• The Korean Journal of Physiology and Pharmacology
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• 제1권1호
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• pp.35-43
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• 1997

Cis-dichlorodiammine platin${\mu}M$II (Cisplatin), an effective chemotherapeutic agent, induces acute renal failure by unknown mechanisms. To investigate direct toxic effects of cisplatin on the renal proximal tubular transport system, LLC-$PK_1$ cell line was selected as a cell model and the sugar transport activity was evaluated during a course of cisplatin treatment. Cells grown to confluence were treated with cisplatin for 60 min, washed, and then incubated for up to 5 days. At appropriate intervals, cells were tested for sugar transport activity using ${\alpha}-methyl-D-[^{14}C]glucopyranoside$ (AMG) as a model substrate. In cells treated with 100 ${\mu}M$ cisplatin, the AMG uptake was progressively impaired after 3 days. The viability of cells was not substantially changed with cisplatin of less than 100 ${\mu}M$, but it decreased markedly with 150 and 200 ${\mu}M$. In cisplatin-treated cells, the $Na^+$ -dependent AMG uptake was drastically inhibited with no change in the $Na^+$ -independent uptake. Kinetic analysis indicated that Vmax was suppressed, but Km was not altered. The $Na^+$ -dependent phlorizin binding was also decreased in cisplatin-treated cells. However, the AMG efflux from preloaded cells was not apparently retarded by cisplatin treatment. These data indicate that the cisplatin treatment impairs $Na^+$ -hexose cotransporters in LLC-$PK_1$ cells and suggest strongly that defects in transporter function at the luminal plasma membrane of the proximal tubular cells constitute an important pathogenic mechanism of cisplatin nephrotoxicity.

• Biomolecules & Therapeutics
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• 제16권4호
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• pp.410-415
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• 2008

Previously, we identified albumin as an inhibitory factor in serum for cell adhesion of T cells such as human Jurkat T and primary cultured human T cells. In the present study, we found that other hematopoietic cell lines including U-937 human monocytes, THP-1 human monocytes, K-562 promyelocytic leukemia cells, and HL-60 human leukemia cells, also adhere to tissue culture flasks when serum is withdrawn, and albumin exerts an inhibitory effect on cell adhesion by those cells, implying that this inhibition is a common phenomenon in hematopoietic cells. Furthermore, we found that cell adhesion is inhibited by antioxidants such as (-)-epigallocatechin- 3-gallate (EGCG), morin, and a-tocopherol. Our results suggest that albumin may inhibit basal cell adhesion of hematopoietic cells and that the oxidative balance in the plasma may be important for cell adhesion of hematopoietic cells in vivo.

• Archives of Pharmacal Research
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• 제20권2호
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• pp.128-137
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• 1997

The effects of cylindan, a polysaccharide isolated from the basidiocarps of Agrocybe cylindracea, on murine sarcoma 180 tumor and murine immune cells were examined after intraperitoneal administration. Cylindan exhibited a marked life extension effect in mice against ascite forms of sarcoma 180 and Lewis lung carcinoma at a dose of 50 mg/kg/day, although it did not show any direct cytotoxicity against sarcoma 180, X5563, and MM46 murine tumor cells. Cylindan increased numbers of bone marrow stem cells as well as peritoneal exudate cells in flow cytometry using monoclonal antibodies. The tumor bearing mice group apparently showed the increase of macrophages and cytotoxic T lymphocytes in mouse spleen cells during the early stage of tumor growth. But during the later stage, the control group decreased immune cells and cylindan restored the decreased immune cells in the tumor bearing mice to the normal level. In non-specific immune response, cylindan stimulated the bacterial phagocytosis and acid phosphatase production in macrophages. It also activated components of the alternative complement pathway and natural killer activity against YAC-1 lymphoma. In number of plasma cells as token of stimulation of the differentiation of B lymphocytes. In cellular immunity, cylindan restored the depressed response of delayed type hypersensitivity in the tumor bearing mice to 60% of the normal level and increased the interleukin-2 (IL-2) responsiveness in the IL-2 dependent CTLL-2 cells. These results suggest that cylindan did not show direct cytotoxic effects on tumor cells but restored the decreased immune response of the tumor bearing mice.

