• Title/Summary/Keyword: protein tissues

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Effects of enamel matrix derivative and titanium on the proliferation and differentiation of osteoblasts (법랑기질유도체를 도포한 타이태늄 표면에서 조골세포의 증식 및 분화)

  • Park, Sang-Hyun;Lee, In-Kyeong;Yang, Seung-Min;Shin, Seung-Yun;Lee, Yong-Moo;Ku, Young;Rhyu, In-Chul;Chung, Chong-Pyoung;Han, Soo-Boo;Choi, Sang-Mook
    • Journal of Periodontal and Implant Science
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    • v.33 no.3
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    • pp.359-372
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    • 2003
  • Among objectives of periodontal therapy. the principal one is the morphological and functional reconstruction of lost periodontal supporting tissues. This includes de novo formation of connective tissue attachment and the regrowth of alveolar bone. The use of enamel matrix derivative(EMD) may be a suitable means of regeneration new periodontal attachment in the infrabony defects. Implant used to replace lost tooth but, implantitis occurred after installation. The purpose of this study was to investigate the effects of EMD on differentiation and growth of osteoblast in titanium disc. Twentyfive millimeter diameter and 1mm thick Ti disc which was coated 25, 50, 100, 200${\mu}g$/ml of EMD(Emdogain(R)) used as experimental group, 25, 50, 100, 200ng/d of rhBMP-2 as positive control group, and no coat as negative control group. A human osteosarcoma cell line Saos-2 was cultured in Ti disc and cell proliferation and Alkaline phosphatase (ALP) activity were measured at 1 and 6 days. PCR was performed at 2 and 8 hours. Semi-quantitative RT-PCR for mRNA expressions of various osteoblastic differentiation markers -type I collagen, ALP, osteopontin, and bone sialoprotein - were performed at appropriate concentrations based upon the results of MTT and ALP assay. Cultured cell-disc complexes were prepared for scanning electron microscopy (SEM) at 2 hour. Data were analyzed using Mann-Whitney and repeated- measures 1-way analysis of variance(SPSS software version 10,SPSS. Chicago. IL). After culture, there was more osteoblast in EMD100${\mu}g$/ml than in EMD50, 200${\mu}g$/ml on day 6. There was significant difference in experimental and positive control group compared control group, as times go by(1 and 6 days). Alkaline phosphatase activity was different significantly in EMD100, 200${\mu}g$/ml and BMP100, 200${\mu}g$/ml on day 6. The results of reverse transcriptase-polymerase chain reaction (RT-PCR) showed that expression of mRNA for ALPase, collagen type I, osteopontin. hone sialoprotein and BMP-2 was detected at 2 hour and 8 hour in EMI 200${\mu}g$/ml subgroup and BMP100ng/ml subgroup. The results of this study suggest that application of enamel matrix derivative on osteoblast attached to titanium surface facilitate the expression of bone specific protein and the differentiation and growth of osteoblast.

Involvement of Peripheral Benzodiazepine Receptor on the Contractility of Canine Trachealis Muscle (기관근의 수축성에 대한 말초성 Benzodiazepine 수용체의 역할)

