• Molecules and Cells
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• 제42권1호
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• pp.45-55
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• 2019

The liver is involved in a wide range of activities in vertebrates and some other animals, including metabolism, protein synthesis, detoxification, and the immune system. Until now, various methods have been devised to study liver diseases; however, each method has its own limitations. In situ liver perfusion machinery, originally developed in rats, has been successfully adapted to mice, enabling the study of liver diseases. Here we describe the protocol, which is a simple but widely applicable method for investigating the liver diseases. The liver is perfused in situ by cannulation of the portal vein and suprahepatic inferior vena cava (IVC), with antegrade closed circuit circulation completed by clamping the infrahepatic IVC. In situ liver perfusion can be utilized to evaluate immune cell migration and function, hemodynamics and related cellular reactions in each type of hepatic cells, and the metabolism of toxic or other compounds by changing the composition of the circulating media. In situ liver perfusion method maintains liver function and cell viability for up to 2 h. This study also describes an optional protocol using density-gradient centrifugation for the separation of different types of hepatic cells, allowing the determination of changes in each cell type. In summary, this method of in situ liver perfusion will be useful for studying liver diseases as a complement to other established methods.

• 한국미생물·생명공학회지
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• 제39권1호
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• pp.9-19
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• 2011

Bacillus subtilis의 pyrimidine biosynthetic (pyr) operon은 UMP의 de nove 생합성에 관여하는 enzyme들을 encode할 뿐만 아니라, 조절단백질인 PyrR도 encode한다. PyrR은 pyr mRNA-binding 조절 기능과 uracil phosphoribosyltransferase activity를 동시에 가지는 bifunctional 단백질이다. 본 연구에서는 Proteomic analysis를 이용하여 Uracil - 환경에서 DB104${\Delta}$pyrR의 단백질 패턴을 분석하여 단백질 레벨에서 PyrR 단백질의 실질적인 조절 양상을 관찰하였다. 두 균주의 세포질 단백질은 다양한 발현의 차이를 보였으며, Silver 염색된 2D-gel의 pI 4~10 사이에서는 1,300여개의 단백질이 검출되었으며, 단백질 발현 차이를 보이는 172개의 spot 중에서 42개의 단백질이 identification 되었다. 그 결과 pyr operon의 단백질(PyrAa, PyrAb, PyrB, PyrC, PyrD, and PyrF)이 모두 Up regulation이 이루어지고 있음을 확인할 수 있었으며, 이것은 단백질 레벨에서 Pyrimidine 생합성 과정이 PyrR에 의해서 정확히 Regulation 되어짐을 확인할 수 있었다. 또한 Pyrimidine 생합성의 Up regulation과 Down regulation 상태의 단백질의 패턴 양상도 분석할 수 있게 되었다. Pyrimidine의 생합성 과정은 DNA를 구성하는 기본적인 구성 요소를 생산하는 과정으로서 여러가지 Metabolism 가운데 중요한 위치를 차지하고 있다. 만약 Pyrimidine의 생합성 과정이 Over- expression된다면 다른 Metabolism의 균형에도 변화가 올 것이다. Proteomics Analysis에 이용한 DB104${\Delta}$pyrR 균주는 Pyrimidine 생합성의 조절에 관여하는 PyrR knock out 균주로서 Uracil - 환경에서는 전체적인 Pyrimidine 생합성 조절이 Up regulation이 되어지므로 Up regulation 동안 어떤 Metabolism에 영향을 주는지 관찰을 할 수 있게 되었다. 특히 Amino Acid Metabolism에 관계있는 단백질의 Up regulation이 이루어짐을 관찰할 수 있었으며 이것은 현재 각광을 받고 있는 단백질 산업에 응용함으로써 산업적으로 많은 기대를 할 수 있을 것으로 예상되어진다.

• Asian-Australasian Journal of Animal Sciences
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• 제33권1호
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• pp.166-176
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• 2020

Objective: An experiment was conducted to determine the effects of L-arginine (L-Arg) and N-carbamoylglutamic acid (NCG) on the growth, metabolism, immunity and community of cecal bacterial flora of weanling and young rabbits. Methods: Eighteen normal-grade male weanling Japanese White rabbits (JWR) were selected and randomly divided into 6 groups with or without L-Arg and NCG supplementation. The whole feeding process was divided into weanling stage (day 37 to 65) and young stage (day 66 to 85). The effects of L-Arg and NCG on the growth, metabolism, immunity and development of the ileum and jejunum were compared via nutrient metabolism experiments and histological assessment. The different communities of cecal bacterial flora affected by L-Arg and NCG were assessed using high-throughput sequencing technology and bioinformatics analysis. Results: The addition of L-Arg and NCG enhanced the growth of weanling and young rabbit by increasing the nitrogen metabolism, protein efficiency ratio, and biological value, as well as feed intake and daily weight gain. Both L-Arg and NCG increased the concentration of immunoglobulin A (IgA), IgM, and IgG. NCG was superior to L-Arg in promoting intestinal villus development by increasing villus height, villus height/crypt depth index, and reducing the crypt depth. The effects of L-Arg and NCG on the cecal bacterial flora were mainly concentrated in different genera, including Parabacteroides, Roseburia, dgA-11_gut_group, Alistipes, Bacteroides, and Ruminococcaceae_UCG-005. These bacteria function mainly in amino acid transport and metabolism, energy production and conversion, lipid transport and metabolism, recombination and repair, cell cycle control, cell division, and cell motility. Conclusion: L-Arg and NCG can promote the growth and immunity of weanling and young JWR, as well as effecting the jejunum and ileum villi. L-Arg and NCG have different effects in the promotion of nutrient utilization, relieving inflammation and enhancing adaptability through regulating microbial community.

