• Title/Summary/Keyword: Lipid transfer protein

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Effect of Fermented Angelica gigas Nakai on Lipid Metabolism in Orotic Acid Model Rats (발효당귀가 Orotic acid 유발 흰쥐 지질 대사에 미치는 영향)

  • Ahn, Hee-Young;Park, Kyu-Rim;Cho, Young-Su
    • Journal of Life Science
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    • v.24 no.7
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    • pp.743-749
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    • 2014
  • The aim of this study was to identify the effect of fermented Angelica gigas Nakai (A. gigas) on lipid metabolism in orotic acid-induced fatty liver model rats. Sprague-Dawley male rats were randomly divided into four dietary groups (n=6 per group): a normal (N) group fed a standard diet only, OA control, OA acid plus 5% (w/w) A. gigas (OAG), and OA plus 5% (w/w) fermented A. gigas (OFAG). OA treatment induced enlargement of the liver and accumulation of hepatic triglycerides. The consum ption of fermented A. gigas reduced triglyceride concentrations in the liver and increased the serum lipid concentrations to normal levels. Furthermore, OA treatment significantly decreased serum triglyc eride concentrations without diminishing mRNA expression of microsomal triglyceride transfer protei n (MTP) and protein disulfide isomerase (PDI). Hepatic MTP mRNA expression increased 1.08-fold in response to OA treatment, despite triglyceride accumulation in the liver relative to that of the normal group. OFAG administration was slightly lower as compared to the OA treatment. This result suggests that MTP mRNA expression is not always correlated with hepatic triglyceride accumulation in the OA-induced fatty liver model. However, PDI mRNA expression was significantly increased in the OAG and OFAG groups (1.62-fold and 1.63-fold, respectively) compared with the normal group. The hepatocytes in the OA group contained numerous large fat droplets. These were slightly reduced in the OFAG group.

The Role of Membranes and Intracellular Binding Proteins in Cytoplasmic Transport of Hydrophobic Molecules : Fatty Acid Binding Proteins and Long Chain Fatty Acids (세포내 소수성 물질 이동에서 막과 세포내 결합단백질의 역살 : 지방산 결합 단밸직과 장쇄 지방산)

  • 김혜경
    • Journal of Nutrition and Health
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    • v.30 no.6
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    • pp.658-668
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    • 1997
  • Path of a small hydrophobic molecule through the aqueous cytoplasma is not linear. Partition may favor membrane binding by several orders of magnitude : thus significant membrane association will markedly decrease the cytosolic transport rate. The presence of high concentration of soluble binding proteins for these hydrophobic molecules would compete with membrane association and thereby increase transport rate. For long chain fatty acid molecules, a family of cytosolic binding proteins collectively known as the fatty acid binding proteins(FABP), are thought to act as intracellular transport proteins. This paper examines the mechanism of transfer of fluorescent antyroyloxy-labeled fatty acids(AOFA) from purified FABPs to phosholipid membranes. With the exception of the liver FABP, AOFA is transferred from FABP by collisional interaction of the protein with a acceptor membrane. The rate of transfer increased markedly when membranes contain anionic phospholipids. This suggests that positively charged residues on the surface of the FABP may interact with the membranes. Neutralization of the surface lysine residues of adipocyte FABP decreased fatty acid transfer rate, and transfer was found to proceed via aqueous diffusion rather than collisional interaction. Site specific mutagenesis has further shown that the helix-turn-helix domain of the FABP is critical for interaction with anionic acceptor membranes. Thus cytosolic FABP may function in intracellular transport of fatty acid to decrease their membranes association as well as to target fatty acid to specific subcellular sites of utilization.

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Mechanism of Fatty Acid Transfer between Fatty Acid Binding Proteins and Phospolipid Model Membranes (지방산 결합단백질과 인지질막 사이의 지방산이동기전)

  • 김혜경
    • Journal of Nutrition and Health
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    • v.30 no.8
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    • pp.930-935
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    • 1997
  • Fatty acid binging proteins(FABP) are distinct but related gene productes which are found in many mamalian cell types. FABP bind long chain fatty acids in vitro. However, their functions and mechanisms of action, in vivo, remain unknown . Also not known is whether all FABP function similaryly in their respective cell types. or whether different FABP have unique functions. The puropose of the present study was to assess whether different members of the FABP family exhibit different structural and function properties. A comparison was made between heart(H-FABP) and liver (L-FABP). The results show that the binding sites of both FABP are hydrophobic in nature, although the L-FABP site is more nonpolar than the H-FABP site. Additionally, the bound ligand experiences less motional constraint within the H-FABP binding site than within the L-FABP binding site. In accordance with these differences in structural properties, it was found that anthroyloxy-fatty acid transfer from H-FABP to membranes is markedly faster than from L-FABP. moreover, the mechanism of fatty acid transfer to phospholipid membranes appears to occur via transient collisional interactions between H-FABP and membranes. In contrast , transfer of fatty acid from L-FABP occurs via an aqueous diffusion mechanism.

