• Korean Journal of Food Science and Technology
• /
• v.29 no.1
• /
• pp.26-31
• /
• 1997

Bee venom (Apis mellifera) phospholipase $A_2$ solubilized in reversed micelles containing small amount of water stabilized by surfactant could catalyze the hydrolysis of dipalmitoyl phosphatidylcholine (DPPC). A sensitive and simple high performance liquid chromatographic (HPLC) methodology of phospholipase $A_2$ assay for the hydrolysis of DPPC was developed. Kinetic analysis of the phospholipase $A_2$-catalyzed reaction was found to be possible in reversed micelles. Among the surfactants and organic solvents tested, aerosol-OT and isooctane were most effective for the hydrolysis of DPPC in reversed micelles. Optimal temperature, optimal pH, $K_{m,app.},\;V_{max.,app.}$ and activation energy were determined to be $35{\sim}40^{\circ}C$, 7.0, 8.73 mM, 2.83 units/㎎ protein and 12.31 kcal/mole, respectively. The hydrolysis activity was dependent on water content and maximum activity was obtained at R value (=[water]/[aerosol-OT]) of 10.0.

• Journal of Pharmaceutical Investigation
• /
• v.35 no.5
• /
• pp.369-373
• /
• 2005

The objectives of the current work is to understand the factors impacting the formulation and performance of a Carbopol mucoadhesive buccal delvery system for a model peptide drug, $[D-Ala{^2},\;D-Leu{^5}]$enkephalin (DADLE, Mw=569.7) with comparable chemical and enzymatic stability. Specifically, in vitro buccal DADLE delivery from the cross-linked poly(acrylic acid) (PAA) hydrogel system was characterized. In addition, the influences of several penetration enhancers on the ex vivo buccal absorption of DADLE were also studied. In this study, the PAA hydrogels generally swell to 100% of their original weight in the phosphate pH 7.4 buffer. The water penetration into the PAA hydrogel occurred based on a zero-order kinetics for the first 60 min and steadily decreased afterwards. From the release study, it can be seen that the initial DADLE release was so rapid and the rate of release of DADLE decreased as the time elapsed. The porcine buccal tissue was found to be permeable to DADLE with a flux value of $0.07%/cm{^2}/hr({\pm}0.01\;SD)$. From the ex vivo diffusion study, it was found that sodium taurodihydrofusidate showed a greater degree of enhancement compared to the phospholipids with an Enhancement Ratio (ER) of 8.7 compared to 2.7 and 1.9 for didecanoylphosphatidylcholine and lysophosphatidylcholine, respectively. The work encompassed within this paper has demonstrated the feasibility of using the PAA hydrogel delivery system with its good mucoadhesive properties for the buccal delivery of peptides.

• 대한방사선방어학회:학술대회논문집
• /
• 2011.11a
• /
• pp.212-213
• /
• 2011

The biological effects of low dose ionizing radiation (LDIR) remain insufficiently understood. We examined for the scientific evidence to show the biological effects of LDIR using radiation-sensitive immune cells. We found that Ikaros protein was responsed to low dose-dependent effects of gamma radiation in IM-9 B lymphocytes. Ikaros encodes zinc finger transcription factors that is important regulators of a hematopoietic stem cells (HSCs) progression to the B lymphoid lineage development, differentiation and proliferation. In this study, we observed that cell proliferation was enhanced from 10% to 20% by LDIR (0.05 Gy) in IM-9 B lymphocytes. The Ikaros protein was phosphorylated in its serine/threonine (S/T) region and decreased its DNA binding activity in the cells exposed to LDIR. We found that Ikaros phosphorylation was up-regulated by CK2/AKT pathway and the residues of ser-304 and ser-306 in Ikaros was phosphorylated by LDIR. We also observed that Ikaros protein was localized from the nucleus to the cytoplasm after LDIR and bound with Autotaxin (ENPP2, ATX) protein, stimulating proliferation, migration and survival of immune cells. In addition, we found that the lysoPLD activity of ATX was dependent on Ikaros-ATX binding activity. These results indicate that the Ikaros is an important regulator of immune activation. Therefore, we suggest that low dose ionizing radiation can be considered as a beneficial effects, stimulating the activation of immune cells.