• Applied Biological Chemistry
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• 제44권4호
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• pp.211-214
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• 2001

Drosophila melanogaster Schneider line 2(S2)세포의 외래 단백질 발현 시스템을 이용한 외래 단백질의 한시적 발현을 검토하고, 서로 다른 terminator를 이용하였을 때의 단백질 발현 및 mRNA 발현 정도를 검토하였다. 한시적 발현의 경우 transfection agent를 제거하고 36-48시간 동안 배양한 경우, 가장 높은 green fluorescent protein(GEP)의 발현을 보였다. SV4O p(A), SV4O small T-antigen, 인간 gastrin 3'UTR을 terminator로 지니는 발현 벡터시스템에 각각 endostatin유전자를 cloning시킨 뒤 재조합 endostatin의 mRNA의 발현 정도를 비교하였다. 한시적 발현을 시킨 뒤 36시간 후 endostatin의 발현 정도를 비교해 본 결과 SV40 p(A)를 terminator로 사용했을 때 mRNA및 단백질의 발현이 가장 높았다.

• Journal of Ginseng Research
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• 제36권2호
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• pp.153-160
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• 2012

Panax ginseng has long been used as a traditional herbal medicine. More recently, it has received attention for its anti-diabetic and anti-obesity effects in humans and in animal models of type 2 diabetes. In the present study, we tested the hypoglycemic effects of ginseng berry extract in beta-cell-deficient mice and investigated the mechanisms involved. Red (ripe) and green (unripe) berry extracts were prepared and administered orally (100 or 200 mg/kg body weight) to streptozotocin-induced diabetic mice daily for 10 wk. The body weight was measured daily, and the nonfasting blood glucose levels were measured after 5 and 10 wk after administration. Glucose tolerance tests were performed, and the serum insulin levels were measured. The proliferation of beta-cells was measured in vitro. The administration of red or green ginseng berry extract significantly reduced the blood glucose levels and improved the glucose tolerance in beta-cell deficient mice, with the higher doses resulting in better effects. Glucose-stimulated insulin secretion was significantly increased in berry extract-treated mice compared with streptozotocin-induced diabetic control mice. Treatment with ginseng berry extract increased beta-cell proliferation in vitro. Both red berry and green berry extracts improved glycemic control in streptozotocin-induced diabetic mice and increased insulin secretion, possibly due to increased beta-cell proliferation. These results suggest that ginseng berry extracts might have beneficial effects on beta-cell regeneration.

• Food Science and Biotechnology
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• 제15권5호
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• pp.799-804
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• 2006

Ingestion of green tea has been shown to decrease prostaglandin $E_2$ levels in human colorectum, suggesting that tea constituents modulate arachidonic acid metabolism. In the present study, we investigated the effects of four purified green tea catechins, (-)-epicatechin (EC), (-)-epigallocatechin (EGC), (-)-epigallocatechin-3-gallate (EGCG), and (-)-epicatechin-3-gallate (ECG), on the catalytic activity of cytosolic phospholipase $A_2$ ($cPLA_2$) and release of arachidonic acid and its metabolites from intact cells. At $50\;{\mu}M$, EGCG and ECG inhibited $cPLA_2$ activity by 19 and 37%, respectively, whereas EC and EGC were less effective. The inhibitory effects of these catechins on arachidonic acid metabolism in intact cells were much more pronounced. At $10\;{\mu}M$, EGCG and ECG inhibited the release of arachidonic acid and its metabolites by 50-70% in human colon adenocarcinoma cells (HT-29) and human esophageal squamous carcinoma cells (KYSE-190 and 450). EGCG and ECG also inhibited arachidonic acid release induced by A23187, a calcium ionophore, in both HT-29 and KYSE-450 cell lines by 30-50%. The inhibitory effects of green tea catechins on $cPLA_2$ and arachidonic acid release may provide a possible mechanism for the prevention of human gastrointestinal inflammation and cancers.

