of choline.

• YAKHAK HOEJI
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• v.39 no.3
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• pp.231-242
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• 1995

Electroconvulsive shock (ECS) increases the activity of acetylchohnesterase and decreases in brain acetylcholine levels. A large amount of free fatty acids accumulated in the brain tissue affects cerebral blood flow, brain edema and inflammation and results in brain injury. The present study examined the effect of docosahexaenoic acid (DHA) and D,L-pyroglutamic acid (D,L-PCA) on the learning and memory deficit using the passive avoidance failure technique and on the change of acetylcholine and choline level in the cerebral cortex of ECS-induced mice. The application of ECS (25mA, 0.5sec) induced a significant decrease in memory function for 30 min. ECS-induced a significant decrease in cortical acetylcholine and choline levels 1 min following the ECS application, which were almost recovered to ECS control level after 30 min. DHA (20 mg/kg, i.p.). administered 24 hr before shock. prevented the ECS-induced passive avoidance failure and the decrease of acetylcholine level 1 min following the ECS application. DHA failed to elicit a change in cortical choline level. DHA did not affect memory function and the cortical Ach and choline level of normal mice. The administration of D,L-PCA (500 mg/kg, i.p.) increased the effect of DHA on memory function and the change of cortical acetylcholine level of ECS induced mice. These results suggest that DHA treatment may be contributed to the prevention against memory deficit, and to the activation of cholinergic system in the ECS induced mice.