• 생명과학회지
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• 제29권2호
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• pp.256-264
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• 2019

Lymphotoxin ${\beta}$ receptor ($LT{\beta}R$)는 TNF 계열로 림프조직의 미세구조와 기관형성에 중요한 역할을 한다. MLCK와 ROCK는 세포의 stress fiber 형성조절에 관여하는 주요 신호전달자이다. Fibroblastic reticular cell (FRC)에서 $LT{\beta}R$ 자극을 통한 이런 신호전달자들의 관련성을 알아보기 위해 ML-7 (MLCK 저해제)이 사용되었다. ML7 처리된 FRC에서 SF가 완전히 파괴되었고 anti-$LT{\beta}R$ antibody 처리 세포와 유사하게 ML7 처리 FRC에서 응축된 세포형태를 관찰 할 수 있었다. Y27632로 ROCK를 저해 했을 때 FRC의 액틴 세포골격과 세포형태 변화가 유도 되었다. FRC에서 p-MLC가 액틴과 함께 SF 구성성분을 이루었다. FRC세포 추출물로 Rho-guanosine diphosphate (GDP)/guanosine triphosphate (GTP) 교환활성을 확인했다. Agonistic anti-$LT{\beta}R$ antibody로 $LT{\beta}R$을 자극 했을 때 Rho-GDP/GTP 교환활성이 크게 감소했다. MLCK 저해처럼 $LT{\beta}R$ 자극은 MLC의 인산화를 감소시켰다. Agonistic anti-$LT{\beta}R$ antibody-treated FRC에서 세포골격 구성요소인 세포막과 세포골격 링커 역할을 하는 p-ezrin의 인산화는 감소 되었고 b- actin, 그리고 tubulin 발현도 줄었다. 이런 결과는 FRC의 $LT{\beta}R$ 신호전달을 통한 SF 조절에는 MLCK와 ROCK가 관여하고 있다는 것을 알 수 있었다.

• 대한바이러스학회지
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• 제28권4호
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• pp.369-375
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• 1998

In our preliminary study to find antiviral or antitumor agents from Korean natural products, we found that the Shope fibroma virus (SFV) induced fibromas reaching maximum size at $5{\sim}6$ days with spontaneous disappearance at $15{\sim}20$ days after SFV intracutaneous inoculation into Korean domestic rabbits. However, the sizes of fibromas of rabbits at day 5 after virus inoculation were significantly different individually. Assuming that the variation of tumor size was due to either susceptibility or the preexisting antibodies against SFV in the Korean domestic rabbits, the rabbits were checked for the antibodies against SFV by IFAT using SFV infected RK13 cells. The antibody positive rate of normal Korean domestic rabbits was 32.8% and the sizes of the fibromas of the positive rabbits were significantly smaller than those of negative rabbits (p<0.0001). The fibroma sizes were dependent on the antibody titers of rabbits to SFV. The sizes of fibromas after inoculation of SFV into immunized rabbits were about one tenth of those by the first inoculation into normal rabbits. This is the first report on the antibody prevalence against SFV among normal Korean domestic rabbits and it suggest the existence of a wild fibroma virus or related virus in Korea.

• Bulletin of the Korean Chemical Society
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• 제32권1호
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• pp.286-290
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• 2011

Ginsenoside Rg3 was reported to have important biological activities. We demonstrate conjugation and quantification procedures of ginsenoside Rg3 to gold nanoparticles for future biological and medical applications. Ginsenoside Rg3 was conjugated to spherical gold nanoparticles using a bifunctional heptaethylene glycol linker. The sulfhydryl group of heptaethylene glycol was adsorbed onto gold nanoparticles, and carboxylic acid end of heptaethylene glycol was bonded through a hydroxyl group of Rg3 via ester bond formation. The conjugation of Rg3 was characterized with various spectroscopic techniques, high resolution-transmission electron microscopy, and using Rg3 monoclonal antibody. The Rg3- functionalized gold nanoparticles were $4.7{\pm}1.0$ nm in diameter with a surface charge of -4.12 mV. The total number of Rg3 molecules conjugated to a 3.6 mL solution of gold nanoparticle was determined to be $9.5{\times}10^{14}$ corresponding to ~6 molecules of Rg3/gold nanoparticle. These results suggest that ginsenoside Rg3 is successfully conjugated to gold nanoparticles via heptaethylene glycol linker. The quantification was performed by using Rg3 monoclonal antibody without interference of gold's intrinsic color.

