• Title/Summary/Keyword: growth factor protein

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A Study for the Mechanism of Abnormal Proliferation in Vascular Endothelial Cells using Inhibitors to the Signal Transduction Pathway (신호전달 경로의 저해제를 이용한 혈관 내피세포의 비정상적인 증식 기전에 대한 연구)

  • Bae, Yong Chan;Park, Suk Young;Nam, Su Bong;Herh, Jae Young;Kang, Young Seok
    • Archives of Plastic Surgery
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    • v.33 no.1
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    • pp.5-12
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    • 2006
  • Protein tyrosine kinase(PTK), protein kinase C(PKC), oxidase, as a mediator, take a significant role in signal transduction pathway of angiogenesis. The authors utilized the inhibitors, targeting the formation of three co-enzyme in signal transduction pathway in order to quantify the suppression of abnormal vascular endothelial cell proliferation induced by DMH, to compare the level suppression in each up-regulated growth factors, CTGF, CYR61, $ITG{\beta}1$, FHL2, and to identify the relationship between abnormal cell proliferation and signal transduction pathway. Five groups were established; Control group, Group of DMH, Group of DMH-mixed Herbimycin, inhibitor of protein tyrosine kinase, Group of DMH-mixed Calphostin C, inhibitor of protein kinase C, Group Of Dmh-Mixed 10U Catalase, Inhibitor Of oxidase. The rise of vascular endothelial cell was compared by MTT assay, and four growth factors were analysed with RT-PCR method, at pre-administration, 4, 8, 12, and 24 hours after administration. In comparison of abnormal proliferation of vascular endothelial cell induced by DMH, suppression was noticed in Herbimycin and Calphostin C group, and Calphostin C group revealed higher suppression effect. Nevertheless, Catalase group did not have any suppression. In manifestation of four growth factors, Herbimycin and Calphostin C group presented similar manifestation with control group, except in $ITG{\beta}$. Catalse group had similar manifestation with DMH group in all four growth factors. Abnormal proliferation of vascular endothelial cell induced by DMH have a direct relationship with PTK and PKC, more specifically to PKC. Oxidase was confirmed not to have any relevance.

Feeding ratio affects growth, body composition, and blood chemistry of mandarin fish (Siniperca scherzeri) in recirculating aquaculture system

  • Kim, Yi-Oh;Oh, Sung-Yong;Lee, Who-Seung
    • Fisheries and Aquatic Sciences
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    • v.24 no.6
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    • pp.219-227
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    • 2021
  • The effects of various feeding ratios on the growth, body composition, and blood chemistry of the juvenile mandarin fish Siniperca scherzeri (initial body weight 9.6 g) were examined in recirculating freshwater system equipped with 21, 300 L tanks at 20 fish per tank. The triplicate groups of seven feeding ratios treatments were prepared: 100% (control), 95%, 90%, 85%, 80%, 75%, and 70% of satiation. The feed amount of control group was determined by supplying with apparent satiation and then the feed amounts of the other six feeding groups were determined based on the feed amount of the control group. Fish were hand-fed with test diet (55.4% crude protein) for 10 weeks. Weight gain (WG) and specific growth rate of fish fed to 100% satiation were not significantly (p > 0.05) different from those of fish fed to ≥ 80% satiation but were significantly higher than those of fish fed to 75% and 70% satiation. Feed efficiency, protein efficiency ratio, and protein retention of 100% satiation were not significantly different from those of 95% and 90% satiation but were significantly (p <0.05) lower than ≤ 85% satiation. Condition factor, hepatosomatic index, and coefficient variation were not significantly (p > 0.05) affected by feeding ratio. Whole body composition and contents of hematocrit, glutamic oxaloacetic transaminase, glutamic pyruvic transaminase, glucose, total protein, and high-density lipoprotein cholesterol in blood serum were not significantly (p > 0.05) affected by the feeding ratio; however, content of total cholesterol tended to decrease as the feeding ratio decreased. Using broken-line analysis of WG, it was suggested that the optimum feeding ratio of juvenile mandarin fish, ranging from 9.0 g to 37.0 g, appeared to be 87.7% of satiation without growth inhibition.