  • Rhyu, Han-Young;Choi, Hyung-Cheol;Choi, Eun-Mee;Sohn, Uy-Dong;Lee, Kwang-Youn;Kim, Won-Joon;Ha, Jeoung-Hee
    • The Korean Journal of Physiology and Pharmacology
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    • v.1 no.6
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    • pp.769-774
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    • 1997
  • Non-neuronal high affinity binding sites for benzodiazepines have been found in many peripheral tissues including cardiac muscle and vascular smooth muscle, and have been designated as 'peripheral benzodiazepine receptor'. Benzodiazepines have been shown to induce relaxation of the ileal, vesical, and uterine smooth muscles. However, it is still unclear about possible involvement of peripheral benzodiazepine receptor on the contractility of trachealis muscle. This study was performed to investigate the role of the peripheral benzodiazepine receptor on the contractility of canine trachealis muscle. Canine trachealis muscle strips of 15 mm long were suspended in an isolated organ bath containing 1 ml of physiological salt solution maintained at $37^{\circ}C$, and aerated with $95%\;O_2/5%\;CO_2$. Isometric myography was performed, and the results of the experiments were as follows: Ro5-4684, FGIN-1-27 and clonazepam reduced a basal tone of isolated canine trachealis muscle strip concentration dependently, relaxant actions of RoS-4684 and FGIN-1-27 were antagonized by PK11195, a peripheral benzodiazepine receptor antagonist. Flumazenil, a central type antagonist, did not antagonize the relaxant action of Peripheral type agonists. Saturation binding assay of [3H]Ro5-4864 showed a high affinity$(Kd=5.33{\pm}1.27nM,\;Bmax=\;867.3{\pm}147.2\;fmol/mg\;protein)$ binding site on the canine trachealis muscle. Ro 5-4684 suppressed the bethanechol-, 5-hydroxyoyptamine- and histamine- induced contractions. Platelet activating factor (PAF) exerted strong and prolonged contraction in trachealis muscle strip. Strong tonic contraction by PAE was attenuated by Ro 5-4684, but not by WEB 2086, a PAF antagonist. Based on these results, it is concluded that the peripheral benzodiazepine receptor mediates the inhibitory regulation of contractilty of canine trachealis muscle.

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Morphological Changes in the Skin of Hairless Mouse Fed Various Kimchi Diet (김치종류별 식이가 Hairless Mouse 피부의 형태학적 변화에 미치는 영향)

  • Ryu, Bog-Mi;Ryu, Seung-Hee;Yang, Young-Churl;Lee, Yu-Soon;Jeon, Young-Soo;Moon, Gap-Soon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.33 no.9
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    • pp.1469-1475
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    • 2004
  • Skin is the most frequently exposed tissues to oxidative stress from exogenous and endogenous sources. Dietary antioxidants, which suppress oxidative stress including reactive oxygen metabolites, play an important role in protecting skin from deleterious reactive oxygen species. Kimchi containing lots of antioxidative compounds shows anti-aging effect on skin. Therefore the morphologic changes on the skin of hairless mice fed diets containing Korean cabbage, mustard leaf, and buchu kimchi for 16 weeks were determined. Although epidermal thickness was decreased with age, kimchi prevented this thinning of epidermis compared to control group. In kimchi groups, the staining area of cytokeratin was smaller and stratum corneum was thinner than control group. It suggests that various kinds of kimchi diets prevent the increase of keratinization in epidermis with aging. Type Ⅳ collagen, a major structural protein of basement membrane supporting matrices, existed greater amount in kimchi groups than control group, especially in mustard leaf kimchi group. Rough endoplasmic reticulum (RER) of fibroblast was well developed in dermis of Korean cabbage and mustard leaf kimchi groups, which means collagen synthesis at dermis increased in those kimchi groups. This morphological changes of skin suggest that kimchi consumption can retard skin aging due to the presence of antioxidant and anti-aging compounds, especially some components of mustard leaf kimchi may largely affect on the skin rejuvenescence.

MiR-204 acts as a potential therapeutic target in acute myeloid leukemia by increasing BIRC6-mediated apoptosis

  • Wang, Zhiguo;Luo, Hong;Fang, Zehui;Fan, Yanling;Liu, Xiaojuan;Zhang, Yujing;Rui, Shuping;Chen, Yafeng;Hong, Luojia;Gao, Jincheng;Zhang, Mei
    • BMB Reports
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    • v.51 no.9
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    • pp.444-449
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    • 2018
  • Acute myeloid leukemia (AML) is one of the most common hematological malignancies all around the world. MicroRNAs have been determined to contribute various cancers initiation and progression, including AML. Although microRNA-204 (miR-204) exerts anti-tumor effects in several kinds of cancers, its function in AML remains unknown. In the present study, we assessed miR-204 expression in AML blood samples and cell lines. We also investigated the effects of miR-204 on cellular function of AML cells and the underlying mechanisms of the action of miR-204. Our results showed that miR-204 expression was significantly downregulated in AML tissues and cell lines. In addition, overexpression of miR-204 induced growth inhibition and apoptosis in AML cells, including AML5, HL-60, Kasumi-1 and U937 cells. Cell cycle analysis further confirmed an augmentation in theapoptotic subG1 population by miR-204 overexpression. Mechanistically, baculoviral inhibition of apoptosis protein repeat containing 6 (BIRC6) was identified as a direct target of miR-204. Enforcing miR-204 expression increased the luciferase activity and expression of BIRC6, as well as p53 and Bax expression. Moreover, restoration of BIRC6 reversed the pro-apoptotic effects of miR-204 overexpression in AML cells. Taken together, this study demonstrates that miR-204 causes AML cell apoptosis by targeting BIRC6, suggesting miR-204 may play an anti-carcinogenic role in AML and function as a novel biomarker and therapeutic target for the treatment of this disease.