• 환경생물
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• 제28권2호
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• pp.79-85
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• 2010

To examine the effect of Codium fragile on blood cholesterol and lipid metabolism, hyperlipidemia was induced in experimental animal rats through the administration of a hypercholesterolemic diet. Codium fragile powder was then administered to the rats for 5 weeks, after which, blood biochemical changes such as blood cholesterol, Aspartate Aminotransferase (AST: serum SGOT) and Alanine Aminotransferase (ALT: serum SGPT) enzyme activity, etc. were determined. And histological changes in liver cells were examined using an electron microscope. Codium fragile treatment resulted in a significant reduction of the levels of total cholesterol, blood triglyceride and low-density cholesterol (LDL. Chol) compared to the control rats. In contrast the expression levels of high-density cholesterol (HDL. chol.) were increased. The AST value of the Codium fragile administration group was significantly reduced and the blood ALT value of the Codium fragile group showed a significant decrease in comparison to the negative control group. In summary, this study demonstrated the beneficial possibilities of Codium fragile in improving the abnormality of lipid metabolism caused by liver cell damage and hyperlipidemia.

• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 1997년도 춘계학술대회
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• pp.97-97
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• 1997

It was reported that ATP depletion occurs and accelerates cell damage during ischemia and reperfusion. To determine the mechanism of cell damage, the change of energy metabolism in liver was studied during ischemia/reperfusion. The groups were divided into four categories : sham-operated group, ischemia/reperfusion group, and two types of ATP-MgCl$_2$ treatment groups(one was treated during ischemia and the another during reperfusion). Rats were administered intravenously saline or ATP-MgCl$_2$. Rats were anesthetized and blood vessels in the left and median lobes of the liver were occluded. After 60min of ischemia, the clamp at those vessels were removed. After ischemia, one and five hours after reflow, energy metabolites(ATP, ADP, AMP, inosine, adenosine, hypoxanthine, xanthine) in liver were measured with HPLC. To observe mitochondrial function, aterial keton body ratio in blood and mitochondrial glutamate dehydrogenase activity in liver were measured. And lipid peroxidation was measured to evalutate the involvement of free radicals. In this study, ATP and ADP were catabolized to their metabolites(AMP, inosine, adenosine, hypoxanthine, xanthine) during ischemia and they resynthesized ATP and ADP during reperfusion. But total purine base were not restored to level of normal rat. The main source of resynthesizing ATP and ADP was AMP. In both ATP-MgCl$_2$ treated groups, mitochondrial function was protected and lipid peroxidation was significantly reduced. Our findings suggest that ischemia/reperfusion impairs hepatic energy metabolism.

• Journal of the Korean Wood Science and Technology
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• 제45권2호
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• pp.168-181
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• 2017

Whole cell of Phanerochaete chrysosporium with reducing agent was applied to verify the degradation mechanism of aromatic compounds derived from lignin precisely. Unlike the free-reducing agent experiment, various degraded products of aromatic compounds were detected under the fungal treatment. Our results suggested that demethoxylation, $C_{\alpha}$ oxidation and ring cleavage of aromatic compounds occurred under the catabolic system of P. chrysosporium. After that, degraded products stimulated the primary metabolism of fungus, so succinic acid was ultimately main degradation product of lignin derived-aromatic compounds. Especially, hydroquinone was detected as final intermediate in the degradation of aromatics and production of succinic acid. In conclusions, P. chrysosporium has an unique catabolic metabolism related to the production of succinic acid from lignin derived-aromatic compounds, which was meaningful in terms of lignin valorization.

• Applied Biological Chemistry
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• 제48권4호
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• pp.425-430
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• 2005

[${\beta}-sitosterol$]은 식물 스테롤로서 인간의 전립선암과 대장암 세포의 성장을 억제하고 생체내 콜레스테롤 농도를 감소시킨다. 본 연구에서는 쑥갓세포 배양에서 ${\beta}-sitosterol$ 생산의 최적화 연구를 수행하였다. 그래서 쑥갓(Chrysanthemum coronarium L.)으로부터 캘러스 유도는 NAA와 BAP의 농도가 각각 1 mg/l의 조합에서 최적이었으며 이들 캘러스로부터 현탁배양 세포주를 확립하였다. 현탁 배양시 초기 세포농도 2 g DCW/l에서 조성이 각각 1배인 탄소원(30 mg/l), 질소원(1900 mg/l $KNO_3$, 1650mg/l $NH_4NO_3$), 무기인산원(170 mg/l)을 포함하는 MS 배지에서 ${\beta}-sitosterol$ 생산이 최적으로 나타났다. Shake-flask를 이용한 현탁배양에서 ${\beta}-sitosterol$의 최대 생산량은 $150{\mu}g/g$ DCW이었다. 그리고 공기부유식 생물반응기의 배양에서는 100 cc/ml의 통기량에서 ${\beta}-sitosterol$의 생산이 $142.8{\mu}g/g$ DCW으로 나타났다.