Secretory Production of Recombinant Urokinase Kringle Domain in Pichia pastoris

  • Kim, Hyun-Kyung;Hong, Yong-Kil;Park, Hyo-Eun;Hong, Sung-Hee;Joe, Young-Ae
    • Journal of Microbiology and Biotechnology
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    • v.13 no.4
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    • pp.591-597
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    • 2003
  • Human urokinase kringle domain, sharing homology with angiostatin kringles, has been shown to be an inhibitor of angiogenesis, which can be used for the treatment of cancer, rheumatoid arthritis, psoriasis, and retinopathy. Here, the expression of the kringle domain of urokinase (UK1) as a secreted protein in high levels is reported. UK1 was expressed in the methylotrophic yeast Pichia pastoris GS115 by fusion of the cDNA spanning from Ser47 to Lys135 to the secretion signal sequence of ${\alpha}-factor$ prepro-peptide. In a flask culture, the secreted UK1 reached about 1 g/l level after 120h of methanol induction and was purified to homogeneity by ion-exchange chromatography. Amino-terminal sequencing of the purified UK1 revealed that it was cleaved at the Ste13 signal cleavage site. The molecular mass of UK1 was determined to be 10,297.01 Da. It was also confirmed that the purified UK1 inhibited endothelial cell proliferation stimulated by basic fibroblast growth factor, vascular endothelial growth factor, or epidermal growth factor, in a dose-dependent manner. These results suggest that a P. pastoris sytem can be employed to obtain large amounts of soluble and active UK1.

Development of Mouse Preimplantation Embryos in Solubilized Matrigel Media (용해된 Matrigel 첨가 배지에서 착상전 생쥐 배아의 발생)

  • Chung, Byung-Mok;Choo, Hyung-Sik;Kang, Byung-Moon;Gye, Myung-Chan
    • Clinical and Experimental Reproductive Medicine
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    • v.27 no.4
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    • pp.381-385
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    • 2000
  • Objective: To verify the effect of two forms (growth factor and growthfactor-reduced) of solubilized Matrigel on the development in mouse preimplantation embryos. Methods: Late 2-cell stage eggs were cultured through the blastocyst stage in the presence of GF- or GFR-Matrigel (0.5%, v/v). Morphological development, cell number and % apoptotic nuclei of blastocyst were measured by Roecst staining and TUNEL of nuclei. Results: Morphological development, number of cells per embryo was significantly increased in the presence of GF- or GFR-Matrigel. Culture of the embryos in the GF-Matrigel gave the best result. Conclusion: Low concentration of solubilized Matrigel improved development of mouse embryos regardless of growth factor content of the Matrigel. Growth factors and extracellular matrix protein included in the Matrigel synergistically potentiated the development of mouse embryos.

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A Case Report of Guided Bone Regeneration Using a Putty-type Demineralized Bone Matrix (골유도재생술에 대한 putty형 탈회 기질골 이용연구)

  • Jeong, Mi-Ae
    • Proceedings of the KAIS Fall Conference
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    • 2011.12a
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    • pp.330-333
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    • 2011
  • Allomatrix (Wright Medical Tech, Inc., USA), is a newly designed, injectable putty with a reliable demineralized bone matrix(DBM), derived from human bone. The compound contains 86% DBM and other bone growth factors such as bone morphogenic protein (BMP)-2, BMP-4, insulin-like growth factor (IGF)-1, and transforming growth factor (TGF)-${\beta}1$. It has excellent os-teoinduction abilities. In addition, DBM is known to have osteoconduction capacity as a scaffold due to its collagen matrix. This product contains a powder, which is a mix of DBM and surgical grade calcium sulfate as a carrier. A practitioner can blend the powder with calcium sulfate solution, making a putty-type material which has the advantages of ease of handling, better fixation, and no need for a membrane, because it can function as membrane itself. This study reports the clinical and radiographic results of various guided bone regeneration cases using Allomatrix, demonstrating its strong potential as a graft material.