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The effects of high-fat diets composed of different animal and vegetable fat sources on the health status and tissue lipid profiles of male Japanese quail (Coturnix coturnix japonica)

  • Donaldson, Janine;Madziva, Michael Taurai;Erlwanger, Kennedy Honey
    • Asian-Australasian Journal of Animal Sciences
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    • v.30 no.5
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    • pp.700-711
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    • 2017
  • Objective: The current study aimed to investigate the impact of high-fat diets composed of different animal and vegetable fat sources on serum metabolic health markers in Japanese quail, as well as the overall lipid content and fatty acid profiles of the edible bird tissues following significantly increased dietary lipid supplementation. Methods: Fifty seven male quail were divided into six groups and fed either a standard diet or a diet enriched with one of five different fats (22% coconut oil, lard, palm oil, soybean oil, or sunflower oil) for 12 weeks. The birds were subjected to an oral glucose tolerance test following the feeding period, after which they were euthanized and blood, liver, breast, and thigh muscle samples collected. Total fat content and fatty acid profiles of the tissue samples, as well as serum uric acid, triglyceride, cholesterol, total protein, albumin, aspartate transaminase, and total bilirubin concentrations were assessed. Results: High-fat diet feeding had no significant effects on the glucose tolerance of the birds. Dietary fatty acid profiles of the added fats were reflected in the lipid profiles of both the liver and breast and thigh muscle tissues, indicating successful transfer of dietary fatty acids to the edible bird tissues. The significantly increased level of lipid inclusion in the diets of the quail used in the present study was unsuccessful in increasing the overall lipid content of the edible bird tissues. Serum metabolic health markers in birds on the high-fat diets were not significantly different from those observed in birds on the standard diet. Conclusion: Thus, despite the various high-fat diets modifying the fatty acid profile of the birds' tissues, unlike in most mammals, the birds maintained a normal health status following consumption of the various high-fat diets.

The Effect of Ethanol on the Physical Properties of Neuronal Membranes

  • Bae, Moon-Kyoung;Jeong, Dong-Keun;Park, No-Soo;Lee, Cheol-Ho;Cho, Bong-Hye;Jang, Hye-Ock;Yun, Il
    • Molecules and Cells
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    • v.19 no.3
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    • pp.356-364
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    • 2005
  • Intramolecular excimer formation of 1,3-di(1-pyrenyl) propane(Py-3-Py) and fluorescence polarization of 1,6-diphenyl-1,3,5-hexatriene (DPH) were used to evaluate the effect of ethanol on the rate and range of lateral and rotational mobilities of bulk bilayer structures of synaptosomal plasma membrane vesicles (SPMVs) from the bovine cerebral cortex. Ethanol increased the excimer to monomer fluorescence intensity ratio (I'/I) of Py-3-Py in the SPMVs. Selective quenching of both DPH and Py-3-Py by trinitrophenyl groups was used to examine the range of transbilayer asymmetric rotational mobility and the rate and range of transbilayer asymmetric lateral mobility of SPMVs. Ethanol increased the rotational and lateral mobility of the outer monolayer more than of the inner one. Thus ethanol has a selective fluidizing effect within the transbilayer domains of the SPMVs. Radiationless energy transfer from the tryptophans of membrane proteins to Py-3-Py was used to examine both the effect of ethanol on annular lipid fluidity and protein distribution in the SPMVs. Ethanol increased annular lipid fluidity and also caused membrane proteins to cluster. These effects on neuronal membranes may be responsible for some, though not all, of the general anesthetic actions of ethanol.