• 한국영양학회:학술대회논문집
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• 한국영양학회 1995년도 추계학술대회 초록
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• pp.11-34
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• 1995

Growth hormone (GH) plays a key role in regulating postnatal growth and can stimulate growth of animals by acting directly on specific receptors on the plasma membrane of tissues or indirectly through stimulating insulin-like growth factor (IGF)-I synthesis and secretion by the liver and other tissues. IGF-I and IGF-Ⅱ are polypeptides with structural similarity with proinsulin that stimulate cell proliferation by endocrine, paracrine and autocrine mechanisms. The initial event in the metabolic action of IGFs on target cells appears to be their binding to specific receptors on the plasma membrane. Current evidence indicates that the mitogenic actions of both IGFs are mediated primarily by binding to the type I IGF receptors, and that IGF action is also mediated by interactions with IGF-binding proteins (IGFBPs). Six distinct IGFBPs have been identified that are characterized by cell-specific interaction, transcriptional and post-translational regulation by many different effectors, and the ability to either potentiate or inhibit IGF actions. Nutritional deficiencies can have their devastating consequence during growth. Although IGF-I is the major mediator of GH's action on somatic growth, nutritional status of an organism is a critical regulator of IGF-I and IGFBPs. Various nutrient deficiencies result in decreased serum IGF-I levels and altered IGFBP levels, but the blood levels of GH are generally unchanged or elevated in malnutrition. Effects of protein, energy, vitamin C and D, and zinc on serum IGF and IGFBP levels and tissue mRNA levels were reviewed in the text. Multiple factors are involved in the regulation of intestinal epithelial cell growth and differentiation. Among these factors the nutritional status of individuals is the most important. The intestinal epithelium is an important site for mitogenic action of the IGFs in vivo, with exogenous IGF-I stimulating mucosal hyperplasia. Therefore, the IGF system appears to provide and important mechanism linking nutrition and the proliferation of intestinal epithelial cells. In order to study the detailed mechanisms by which intestinal mucosa is regulated, we have utilized IEC-6 cells, an intestinal epithelial cell line and Caco-2 cells, a human colon adenocarcinoma cell line. Like intestinal crypt cells analyzed in vivo or freshly isolated intestinal epithelial cells, IEC-6 cells and Caco-2 cells possess abundant quatities of both type Ⅰ and type Ⅱ IGF receptors. Exogenous IGFs stimulate, whereas addition of IGFBP-2 inhibits IEC-6 cell proliferation. To investigate whether endogenously secreted IGFBP-2 inhibit proliferation, IEC-6 cells were transfected with a full-length rat IGFBP-2 cDNA anti-sense expression construct. IEC-6 cells transfected with anti-sense IGFBP-2 protein in medium. These cells grew at a rate faster than the control cells indicating that endogenous IGFBP-2 inhibits proliferation of IEC-6 cells, probably by sequestering IGFs. IEC-6 cells express many characteristics of enterocyte, but do not undergo differentiation. On the other hand, Caco-2 cells undergo a spontaneous enterocyte differentiation. On the other hand, Caco-2 cells undergo a spontaneous enterocyte differentiation after reaching confluency. We have demonstrated that Caco-2 cells produce IGF-Ⅱ, IGFBP-2, IGFBP-3, and an as yet unidentified 31,000 Mr IGFBP, and that both mRNA and peptide secretion of IGFBP-2 and IGFBP-3 increased, but IGFBP-4 mRNA and protein secretion decreased after the cells reached confluency. These changes occurred in parallel to and were coincident with differentiation of the cells, as measured by expression of sucrase-isomaltase. In addition, Caco-2 cell clones forced to overexpress IGFBP-4 by transfection with a rat IGFBP-4 cDNA construct exhibited a significantly slower growth rate under serum-free conditions and had increased expression of sucrase-isomaltase compared with vector control cells. These results indicate that IGFBP-4 inhibits proliferation and stimulates differentiation of Caco-2 cells, probably by inhibiting the mitogenic actions of IGFs.

• 생명과학회지
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• 제26권8호
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• pp.875-880
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• 2016