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Novel p104 protein regulates cell proliferation through PI3K inhibition and p27Kip1 expression

  • Han, Seung-Jin;Lee, Jung-Hyun;Choi, Ki-Young;Hong, Seung-Hwan
    • BMB Reports
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    • v.43 no.3
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    • pp.199-204
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    • 2010
  • The protein p104 was first isolated as a binding partner of the Src homology domain of phospholipase C$\gamma$1, and has been shown to associate with p85$\alpha$, Grb2. The ectopic expression of p104 reduced cellular growth rate, which was also achieved with the overexpression of only the proline-rich region of p104. The proline-rich region of p104 has been found to inhibit the colony formation of platelet-derived growth factor BB-stimulated NIH3T3 cells and MCF7 cancer cells on soft agar. Mutagenesis analysis showed that the second and third proline-rich regions are essential for growth control, as well as for interaction with p85$\alpha$. Overexpression of p104 increased the level of the cyclin-dependent kinase inhibitor, $p27^{Kip1}$, and inhibited the activity of phosphoinositide 3-kinase (PI3K). In summary, p104 interacts with p85$\alpha$ and is involved in the regulation of $p27^{Kip1}$ expression for the reduction of cellular proliferation.

Differential gene expression pattern in brains of acrylamide-administered mice

  • Han, Chang-Hoon
    • Korean Journal of Veterinary Research
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    • v.52 no.2
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    • pp.99-104
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    • 2012
  • The present study was performed to evaluate the relationship between the neurotoxicity of acrylamide and the differential gene expression pattern in mice. Both locomotor test and rota-rod test showed that the group treated with higher than 30 mg/kg/day of acrylamide caused impaired motor activity in mice. Based on cDNA microarray analysis of mouse brain, myelin basic protein gene, kinesin family member 5B gene, and fibroblast growth factor (FGF) 1 and its receptor genes were down-regulated by acrylamide. The genes are known to be essential for neurofilament synthesis, axonal transport, and neuroprotection, respectively. Interestingly, both FGF 1 and its receptor genes were down-regulated. Genes involved in nucleic acid binding such as AU RNA binding protein/enoyl-coA hydratase, translation initiation factor (TIF) 2 alpha kinase 4, activating transcription factor 2, and U2AF 1 related sequence 1 genes were down-regulated. More interesting finding was that genes of both catalytic and regulatory subunit of protein phosphatases which are important for signal transduction pathways were down-regulated. Here, we propose that acrylamide induces neurotoxicity by regulation of genes associated with neurofilament synthesis, axonal transport, neuro-protection, and signal transduction pathways.

A small molecule approach to degrade RAS with EGFR repression is a potential therapy for KRAS mutation-driven colorectal cancer resistance to cetuximab

  • Lee, Sang-Kyu;Cho, Yong-Hee;Cha, Pu-Hyeon;Yoon, Jeong-Soo;Ro, Eun Ji;Jeong, Woo-Jeong;Park, Jieun;Kim, Hyuntae;Kim, Tae Il;Min, Do Sik;Han, Gyoonhee;Choi, Kang-Yell
    • Experimental and Molecular Medicine
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    • v.50 no.11
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    • pp.12.1-12.12
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    • 2018
  • Drugs targeting the epidermal growth factor receptor (EGFR), such as cetuximab and panitumumab, have been prescribed for metastatic colorectal cancer (CRC), but patients harboring KRAS mutations are insensitive to them and do not have an alternative drug to overcome the problem. The levels of ${\beta}$-catenin, EGFR, and RAS, especially mutant KRAS, are increased in CRC patient tissues due to mutations of adenomatous polyposis coli (APC), which occur in 90% of human CRCs. The increases in these proteins by APC loss synergistically promote tumorigenesis. Therefore, we tested KYA1797K, a recently identified small molecule that degrades both ${\beta}$-catenin and Ras via $GSK3{\beta}$ activation, and its capability to suppress the cetuximab resistance of KRAS-mutated CRC cells. KYA1797K suppressed the growth of tumor xenografts induced by CRC cells as well as tumor organoids derived from CRC patients having both APC and KRAS mutations. Lowering the levels of both ${\beta}$-catenin and RAS as well as EGFR via targeting the $Wnt/{\beta}$-catenin pathway is a therapeutic strategy for controlling CRC and other types of cancer with aberrantly activated the $Wnt/{\beta}$-catenin and EGFR-RAS pathways, including those with resistance to EGFR-targeting drugs attributed to KRAS mutations.