• The Korean Journal of Physiology and Pharmacology
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• 제12권5호
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• pp.225-230
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• 2008

Netrins are secreted molecules and involved in axon guidance, cell migration and tumor development. Recent studies revealed that netrins perform novel functions in such processes as epithelial development and angiogenesis without operating through the classical netrin receptors, DCC (Deleted in Colorectal Cancer) and Unc5h. In the present study, we investigated the roles of netrin-1 and its receptors in cell spreading of human glioblastoma cells, and found that netrin-1 haptotactically enhanced fibronectin-induced cell spreading and focal adhesion formation in U373 glioblastoma cells. Netrin-1 binding to the U373 cell membrane was blocked by an antibody against ${\alpha}v$ integrin subunit, but not by an anti-DCC or anti-Unc5h antibody. In addition, enhancement of the fibronectin response by netrin-1 was abrogated by a function blocking antibody against integrin ${\alpha}v{\beta}3$. Since the ${\alpha}v$ subunit of the integrin family plays an important role in the pathophysiological aspects of cell migration, including tumor angiogenesis and metastasis, our data provide important insight into the molecular mechanism of netrin function.

• 대한미생물학회지
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• 제14권1호
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• pp.71-78
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• 1979

Phytohemagglutinia(PHA) 단독투여(單獨投與) 및 PHA와 Cyclophosphamide(CY)의 동시투여(同時投與)가 마우스의 세포성(細胞性) 및 체액성면역(體液性免疫)에 미치는 영향(影響)을 알아보고저 면양적혈구(緬羊赤血球)(SRBC)로 마우스를 면역(免疫)하기 전에 PHA 및 CY를 투여(投與)하여 말초혈액(末梢血液)의 총(總) 백혈구수(白血球數)와 백혈구백분율검사(白血球百分率檢査), SRBC로 유도(誘導)한 족척종창반응검사(足蹠腫脹反應檢査) 및 비장세포(脾臟細胞)의 rosette 형성검사(形成檢査)를 실시(實施)하였으며 아울러 SRBC에 대(對)한 응집소항체가(凝集素抗體價) 및 용혈소항체가(溶血素抗體價)를 측정(測定)하였다. 그 결과(結果) PHA를 투여(投與)한 마우스군(群)의 면역반응(免疫反應)은 대체(大體)로 감소(減少)되었다. CY만을 투여(投與)한 마우스군(群)의 체액성면역반응(體液性免疫反應)은 감소(減少)되었으나 세포성(細胞性) 면역반응(免疫反應)은 증가(增加)되었다. 이에 반(反)하여 CY 투여전(投與前)에 PHA로 마우스를 전처리(前處理)한 경우 체액성면역반응(體液性免疫反應) 뿐만 아니라 세포성(細胞性) 면역반응(免疫反應)도 감소(減少)되었는데 이 면역반응(免疫反應)의 감소(減少)는 PHA를 CY 투여(投與) 5일전(日前)에 전처리(前處理)한 마우스군(群)에서 가장 현저(顯著)하였다.

• Reproductive and Developmental Biology
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• 제36권3호
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• pp.225-230
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• 2012

Interferon induced transmembrane protein-1 (Ifitm-1) has been reported to have an important role in primordial germ cell formation, and it has expressed in female reproductive organ. In the present study, Ifitm-1 gene expression was identified in testes and all part of epididymis using western immunoblot and immunohistochemistry. Interestingly, Ifitm-1 expression was observed on the head of spermatozoa. To investigate the role of Ifitm-1 gene expression in behavior of spermatozoa after acrosome reaction, fresh sperm was incubated with calcium ionophore to induce acrosome reaction, whereas the expression of Ifitm-1 was not altered after the acrosome reaction. Then to identify the effect of Ifitm-1 in sperm motility and other seminal parameters, different concentration of Ifitm-1 antibody was incubated with spermatozoa, and seminal parameters were assessed using computer-assisted semen analysis (CASA). Interestingly, motility, progressive, and VAP were increased in the sperm with Ifitm-1 antibody treated compared to rabbit serum, however other parameters such as straightness were not changed. In order to identify the functional significance of Ifitm-1 in fertilization, capacitated spermatozoa were pre-incubated with anti-Ifitm-1 antibody and subsequently examined the ability to adhere to mouse oocytes. However, any defection or alteration in sperm-egg fusion was not found, Ifitm-1 antibody treated or non-treated spermatozoa showed a normal penetration. Although the precise role of Ifitm-1 in sperm motility and following fertilization need to be elucidated, this study suggests that the activation of Ifitm-1 on the sperm may enhance the motility of spermatozoa in mice.

• Imaging Science in Dentistry
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• 제30권3호
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• pp.159-168
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• 2000