진핵세포의 원형질막은 외부환경으로부터 세포를 격리하는 물리적인 장벽뿐만 아니라, 선택적 물질수송, 신호전달등 중요한 기능을 수행한다. 원형질막의 세포질쪽 지질층은 주로 Phosphatidylethanolamine (PE), Phosphatidylserine (PS), Phosphatidylinositides (PIs) 등의 인지질로 구성되어 있는데, 다양한 단백질들이 이들과 직접 결합하거나 이들의 성질을 이용해 원형질막으로 위치된다. 본 연구에서는 원형질막의 안쪽 지질층에 존재하는 특정 인지질과 결합하여, 원형질막에 위치되는 단백질을 과발현시켰을 때 나타나는 세포의 모양변화를 분석하였다. 이를 위해 PS와 PI등과의 선택적 결합으로 원형질막에 위치하는 단백질들과 소수성 또는 정전기적 상호작용으로 원형질막으로 위치되는 단백질들을 HEK293T세포에 과발현시켜 보았다. 그 결과, 대조군으로 사용한 EGFP 단백질과 PI4P에 선택적으로 결합하는 EGFP-P4M-SidM단백질의 발현은 세포의 모양변화를 유도하지 않았다. 반면, PI(4,5)P₂에 결합하는 EGFP-PLCδ1(PH), PI(3,4,5)P₃에 결합하는 EGFP-AKT1(PH), PI4P와 PI(4,5)P₂에 결합하는 OSH2(PH)x2-EGFP의 발현은 세포의 크기가 줄어드는 수축현상이 일어나면서, Lamilapodia나 Filopodia가 형성되는 것을 확인할 수 있었다. 반면에, PS결합을 통해 원형질막에 위치되는 Lact-C2-EGFP과 소수성결합에 의해 원형질막에 위치되는 ApPDE4 long-form인 L(N20)-EGFP이나, 정전기적인 결합을 통해 원형질막으로 위치되는 ApPDE4 short-form인 S(N-UCR1-2)-EGFP단백질의 경우는 세포의 크기가 줄어드는 수축현상은 일어나지만 Lamilapodia나 Filopodia와 같은 모양변화는 나타나지 않는 것을 확인할 수 있었다. 본 연구를 통해, 특정 인지질에 결합하여 원형질막에 위치되는 단백질들이 각각의 인지질 결합 특성에 따라 분류 가능한 세포의 모양변화가 일어남을 확인할 수 있었다.

• 대한소아치과학회지
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• 제26권2호
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• pp.399-415
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• 1999

세포내 유리칼슘(free calcium, $Ca^{2+}$)은 세균에서 고등동물에 이르기까지 거의 모든 세포에서 세포 고유작용을 조절하는 중요한 세포내 신호전달체계(signal transduction system)의 매개체이다. 타액선 세포에서 부교감 신경 자극으로 타액분비가 증가될 때에도 세포내 $Ca^{2+}$ 농도 증가가 가장 중요한 역할을 한다. 그러나 췌장(pancreas)의 경우 세포내 $Ca^{2+}$ 이외에도 인접세포를 전기적 화학적으로 연결해주는 gap junction이 외분비 기능을 직접적으로 조절할 가능설이 제시되었다. 타액선 세포에서도 세포막에 고농도의 gap junction이 존재하고 있으며 gap junction을 통해 인접세포들이 전기적, 화학적으로 연계되어 있어 gap junction이 타액선 세포의 기능을 직접적으로 조절할 가능성을 내포하고 있다. 따라서 gap junction이 타액선의 타액분비 작용에도 중요한 역할을 하며 이러한 작용이 세포내 $Ca^{2+}$ 농도를 조절하여 이루어질 것이라는 가정하에 이를 확인하는 실험을 시행하였다. 흰쥐 악하선에서 유리되는 타액양을 측정하기 위해서 악하선으로 혈액을 공급하는 동맥에 가는 관을 삽입하여 생리 식염수를 관류하면서 타액선관을 통해 타액을 채취하였다. 세포내 $Ca^{2+}$ 농도는 분리한 악하선 acini 내에 $Ca^{2+}$ 농도 변화에 민감하게 반응하는 형광물질인 fura-2를 축적시키고 형광 분석기를 사용하여 형광강도를 측정하여 다음과 같은 결과를 얻었다. 1. CCh 투여로 타액 분비가 증가하였을 때 gap junction을 봉쇄하는 약물인 octanol(1 mM)을 투여하면 타액분비가 봉쇄되었으며 이는 가역적 반응이었다. 2. CCh투여로 세포내 $Ca^{2+}$ 농도가 증가하였을 때 1mM octanol을 투여하면 세포내 $Ca^{2+}$ 농도가 CCh 투여전의 상태로 감소되었다. 3. Octanol은 CCh에 의하여 유발된 초기 $Ca^{2+}$ 증가를 억제하지는 못한 반면에 후기 $Ca^{2+}$ 농도를 감소시켰다. 4. 세포막 $Ca^{2+}$ 통로를 열어주는 약물인 thapsigargin($1{\mu}M$)을 투여하여 세포내 $Ca^{2+}$ 농도를 증가시킨 후 1mM octanol을 투여하면 세포내 $Ca^{2+}$ 농도가 thapsigargin 투여 전의 상태로 감소하였다. 5. 2,5-di-tert-butyl-1,4-benzohydroquinone(TBQ)의 투여로 세포막을 통한 $Ca^{2+}$ 농도의 주기적 변동인 $Ca^{2+}$의 oscillation이 유발되었는데, 이때 1mM octanol을 투여한 경우에 $Ca^{2+}$농도의 oscillation이 정지하여 역시 gap junction을 봉쇄하면 TBQ에 의해서 유발된 세포내 $Ca^{2+}$ 농도의 주기적 변동이 사라지고 $Ca^{2+}$ 농도의 감소가 나타남을 확인하였다. 6. Gap junction을 봉쇄하는 또 다른 약물인 glycyrrhetinic acid($100{\mu}M$)도 CCh 자극으로 인한 타액분비를 억제하였다. 이상의 결과로 미루어 gap junction은 흰쥐 악하선 세포로부터의 타액분비 조절에 중요한 역할을 하는데, 이는 gap junction이 세포막 $Ca^{2+}$ 통로를 조절함으로써 수용체 자극으로 유발된 세포내 $Ca^{2+}$ 농도 변화에 영향을 미친 결과인 것으로 추측된다.