Development of Purification Process of Recombinant Human Vascular Endotherial Growth Factor (VEGF) using Fusion Protein (융합 단백질을 이용한 재조합 인간 혈관내피세포 성장인자의 정제공정 개발)

  • Sung, Keehyun;Kim, In Ho
    • Korean Chemical Engineering Research
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    • v.55 no.3
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    • pp.369-378
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    • 2017
  • Vascular endotherial growth factor (VEGF) is a potent mitogen that stimulates vascular permeability and angiogenesis and has a potential in therapeutic applications. An industrial production method that provides high yield as well as purity is needed. Researches for various factors of mild solubilization with combination of ubiquitin fusion protein to increase solubility were carried out as well as by changing pH and denaturant concentration. Usage of pET28-a bacteral expression vector in BL21 (DE3) host cell was capable of producing approximately 14 g/L VEGF fusion protein in 20L fermentor. A purification process consisting of four chromatography steps including refolding and digestion with UBP1 resulted in mild solublization under the conditions of 2M urea and pH 10.0 due to ubiquitin fusion tag protein that increases in solubility of target protein VEGF. High yield of refolding and dimerization could be obtained between two step Ni-affinity chromatography. Multimeric and misfolded proteins and endotoxin were removed by DEAE anion exchange chromatography. Final monomers were removed from dimers by gel filtration chromatography. Characterization analysis of purified dimeric VEGF was performed using SDS-PAGE and RP-HPLC with a purity of 97%.

The combination effects of PDGF and IGF-I on the proliferation and cellular activity of periodontal ligament cells (PDGF와 IGF-I 병용 사용시 치주인대세포의 증식과 세포활성에 미치는 영향에 관한 연구)

  • Suh, Jo-Young;Shin, Hong-In;Kyung, Hee-Moon
    • Journal of Periodontal and Implant Science
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    • v.26 no.2
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    • pp.396-413
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    • 1996
  • Current acceptable methods for promoting periodontal regeneration are based on removal of diseased soft tissue. root treatment, guided tissue regeneration, introduction of new graft materials and biological mediators. Insulin-like growth factor-I(IGF-I) and Platelet-derived growth factor-BB(PDGF-BB), the members of the polypeptuyde growth factor family have been reported as the biological mediators which regulate a variety cellular matrix biologic activities of wound healing process including the cell proliferation, migration and extracellular matrix synthesis.The purposes of this study is to evaluate the combination effects of IGF-I and PDGF-BB on the cellular activity of the periodontal ligament cells to act as a regeneration promoting agent of periodontal tissue. Human periodontal ligament cells were prepared from the first premolar tooth extracted for the orthodontic treatment and were cultured in DMEM containing 10% FBS at the $37^{\circ}C$, 5% CO2 incubator. Author measured the DNA synthetic activity, and total protein, collagen and noncollagenous protein synthetic activities according to the concentration of 10,100ng/ml IGF-I and1,10 ng/ml PDGF-BB in combination. The results were as follows: Significantly increased in the 1 ng/ml PDGF-BB alone compared to the 10 ng/ml PDGF-BB alone(P<0.01) and in the 1 ng/ml PDGF-BB and 10, 100ng/ml IGF-I in combination compared to the 1 ng/ml PDGF-BB alone(P<0.05, P<0.0l). The synthetic activity of the total protein and collagen is significantly increased like to the synthetic activity of the DNA(P<0.05). The synthetic activity of the noncollagenous protein is increased according to the concentration of IGF_I, but not statistically statistically significant(P>0.05). The percent of the collagen is significantly in the 1ng/ml PDGF-BB and 10ng/ml IGF-I in combination compared to the 1ng/ml PDGF-BB alone(P<0.05) and in the 10ng/ml IGF-I in combination compared to the 10ng/ml PDGF-BB alone(P<0.05). The synthetic activity of the DNA is In conclusions, the percent study shows that PDGF-BB and IGF-I in combination have a potentiality to enhance the DNA synthesis and the total protein and collagen synthesis of The periodontal ligament cells, especially it is more significant in the low concentration of PDGF-BB compared to the high one. Thus, the PDGF-BB and IGF-I in combination may have important roles in promotion of periodontal litgment healing, and consequently, may useful for clinical application in periodontal regenerative procedures.

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