• 생명과학회지
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• 제24권7호
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• pp.743-749
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• 2014

Orotic acid 1% 수준으로 지방간을 유발시킨 흰쥐에 Monascus purpureus 균주 발효 당귀 5%를 식이 첨가하여 간 조직 및 혈중의 지질대사에 미치는 영향을 검토하였다. 발효 참당귀 분말을 10일간 식이 급여한 결과, orotic acid 유발 지방간 흰쥐의 간 조직 내 중성지질 농도를 낮추고, 혈청 지질 농도가 정상 수준으로 회복되었음을 확인하였다. 간 조직 내 MTP mRNA 및 PDI mRNA 발현 정도는 정상군에 비해 발효 당귀군에서 약간 증가하는 경향을 보이거나, 유의적으로 증가된 수치를 나타내었다. 간 조직의 형태학적 관찰에서는 세포 간극이 좁고 잘 구성된 소엽구조를 하고 있는 정상군에 반해 OA 대조군은 orotic acid 투여로 소포성 지방 변성이 소엽 중심대에 나타나 있고, 지방세포의 크기와 수가 증가하여 전형적인 지방간 형태의 양상을 보였다. 한편, 발효 당귀군의 간장은 간세포의 소포성 지방세포들이 현저히 감소함으로써, 정상 수준의 형태학적 간 성상을 나타내어 지방간 개선에 긍정적인 영향을 미치는 것으로 판단되며, 간 조직의 지질 축적 억제로 인한 지질대사 개선에도 효과가 있는 것으로 사료된다.

• Journal of Nutrition and Health
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• 제30권6호
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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.

• Journal of Nutrition and Health
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• 제30권8호
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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.

• Asian-Australasian Journal of Animal Sciences
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• 제30권5호
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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.

• Molecules and Cells
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• 제19권3호
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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.

• 한국수정란이식학회지
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• 제13권3호
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• pp.277-289
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• 1998

정자의 무분별한 이동을 충분히 억제하면서 운동성을 저해하지 않는 sucrose 층으로부터의 swim-up 분리체계를 구축하기 위하여, 정자의 이동에 장애가 될 수 있는 sucrose 층에 첨가되는 sucrose수준과 정자의 이동과 운동을 극대화시킬 수 있는 배양시간 및 sucrose 이중층의 형태를 조사하였던 바, 얻어진 결과는 다음과 같다. 1. 10mM의 sucrose 층은 정자의 swim-up 이동을 억제하였다. 2. Sucrose 층을 이용하지 않은 swim-up 분리에서 25분간 배양후 회수된 정자의 수가 유의하게 증가되었다. 한편, sucrose 층을 이용한 swim-up 분리에서 10분간 배양후 회수된 정자의 수가 유의하게 증가되었으며 정자의 운동성도 유지되었다 3. Sucrose 이중층 Type I과 Type II 간에 장애효과에는 유의한 차이는 없었으나, 장애공간이 짝은 Type II 이중층에서 시료 채취가 편리하였다. 정자의 주화적 이동과 관련하여 난포액의 기능 을 밝히기 위하여, 난포액의 희석과 열처리 및 난포액으로부터 추출한 단백질과 지질이 Type II Sucrose 층으로부터 10분간 배양한 정자의 swim-up 분리에 미치는 효과를 조사하였던 바 얻어진 결과는 다음과 같다 4. 난포액은 정자의 이동과 운동을 자극하였고 수정능획득정자를 유인하였으며, 특히 10% 난포액에서 수정능획득정자의 유인효과가 가장 크게 나타났다. 5. 저온 열처리 (56$^{\circ}C$, 30분)는 정자의 이동과 운동을 자극하는 난포액의 효과를 감소시키지 않았다. 6. 난포액으로부터 추출한 단백질 성분은 정자의 이동을 자극하였으나, 단백질 성분의 여과는 정자의 이동을 자극하는 단백질 성분의 효과를 감소시켰다. 7. 난포액으로부터 추출한 지질 성분은 정자의 이동과 운동을 유의하게 자극하지 않았다. 8. 난포액으로부터 추출된 단백질과 지질의 병용처리는 정자의 이동과 운동을 자극하는 난포액의 효과를 감소시켰다. 결론적으로 정자의 swim-up 분리에 sucrose 이중층을 이용할 수 있으며, 난포액은 sucrose 층으로부터 정자의 이동과 운동을 자극하고 수정능획득정자를 유인하였다. 특히 열내성을 가진 단백질 성분이 정자의 이동을 자극하였다.

• 한국수정란이식학회:학술대회논문집
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• 한국수정란이식학회 2002년도 국제심포지엄
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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.

• 한국수정란이식학회:학술대회논문집
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• 한국수정란이식학회 2002년도 국제심포지엄
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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.

• 한국미생물·생명공학회지
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• 제21권2호
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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.

• 생명과학회지
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• 제22권11호
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• pp.1487-1492
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• 2012

복숭아는 오래 전부터 알레르기를 일으키는 식품으로 알려져 있으나 특정 식품과의 교차반응에 대한 연구는 부족한 실정이다. 본 연구에서는 한국에서 재배되고 있는 복숭아, 앵두, 고추의 단백질을 추출하여 복숭아 알레르기를 가진 환자가 앵두와 고추에 대해 교차반응성을 일으키는지 확인하고 이 단백질들의 소화효소처리에 따른 안정성을 조사하였다. 복숭아, 앵두 및 고추 단백질 추출물은 Tricine-SDS-PAGE상에서 3 kDa부터 26 kDa 이상까지 넓은 범위의 단백질 분포를 보였으나 각기 다른 패턴을 가지는 것을 확인하였다. 복숭아 단백질의 항원성을 확인하기 위해 복숭아 민감성 환자의 혈청과의 IgE반응성을 확인한 결과, 환자 혈청은 모두 복숭아의 주요 항원성 단백질 lipid transfer protein로 예상되는 9 kDa 부근에서 강한 IgE결합력을 보였으며 또한 9 kDa 단백질이 anti-LTP1 polyclonal antibody에 의해 검출되는 것을 확인하였다. 앵두와 고추 단백질 추출물은 23 kDa 부근에서의 IgE결합력이 9 kDa과의 반응성보다 크게 나타났다. 복숭아, 앵두 및 고추의 소화효소에 대한 안정성을 SDS-PAGE 상에서 확인한 결과, 복숭아, 앵두 및 고추의 항원성 단백질은 인공 위액 및 장액에 의해 완전히 분해되지 않아 알레르기를 유발할 잠재성이 있는 것을 확인할 수 있었다. 본 연구 결과를 바탕으로, 복숭아, 앵두 및 고추 단백질 추출물이 소화효소처리에 안정성을 가지는 특성을 가지며 복숭아 알레르기 증상을 가지는 사람에게 앵두 및 고추는 교차반응성을 일으킬 가능성이 크다는 것을 확인할 수 있었다.