Identification of a Novel Human Zinc Finger Gene, ZNF438, with Transcription Inhibition Activity

  • Zhong, Zhaomin;Wan, Bo;Qiu, Yun;Ni, Jun;Tang, Wenwen;Chen, Xinya;Yang, Yun;Shen, Suqin;Wang, Ying;Bai, Meirong;Lang, Qingyu;Yu, Long
    • BMB Reports
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    • v.40 no.4
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    • pp.517-524
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    • 2007
  • There were many different families of zinc finger proteins that contained multiple cysteine and/or histidine residues and used zinc to stabilize their folds. The classical C2H2 zinc finger proteins were the founding members of this superfamily and were among the most abundant proteins in eukaryotic genomes. C2H2 proteins typically contained several C2H2 fingers that made tandem contacts along the DNA. Here we reported a novel C2H2 type zinc finger gene, ZNF438, which encoded 828 amino acids that formed five zinc finger domains. Bioinformatics analysis revealed that the ZNF438 was mapped to human chromosome 10p11.2 and shared 62% identity with rat and mouse homologues. RT-PCR analysis indicated that it was ubiquitously expressed in 18 human adult tissues. With immunofluorescence assay, it was shown that the exogenous Flag-tagged ZNF438 was located in nucleus of COS-7 cells. To further explore the function of ZNF438, we examined the transcriptional activity of ZNF438 protein by transfecting recombinant pM-ZNF438 into mammalian cells. The subsequent analysis based on the duel luciferase assay system showed that ZNF438 was a transcriptional repressor.

Effect of PEG chain additive on 6,8-dichloro-2-phenylimidazo[1,2-a] pyridineacetamide (CB185) as a TSPO-binding ligand

  • Lee, Won Chang;Lee, Sang Hee;Denora, Nunzio;Laquintana, Valentino;Lee, Byung Chul;Kim, Sang Eun
    • Journal of Radiopharmaceuticals and Molecular Probes
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    • v.5 no.2
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    • pp.89-100
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    • 2019
  • In our previous studies, we developed a 18F-labeled TSPO-binding ligand, named [18F]CB251, which has been proved to be a promising TSPO-binding PET radiotracer for the detection and monitoring of TSPO expression in pathological diseases. (Ki = 0.27 nM for TSPO, 1.96% ID/g of tumor uptake at 1h post-injection) Based on these results, we utilized 6,8-dichloro-2-phenylimidazo[1,2-a]pyridineacetamide analogs, CB185 (1) as a targeting moiety for the selective delivery of probes and anticancer molecules to TSPO-overexpressed tissues. In this study, we designed CB185 derivatives contains different PEG chains (n = 1, 3 and 5) and fluorescence dye (Cy5) to identify the necessary space between a TSPO-binding ligand and an anticancer agent. Three CB185 derivatives (11a-c) which contains Cy5 and PEG chain, were synthesized and the effect of PEG additive on their TSPO-binding affinities were evaluated using in vitro assays. The binding affinity for compounds 11a-c was lower than that of PK11195 (Ki = 3.2 nM), but still characterized by nanomolar binding affinity for TSPO (Ki = 46.5 nM for 11a, 51.0 nM for 11b, and 388.5 nM for 11c). These results showed that the conjugates are characterized by a moderate binding affinity toward TSPO except for compound 11c, which PEG chain consist of five PEG monomers. Our finding might add useful information to decide the appropriate PET chain length for developing new TSPO-targeting drug carriers.