Purpose: To elucidate the effects of the irradiation and calcium-deficient diet on expression of interleukin (IL)-1 during tooth formation of rat molar. Materials and Methods: The pregnant three-week-old Spague-Dawley rats were used for the study. The control group was non-irradiation/normal diet group, and the experimental groups were irradiation/normal diet group and irradiation/calcium-deficient diet group. The abdomen of the rats on the 9th day of pregnancy were irradiated with single dose of 350 cGy, The rat pups were sacrificed on the 14th day after delivery and the maxillae tooth germs were taken. The specimen were prepared to make sections for light microscopy, and some of tissue sections were stained immunohistochemically with anti-IL-l antibody. Results: In the irradiation/normal diet group, dental follicle showed fewer blood vessels, mononuclear cells, and fusions of mononuclear cells than in non-irradiation/normal diet group. Alveolar bone showed a few osteoblasts and osteoclasts. Periodontal ligament showed collagen fibers and fibroblasts with irregularity. Weak immunoreactivity for IL-l was shown in dental follicle, alveolar bone, and periodontal ligament. In the irradiation/calcium-deficient diet group, dental follicle showed sparse cellularity. Alveolar bone showed diminished number of osteoblasts. Periodontal ligament showed irregular collagen fibers and atrophy of cementoblasts and fibroblasts. No immunoreactivity for IL-1 was shown in dental follicle, alveolar bone, and periodontal ligament. Conclusion: Irradiation and calcium-deficient diet seems to cause disturbance of the expression of interleukin-l during tooth formation of rat molar.

• 한국임상약학회지
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• 제8권1호
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• pp.29-34
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• 1998

Murine CD38 is a 42 kDa type II glycoprotein expressed on cell surface of both B and T lymphocytes. CD38 is a multifunctional enzyme that catalyzes the formation and hydrolysis of cyclic adenosine diphosphoribose (cADPR): ADP-ribosyl cyclase activity of CD38 catalyzes the formation of cADPR from NAD and cADPR hydrolase activity of CD38 catalyzes the hydrolysis of cADPR to ADP-ribose (ADPR). And also, CD38 has the catalytic activity of NAD glycohydrolase (NADase) which catalyzes the hydrolysis of catalyzes the formation and hydrolysis of cyclic adenosine diphosphoribose (cADPR): ADP-ribosyl cyclase activity of CD38 catalyzes the formation of cADPR from NAD to ADPR. In this study, we attempted to purify CD38 from mouse lymphocytes by using the immobilized anti-CD38 monoclonal antibody. The single step immuno-affinity column chromatography resulted in homogeneous purification, showing a single protein of 42 kDa on a SDS polyacrylamide gel. We have investigated the effects of various inhibitors on the enzyme activities of the purified CD38. Cibacron blue (0.5 mM) inhibited all three enzyme activities of CD38, NADase, ADP-ribosyl cyclase and cADPR hydrolase activities. ADPR (2 mM) showed inhibitory effect on both cADPR hydrolase activity and NADase, but not on ADP-ribosyl cyclase activity. However, ATP (2 mM) inhibited only cADPR hydrolase activity. $Zn^{2+}$ (1 mM) showed similar inhibitory effect as that of ADPR, but activated cyclase activity These results suggest that CD38 has three different catalytic activity domains which might be differentially regulated by their specific inhibitors.

• BMB Reports
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• 제36권2호
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• pp.201-206
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• 2003

Two lymphoid-specific proteins, RAG1 and RAG2, are required for the initiation of the V(D)J recombination in vitro. The V(D)J cleavage that is mediated by RAG proteins at the border between the coding and signal sequences results in the production of a hairpin at the coding end and a double-stranded break at the signal end. Two hairpin coding ends are re-opened, modified, and sealed; whereas, the signal ends are directly ligated. Here I report that only RAG1 can carry out a distinct endonucleolytic activity in vitro using an oligonucleotide substrate that is tethered by a short single-stranded DNA. The purified RAG1 protein alone formed a nick at the near position to the recombination signal sequence. This endonucleolytic activity was eliminated by immunoprecipitation using the RAG1-specific antibody, and required the 3'-hydroxy group. All of the RAG1 mutants that were incapable of the nick and hairpin formation in the V(D)J cleavage analysis also showed this new endonucleolytic activity. This suggests that the nicking activity that was observed might be functionally different from the nick formation in the V(D)J cleavage.

• 한국수산과학회지
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• 제39권2호
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• pp.94-99
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• 2006

The tadpole larvae of most ascidians have two sensory pigment cells in their brain vesicle. The anterior otolith pigment cell is sensitive to gravity, whereas the posterior ocellus pigment cell responds to light. Besides these two sensory cells, the larvae also possess another type of sensory receptor cell: hydrostatic pressure receptor (Hpr) cells. The Hpr cells have been presumed to sense hydrostatic water pressure, although no functional analysis has been performed. In larvae of the ascidian Halocynthia reretzi, the development of the Hpr cells and their structure in the brain vesicle are poorly understood. To investigate the morphology and formation of the Hpr cells, we established a monoclonal antibody, Hpr-1, that specifically recognizes Hpr cells. The Hpr-1 antigens became detectable in the brain vesicle at the late tailbud stage. Each Hpr cell projected a small globular body, connected by a short stalk, into the lumen of the brain vesicle. The brain vesicle showed remarkable left-right asymmetry. Pigment cells were located on the right side in the lumen of the brain vesicle, whereas Hpr cells were present in the left side. After metamorphosis, the Hpr cells were observed near the rudimental siphons of the juvenile.