• YAKHAK HOEJI
• /
• v.47 no.6
• /
• pp.410-416
• /
• 2003

Autotaxin(ATX) was originally purified from conditioned media of A2058 human melanoma cells and shown to be a potent cell motility-stimulating factor, possessing a type II nucleotide pyrophosphatase/phosphodiesterase (NPP2) activity. Recombinant ATX has recently demonstrated that human plasma lysophosholipase D is identical to ATX and uses lysophosphatidylcholine as a substrate to mediate various biological functions including tumor cell growth and motility through G-protein coupled receptor. However, despite pivotal roles of ATX on physiological or pathophysiological states, the production of ATX is solely depends on complicated purification method which employs multiple column steps, but resulted in very poor yield. This limited the use of ATX for extensive analysis. We, therefore, expressed six histidine-tagged recombinant human ATX(His-ATX) in High Five TM insect cells to improve the generation of ATX and to make simple the purification of ATX. The signal sequence of the human ATX gene was truncated and replaced with sequence of insect cell secretion signal within expression vector. In addition, codons for six histidines were added to the C-termini of 120kDa ATX cDNA construct. A simple purification scheme utilizing two-step affinity column chromatography was designed to purify His-ATX to homogeneity from the culture supernatant of transfected insect cells. Homogenous His-ATX was detected and isolated from the concentrated insect cell medium using concanavalin A agarose and nickel affinity chromatography. Purified His-ATX was in full length with ATX capacity. A combination of this expression system and purification scheme would be useful for production and purification of high-quality functional ATX for research and practical application of multiple functional motogen, ATX/NPP-2.

• Fisheries and Aquatic Sciences
• /
• v.25 no.3
• /
• pp.140-150
• /
• 2022

A method of ultrahigh performance liquid chromatography coupled to high resolution mass spectrometry (UPLC-HRMS) was established for characterization of the lipid profile of Skipjack tuna. Over 300 lipid molecular species were identified through cross-acquisition in both positive and negative ion mode. Phospholipids (PLs) were dominant in Skipjack tuna. Lysophosphatidylethanolamine (LPE), phosphatidylethanolamine (PE), lysophosphatidylcholine (LPC) and phosphatidylcholine (PC) were the main lipid molecular species in PLs, accounting for 89.24% of the total PLs. The ratio of sphingolipids (SLs) and glycerolipids (GLs) were considerable, accounting for 12.30% and 13.60% of the total lipids respectively. Ceramide (Cer) was the main lipid molecular species of SLs, accounting for 64.96% of total SLs, followed by sphingomyelin (SM), accounting for 25.45% of total SLs. Ether diglycerides (ether DG) were the main lipid molecular species of GLs (97.83%). The main fatty acids (FAs) are unsaturated fatty acids (UFAs) in Skipjack tuna. Besides, a new FAs class branched fatty acid esters of hydroxy fatty acids (FAHFA) was detected, together with the FA. The active lipids identified in this study can be used to evaluate the nutritional value of Skipjack tuna.