Effect of Follicular Fluid on Sperm Swim-up Separation with Sucrose Layer (난포액이 Sucrose 층을 이용한 정자의 Swim-up 분리에 미치는 효과)

  • 김경화;여영근;박영식
    • Journal of Embryo Transfer
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    • v.13 no.3
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    • pp.277-289
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    • 1998
  • To establish a system for sperm swim-up separation through sucrose layer, indiscreet sperm migration should sufficiently to block but movement of sperm shouldn't inhibit. Thus, the effects of sucrose levels in sucrose layer, incubation times and types of sucrose layer on sperm separation were examined. And the results obtained were as follows; 1. Layer of 10mM sucrose inhibited sperm swim-up migration through sucrose layer. 2. Incubation for 25 minutes without sucrose layer significantly increased sperm swim-up migration. However, incubation for 10 minutes to induce swim-up through sucrose layer significantly stimulated sperm migration and maintained sperm movement. 3. There was no significant difference between Type I and Type II in barrier effect of sucrose layer. However, sucrose layer of Type II with shorter distance of barrier was efficient for sampling. To elucidate a function of follicular fluid on sperm chemotaxis using in vitro system of sucrose layer of Type II and incubation for 10 minutes, the effects of dilution, heat treatment, and protein and lipid extracts of follicular fluid on sperm swim-up separation were examined. And the results obtained were as follows; 4. Follicular fluid stimulated sperm migration and movement, and significantly-attracted capacitated-sperm at 10% level. 5. Follicular fluid heated at 55$^{\circ}C$ for 30 minutes maintained the effect of follicular fluid stimulating sperm migration and movement. 6. Follicular protein stimulated sperm movement that was reduced by filtration of the protein. 7. Follicular lipid didn't significantly stimulate sperm migration and movement. 8. Both of follicular protein and lipid reduced the effect of follicular fluid stimulating sperm migration and movement. In conclusion, sucrose layer could be used for a barrier against indiscreet sperm migration by swim-up. And follicular fluid stimulated migration and movement of sperm and attracted capacitated-sperm through sucrose layer. Especially, heat-resistant protein of follicular fluid stimulated sperm migration.

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Establishment of an Efficient System for the Production of Transgenic Somatic Cell Nuclear Transfer Embryos

  • Cho, J.K.;Bhuiyan, M.M.U.;Jang, G.;Park, E.S.;Chang, K.H.;Park, H.J.;Lim, J.M.;Kang, S.K.;Lee, B.C.;Hwang, W.S.
    • Proceedings of the Korean Society of Embryo Transfer Conference
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    • 2002.11a
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    • pp.75-75
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    • 2002
  • The present study was conducted for the production of transgenic cloned cows by somatic cell nuclear transfer (SCNT) that secrete human prourokinase into milk. To establish an efficient production system for bovine transgenic SCNT embryos, the offset was examined of various conditions of donor cells including cell type, size, and passage number on the developmental competence of transgenic SCNT embryos. An expression plasmid far human prourokinase (pbeta-ProU) was constructed by inserting a bovine beta-casein promoter, a green fluorescent protein (GFP) marker gene, and a human prourokinase target gene into a pcDNA3 plasmid. Three types of bovine somatic cells including two adult cells (cumulus cells and ear fibroblasts) and fetal fibroblasts were prepared and transfected using a lipid-meidated method. In Experiment 1, developmental competence and rates of GFP expression in bovine transgenic SCNT embryos reconstructed with cumulus cells were significantly higher than those from fetal and ear fibroblasts. In Experiment 2, the effect of cellular senescence in early (2 to 4) and late (8 to 12) passages was investigated. No significant differences in the development of transgenic SCNT embryos were observed. In Experient 3, different sizes of GFP-expressing transfected cumulus cells [large (>30 ${\mu}{\textrm}{m}$) or small cell (<30 ${\mu}{\textrm}{m}$)] were used for SCNT. A significant improvement in embryo development and GFP expression was observed when small cumulus cells were used for SCNT. Taken together, these results demonstrate that (1) adult somatic cells could serve as donor cells in transgenic SCNT embryo production and cumulus cells with small size at early passage were the optimal cell type, and (2) transgenic SCNT embryos derived from adult somatic cells have embryonic development potential.

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Effect of Defined KSOM Medium on the Development of 1-antitrypsin Transgenic Nuclear Transfer Bovine Embryos