DEVELOPMENT OF A BIOACTIVE CELLULOSE MEMBRANE FROM SEA SQUIRT SKIN FOR BONE REGENERATION - A PRELIMINARY RESEARCH (멍게와 미더덕 피부의 천연 셀룰로오스 각질을 이용한 골재생 효능을 가진 생활성막의 개발 - 예비 연구)

  • Kim, Soung-Min;Lee, Jong-Ho;Jo, Joung-Ae;Lee, Seung-Cheol;Lee, Suk-Keun
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.31 no.5
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    • pp.440-453
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    • 2005
  • Objectives : To develop a bioactive membrane for guided bone regeneration (GBR), the biocompatibility and bone regenerating capacity of the cellulose membrane obtained from the Ascidians squirt skin were evaluated. Materials and methods : After processing the pure cellulose membrane from the squirt skin, the morphological study, amino acid analysis and the immunoreactivity of the cellulose membrane were tested. Total eighteen male Spraque-Dawley rats (12 weeks, weighing 250 to 300g) were divided into two control (n=8) and another two experimental groups (n=10). In the first experimental group (n=5), the cellulose membrane was applicated to the 8.0 mm sized calvarial bone defect and the same sized defect was left without cellulose membrane in the first control group (n=4). In the another experimental group (n=5), the cellulose membrane was applicated to the same sized calvarial bone defect after femoral bone graft and the same sized defect with bone graft was left without cellulose membrane in the another control group (n=4). Each group was sacrificed after 6 weeks, the histological study with H&E and Masson trichrome stain was done, and immunohistochemical stainings of angiogenin and VEGF were also carried out. Results : The squirt skin cellulose showed the bio-inductive effect on the bone and mesenchymal tissues in the periosteum of rat calvarial bone. This phenomenon was found only in the inner surface of the cellulose membrane after 6 weeks contrast to the outer surface. Bone defect covered with the bioactive cellulose membrane showed significantly greater bone formation compared with control groups. Mesenchymal cells beneath the inner surface of the bioactive cellulose membrane were positive to the angiogenin and VEGF antibodies. Conclusion : We suppose that there still remains extremely little amount of peptide fragment derived from the basement membrane matrix proteins of squirt skin, which is a kind of anchoring protein composed of glycocalyx. This composition could prevent the adverse immunological hypersensitivity and also induce bioactive properties of cellulose membrane. These properties induced the effective angiogenesis with rapid osteogenesis beneath the inner surface of cellulose membrane, and so the possibilities of clinical application in dental field as a GBR material will be able to be suggested.

Pharmacokinetics of Recombinant Human Granulocyte Colony Stimulating Factor (rhG-CSF) Following Intravenous, Intramuscular and Subcutaneous Administration of HM10411 and Filgrastim to Rats and Mice (인과립구 콜로니 자극인자 제제인 HM10411와 필그라스팀의 정맥, 근육 및 피하 주사시 흰쥐와 마우스에서의 약물 동태)

  • Kim, In-Wha;Lee, Sang-Hoon;Kim, Young-Min;Jung, Sung-Youb;Kwon, Se-Chang;Lee, Gwan-Sun;Chung, Suk-Jae;Shim, Chang-Koo
    • Journal of Pharmaceutical Investigation
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    • v.31 no.2
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    • pp.89-94
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    • 2001
  • The pharmacokinetics of recombinant human granulocyte colony stimulating factor (rhG-CSF) following intravenous (i.v.), intramuscular (i.m.) and subcutaneous (s.c.) administration of HM1041l-lyo and HM10411-liq (lyophilized and liquid formulations of rhG-CSF, recently under development by Hanmi Pharmaceutical Company) were studied in rats, and compared with that of Filgrastim (conventional formulation of rhG-CSF on market). The plasma concentration of rhG-CSF was quantified using a specific ELISA. The pharmacokinetic parameters of rhG-CSF, after i.v., i.m. and s.c. administration of Filgrastim, HM1041l-lyo and HM1041l-liq to rats at a rhG-CSF dose of $10\;{\mu}g/kg$, were almost identical among the three formulations. No dose-dependency was observed in the pharmacokinetic parameters of rhG-CSF following i.v. administration in the dose range of $5{\sim}100\;{\mu}g/kg$. rhG-CSF, after i.v. administration of the three preparations at a dose of $10\;{\mu}g/kg$ to rats, was detected at low levels in all of the body tissues with highest tissue/plasma ratio of $0.46{\sim}0.51$ for the kidney at 30 min after the administration. The pharmacokinetics of rhG-CSF, after i.v. administration to mice at a dose of $10\;{\mu}g/kg$, were comparable among the three formulations. In conclusion, HM10411-lyo and HM10411-liq exhibited similar pharmacokinetics for rhG-CSF with Filgrastim regandless of animal species. Considering the fact that HM10411 series, contrary to Filgrastim, are proteins lacking a methionine residue, the methionine moiety in rhG-CSF molecule does not appear to influence the pharmacokinetics of the protein significantly.