• IMMUNE NETWORK
• /
• v.23 no.4
• /
• pp.28.1-28.20
• /
• 2023

Lipid accumulation in macrophages is a prominent phenomenon observed in atherosclerosis. Previously, intimal foamy macrophages (FM) showed decreased inflammatory gene expression compared to intimal non-foamy macrophages (NFM). Since reprogramming of lipid metabolism in macrophages affects immunological functions, lipid profiling of intimal macrophages appears to be important for understanding the phenotypic changes of macrophages in atherosclerotic lesions. While lipidomic analysis has been performed in atherosclerotic aortic tissues and cultured macrophages, direct lipid profiling has not been performed in primary aortic macrophages from atherosclerotic aortas. We utilized nanoflow ultrahigh-performance liquid chromatography-tandem mass spectrometry to provide comprehensive lipid profiles of intimal non-foamy and foamy macrophages and adventitial macrophages from Ldlr^-/- mouse aortas. We also analyzed the gene expression of each macrophage type related to lipid metabolism. FM showed increased levels of fatty acids, cholesterol esters, phosphatidylcholine, lysophosphatidylcholine, phosphatidylinositol, and sphingomyelin. However, phosphatidylethanolamine, phosphatidic acid, and ceramide levels were decreased in FM compared to those in NFM. Interestingly, FM showed decreased triacylglycerol (TG) levels. Expressions of lipolysis-related genes including Pnpla2 and Lpl were markedly increased but expressions of Lpin2 and Dgat1 related to TG synthesis were decreased in FM. Analysis of transcriptome and lipidome data revealed differences in the regulation of each lipid metabolic pathway in aortic macrophages. These comprehensive lipidomic data could clarify the phenotypes of macrophages in the atherosclerotic aorta.

• Korean Journal of Food Science and Technology
• /
• v.26 no.2
• /
• pp.123-126
• /
• 1994

Changes in lipid components of pollack meat during sun-drying and effects of NaCl on lipid oxidation were examined. TBA values and peroxide values of sun dried pollack(SD), salted and sun dried pollack (SS) were 0.142 and 14.8 meq/kg, 0.226 and 20.0 meq/kg after sun-drying, respectively. Raw pollack contained 6.12% total lipid consisted of 2.42% neutral lipid(NL) and 3.70% phospholipid(PL) as dry basis, and there were $47{\sim}65%$ decrease in PL content during sun-drying. The NL class of raw pollack mainly consisted of triglyceride(TG), sterol(ST)+diglyceride(DG), hydrocarbon(HC)+sterol ester(SE), and main components in PL class were phosphatidylcholine(PC), phosphatidylethanolamlne(PE) and phosphatidylserine(PS). The contents of TG, ST+DG, PC and PE decreased, while those of free fatty acid, HC+SE, sphingomyelin and lysophosphatidylcholine increased markedly during sun-drying. The major fatty acids of TL in raw pollack, PD and SD samples were generally 22:6, 16:0, 20:5, 18:1 and 18:3; 20:5 decreased markedly during sun-drying, while saturates and monoenes such as 16:0, 18:0 and 18:1 increased slightly. And remaining ratios of polyunsaturated fatty acids of TL, NL and PL in SD and SS samples were 81.1%, 92.5%. 73.3%, and 74.6%, 74.1%, 45.4%, respectively. The results of changes in lipid components during sun-drying showed that sodium chloride catalyzed the lipid oxidation of pollack meat during drying processing.

• The Korean Journal of Physiology and Pharmacology
• /
• v.17 no.3
• /
• pp.181-187
• /
• 2013

Reactive oxygen species (ROS) are generated in various cells, including vascular smooth muscle and endothelial cells, and regulate ion channel functions. $K_{Ca}3.1$ plays an important role in endothelial functions. However, the effects of superoxide and hydrogen peroxide radicals on the expression of this ion channel in the endothelium remain unclear. In this study, we examined the effects of ROS donors on $K_{Ca}3.1$ expression and the $K^+$ current in primary cultured human umbilical vein endothelial cells (HUVECs). The hydrogen peroxide donor, tert-butyl hydroperoxide (TBHP), upregulated $K_{Ca}3.1$ expression, while the superoxide donors, xanthine/xanthine oxidase mixture (X/XO) and lysophosphatidylcholine (LPC), downregulated its expression, in a concentration-dependent manner. These ROS donor effects were prevented by antioxidants or superoxide dismustase. Phosphorylated extracellular signal-regulated kinase (pERK) was upregulated by TBHP and downregulated by X/XO. In addition, repressor element-1-silencing transcription factor (REST) was downregulated by TBHP, and upregulated by X/XO. Furthermore, $K_{Ca}3.1$ current, which was activated by clamping cells with 1 ${\mu}M$ $Ca^{2+}$ and applying the $K_{Ca}3.1$ activator 1-ethyl-2-benzimidazolinone, was further augmented by TBHP, and inhibited by X/XO. These effects were prevented by antioxidants. The results suggest that hydrogen peroxide increases $K_{Ca}3.1$ expression by upregulating pERK and downregulating REST, and augments the $K^+$ current. On the other hand, superoxide reduces $K_{Ca}3.1$ expression by downregulating pERK and upregulating REST, and inhibits the $K^+$ current. ROS thereby play a key role in both physiological and pathological processes in endothelial cells by regulating $K_{Ca}3.1$ and endothelial function.