  • M.M.U. Bhuiyan;J.K. Cho;G. Jang;Park, E.S.;S.K. Kang;Lee, B.C.;W.S. Hwang
    • Proceedings of the Korean Society of Embryo Transfer Conference
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    • 2002.11a
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    • pp.74-74
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    • 2002
  • Production of u 1-antitrypsin ($\alpha$AT) in transgenic cows has a great value in the field of medicine. The present study was conducted to determine the effect of chemically defined KSOM media on in vitro development of bovine transgenic nuclear transfer (NT) embryos. An expression plasmid for human $\alpha$AT was constructed by inserting a bovine beta-casein promoter, a green fluorescent protein (GFP) marker gene, and a human $\alpha$AT target gene into a pcDNA3 plasmid. Cumulus cells as donor nuclei in NT were collected from a Holstein cow and transfected by lipid-mediated method using FuGene6 (Roche Molecular Biochemicals, USA) as reagent. GFP expressed cumulus cells were introduced into recipient oocytes under DIC microscopy equipped with FITC filter set. After electrical fusion and chemical activation, reconstructed embryos were cultured in 1) SOF + 0.8% BSA, 2) KSOM + 0.8% BSA, 3) KSOM + 10% FBS and 4) KSOM +0.01% PVA for 192 h at 39$^{\circ}C$ with 5% $CO_2$, 5% $O_2$ and 90% $N_2$in humidified condition. The development of the embryos was recorded and the GFP expression in blastocyst was determined under FITC filter. The average fusion rate was 73.8% (251/340; n=8). The development rates to 2-4 cells, morula, blastocysts and expression rates in blastocysts varied from 70.3 to 76.5%, 30.2 to 33.8%, 25.4 to 33.8% and 11.8 to 15.6%, respectively. The difference in development and expression rates of embryos among 4 culture groups was not significant (P>0.05). This study indicates that chemically defined KSOM medium is also able to support development of bovine transgenic NT embryos at similar rate of SOF or KSOM supplemented with BSA or serum.

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The Function of Two n-Alkane Inducible Genes (ALIl, POX18Cm) for n-Alkane Assimilating Candida maltosa (Candida maltosa에서 분리된 n-Alkane 유도성 유전자(ALI1, POX18Cm)의 n-Alkane 대사에 있어서의 기능)

  • ;;Masamichi Takagi
    • Microbiology and Biotechnology Letters
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    • v.21 no.2
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    • pp.181-186
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    • 1993
  • The functions of n-alkane inducible genes, ALI1 and POX18Cm isolated from Canida maltosa were investigated, using it's distruptants. As a result, it is suggested that ALI1 is essential for n-alkane assimilation in C. mltosa and it regulates genes related to assimilation of n-alkane (ALI1, P450alk POX18Cm) at transcriptional level. Nuclear localization experiments indicated that ALI1 was located and functioned in the nucleus. POX18Cm is considered as a peroxisomal nonspecific lipid transfer protein gene related to n-alkane assimilation in C. maltosa also regulated by ALI1. But it had no significant effect on n-alkane assimilation in C. maltosa.

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Cross-Reactivity and Digestive Enzyme Stability of Peach, Korean Cherry, and Hot Pepper (복숭아, 앵두, 고추의 교차반응성 및 소화효소안정성)

  • Kim, Eun-Jung;Ko, Yu-Jin;Lee, Gyeong-Ran;Seol, Hui-Gyeong;Kang, Chang-Min;Ryu, Chung-Ho
    • Journal of Life Science
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    • v.22 no.11
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    • pp.1487-1492
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    • 2012
  • Peach (Prunus persica) has been recognized as a food allergen for over 20 years. However, there is little information about cross-reactivity with other foods. The aim of this study was to research cross-reactivity of Korean cherry and hot pepper on patients allergic to peach and its stability by digestive enzyme treatment. Peach, Korean cherry, and hot pepper proteins were extracted and separated by Tricine-SDS-PAGE analysis. The protein extracts had a wide range of molecular weight, from 3 kDa to more than 26 kDa, and displayed different patterns of protein bands on Tricine-SDS-PAGE. Peach allergic patients' sera were used to detect the allergenic protein in three samples. Three peach allergic patients' sera reacted strongly with 9 kDa protein of peach, which was the expected lipid transfer protein (LTP) as the major allergen of peach and was detected with anti-LTP1 polyclonal antibody. However, the reactivity of the 23 kDa protein in Korean cherry and hot pepper protein was stronger than that of the 9 kDa protein. The stability of protein extracts on digestive enzyme treatment was examined using simulated gastric fluids (SGF) and simulated intestinal fluids (SIF), in which digestive enzyme stability is one of the characteristics of allergen potentially causing food allergy. Findings confirmed that allergenic proteins in peach, Korean cherry, and hot pepper were not completely digested by SGF and SIF treatments from results of SDS-PAGE analysis. These results confirmed that Korean cherry and hot pepper might cause cross-reactivity in peach allergic patients, and its allergenic proteins have stability against digestive enzymes.