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A Protective Effect of Chlorella Supplementation on Cadmium-induced Nephrotoxicity in the Rats

  • Hwang Yoo-Kyeong;Choi Hyun-Jin;Nan Meng;Yoo Jai-Du;Kim Yong-Ho
    • Biomedical Science Letters
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    • v.12 no.1
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    • pp.29-33
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    • 2006
  • The uptake of cadmium in animals is mainly accumulated in and affected to the liver and kidney by binding with red blood cells and serum albumin. The process accounts for more than 50% of the total accumulated cadmium in the body. The kidneys may be damaged without regarding the pathway uptake of cadmium. In a group of rats on supplements of 1% chlorella and 40 ppm cadmium, the concentration of cadmium in urine greatly decreased by 66% compared to control group, and the total synthesis of metallothionein decreased by 48.6% compared to control group. However, no previous study has assessed the protective effect on kidney damage induced by cadmium uptake through supplementation with chlorella. This study analyzed the biochemical marker for kidney damage in the rats after uptake of 40 ppm $CdCl_2$ and supplementation of the diet of Sprague Dawley (SD) rats with 1%, 5%, and 10% chlorella during 4 weeks. In a group of SD rats on supplementation with 1% chlorella and uptake of 40 ppm $CdCl_2,\;\beta_2$ microglobulin in the urine was found to be $3.1\pm0.6\;{\mu}g/L$, a decrease of 58% compared to a group of Sp rats on uptake of $CdCl_2$ only, in which the $\beta_2$ microglobulin was found to be $4.9\pm0.7\;{\mu}g/L$. According to the results of histopathological observation, the accumulation of mild and localized chronic inflammatory cells in kidney tissues was observed in 50% of the SD rats on uptake of cadmium only. In contrast, only 30% of the SD rats on supplementation with 1% chlorella and uptake of 40ppm $CdCl_2$, representing a histopathological abnormality, and there were no histopathological abnormalities at all in groups of SD rats on supplementation with 5% or 10% chlorella and uptake of 40 ppm $CdCl_2$. In conclusion, protein, calcium, and iron, which account for more than 50% of the total dried chlorella composition, may contribute to the reduction nephrotoxicity by stimulating both inhibited absorption of cadium and increased excretion of accumulated cadmium in kidneys.

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Agronomic Characteristics and Aromatic Compositions of Korean Wild Codonopsis lanceolata Collections Cultivated in Field (한국 야생더덕 수집종의 노지 재배시 생육 특성과 향기성분 조성)

  • 이승필;김상국;민기군;조지형;최부술;이상철;김길웅
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.41 no.2
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    • pp.188-199
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    • 1996
  • The native ecological environment and aromatic constituents of Korean wild Codonopsis lanceolata and one Japanese strain were investigated to find Codonopsis lanceolata strains showing high aromatics, and to know regional differences among these strains. The results were as follows : There were no remarkable differences among the Korean wild C. lanceolata strains in ecological environments. Recovery yield of essential oils was highest in Togyusan strain with 0.009%. Difference in protein band patterns among these strains was not recognized, and peroxidase and esterase pattern changes were appeared in different collected regions at the leaf and root tissues. Major aromatic constituents were 11 kinds of aliphatic alcohols such as trans-2-hexenal, 1-hexanol, cis-3-hexanol, and trans-2-hexanol. And Togyusan strain, Sobaeksan strain, and Kayasan strain have the highest aliphatic alcohols of plant essential oils. In particular, BHT(butylated hydroxytoluene), one of the antioxidants, was detected in Chirisan strains.

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