• Journal of Ginseng Research
• /
• v.43 no.2
• /
• pp.209-217
• /
• 2019

Background: Ginseng is a traditional herbal medicine for human health. Ginseng contains a bioactive ligand named gintonin. The active ingredient of gintonin is lysophosphatidic acid C18:2 (LPA C18:2). We previously developed a method for gintonin-enriched fraction (GEF) preparation to mass-produce gintonin from ginseng. However, previous studies did not show the presence of other bioactive lipids besides LPAs. The aim of this study was to quantify the fatty acids, lysophospholipids (LPLs), and phospholipids (PLs) besides LPAs in GEF. Methods: We prepared GEF from white ginseng. We used gas chromatography-mass spectrometry for fatty acid analysis and liquid chromatography-tandem mass spectrometry for PL analysis, and quantified the fatty acids, LPLs, and PLs in GEF using respective standards. We examined the effect of GEF on insulin secretion in INS-1 cells. Results: GEF contains about 7.5% linoleic (C18:2), 2.8% palmitic (C16:0), and 1.5% oleic acids (C18:1). GEF contains about 0.2% LPA C18:2, 0.06% LPA C16:0, and 0.02% LPA C18:1. GEF contains 0.08% lysophosphatidylcholine, 0.03% lysophosphatidylethanolamine, and 0.13% lysophosphatidylinositols. GEF also contains about 1% phosphatidic acid (PA) 16:0-18:2, 0.5% PA 18:2-18:2, and 0.2% PA 16:0-18:1. GEFmediated insulin secretion was not blocked by LPA receptor antagonist. Conclusion: We determined four characteristics of GEF through lipid analysis and insulin secretion. First, GEF contains a large amount of linoleic acid (C18:2), PA 16:0-18:2, and LPA C18:2 compared with other lipids. Second, the main fatty acid component of LPLs and PLs is linoleic acid (C18:2). Third, GEF stimulates insulin secretion not through LPA receptors. Finally, GEF contains bioactive lipids besides LPAs.

• Journal of Life Science
• /
• v.34 no.3
• /
• pp.179-190
• /
• 2024

Cannabis sativa is a plant widely cultivated worldwide and has been used as a material for food, medicine, building materials and cosmetics. In this study, we assessed the functional effects of C. sativa stem and root extracts using network pharmacology and confirmed their novel functions. The components in stem and root ethanol extracts were identified by gas chromatography-mass spectrometry analysis, and networks between the components and proteins were constructed using the STICHI database. Functional annotation of the proteins was performed using the KEGG pathway. The effects of the extracts were confirmed in lysophosphatidylcholine-induced THP-1 cells using real-time PCR. A total of 21 and 32 components were identified in stem and root extracts, respectively, and 147 and 184 proteins were linked to stem and root components, respectively. KEGG pathway analysis showed that 69 pathways, including the MAPK signaling pathway, were commonly affected by the extracts. Further investigation using pathway networks revealed that terpenoid backbone biosynthesis was likely affected by the extracts, and the expression of the MVK and MVD genes, key proteins in terpenoid backbone biosynthesis, was decreased in LPC-induced THP-1 cells. Therefore, this study determined the diverse function of C. sativa extracts, providing information for predicting and researching the effects of C. sativa.