• Molecules and Cells
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• 제37권10호
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• pp.753-758
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• 2014

Sorting nexin 9 (SNX9) is a member of the sorting nexin family of proteins and plays a critical role in clathrinmediated endocytosis. It has a Bin-Amphiphysin-Rvs (BAR) domain which can form a crescent-shaped homodimer structure that induces deformation of the plasma membrane. While other BAR-domain containing proteins such as amphiphysin and endophilin have an amphiphatic helix in front of the BAR domain which plays a critical role in membrane penetration, SNX9 does not. Thus, whether and how SNX9 BAR domain could induce the deformation of the plasma membrane is not clear. The present study identified the internal putative amphiphatic stretch in the $1^{st}$ ${\alpha}$-helix of the SNX9 BAR domain and proved that together with the N-terminal helix ($H_0$) region, this internal putative amphiphatic stretch is critical for inducing membrane tubulation. Therefore, our study shows that SNX9 uses a unique mechanism to induce the tubulation of the plasma membrane which mediates proper membrane deformation during clathrinmediated endocytosis.

• 대한수의학회지
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• 제29권2호
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• pp.33-39
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• 1989

Pure separation of various leukocytes is required for the assessment of their roles in immunological and phisiological function. In this study, pure separation of monocytes from canine peripheral blood was attempted. At first, mononuclear cells (PBMC) were separated by ficoll-hypaque gradient method and then monocytes were recovered from PBMC suspensions in sucrose gradient Sol. (PBMC-Sucrose), autologous plasma (PBMC-Plasma) and autologous serum (PBMC-Serum) incubated at $37^{\circ}C$ for 2 hours. 1. In the separation of PBMC by ficoll-hypaque gradient method in canine blood, higher relative centrifugal force (RCF) was required, as high as more than 1,300xg RCF for 40 minutes, for clear formation of PBMC layer than that in human blood as usually used 400xg RCF for 40 minutes. 2. In monocytes-separation from three PBMC suspensions following PBMC separation, recovery-, purity- and viability-rate of monocytes showed better results in PBMC-Plasma and PBMC-Serum than in PBMC-Sucrose suspension, particulary showing better results from PBMC suspensions performed by centrifugation at 1,500xg RCF for 40 minutes.

• 한국진공학회:학술대회논문집
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• 한국진공학회 2016년도 제50회 동계 정기학술대회 초록집
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• pp.295-295
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• 2016

Recently, TCO films with low carrier concentration, high mobility and high work function are proposed beneficial as front electrode in HIT solar cell due to free-carrier absorption in NIR wavelength region and low Schottky barrier height in the front TCO/a-Si:H(p) interface. We report high transmittance and work function zirconium-doped indium tin oxide (ITO:Zr) films with various plasma (Ar/O2 and Ar) conditions. The role of (Ar/O2) plasma was to enhance the work function of the ITO:Zr films whereas the pure Ar plasma based ITO:Zr showed good electrical properties. The RF magnetron sputtered ITO:Zr films with low resistivity and high transmittance were employed as front electrode in HIT solar cells, yield the best performance of 18.15% with an open-circuit voltage of 710 eV and current density of 34.63 mA/cm2. The high work function ITO:Zr films can be used to modify the front barrier height of HIT solar cell.

• 대한두경부종양학회지
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• 제31권2호
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• pp.66-69
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• 2015

$K{\ddot{u}}ttner$ tumor is a benign inflammatory disease of the salivary gland that clinically mimics a malignant tumor because of its presentation as a unilateral painless hard mass. However the precise cause of this disease is still unknown. Pathologically, it is characterized by infiltration of IgG4-positive plasma cells. Also, it is accompanied with an elevated level of plasma IgG4. We report one case of $K{\ddot{u}}ttner$ tumor of the right submandibular gland in a 37 year old woman.