• Title/Summary/Keyword: 단백질 추출

Search Result 1,882, Processing Time 0.051 seconds

Preparation and Characterization of Demineralized Bone Particle-loaded PLGA Scaffold for Tissue Engineered Bone (조직공학적 골재생을 위한 탈미넬화된 골분을 함유한 다공성 지지체의 제조 및 그 특성)

  • Jang Ji Wook;Lee Bong;Han Chang Whan;Kim Mun Suk;Cho Sun Hang;Lee Hai Bang;Khang Gilson
    • Polymer(Korea)
    • /
    • v.28 no.5
    • /
    • pp.382-390
    • /
    • 2004
  • One of the significant natural bioactive materials is demineralized bone particle (DBP) whose has a powerful induce. of new bone growth. In this study, we developed the DBP loaded poly-lactide (PLA) and poly(L-lactide-co-glycolide) (PLGA) scaffolds for the possibility of the application of the tissue engineered bone. PLA/DBP and PLGA/DBP scaffolds were prepared by solvent casting/salt leaching method and were characterized by porosimeter, scanning electron microscopy. BMSCs were stimulated by osteogenic medium and characterized by histological stained Wright-Giemsa, Alizarin red, von Kossa, and alkaline phosphate activity (ALP). DBP impregnated scaffolds with BMSCs were implanted into the back of athymic nude mouse to observe the effect of DBP on the osteoinduction compared with control scaffolds. It can be observed that the porosity was above $90.2\%$ and the pore size was above 69.1$\mu$m. BMSCs could be differentiated into osteoprogenitor cells as result of wright-giemsa, alizarin red, von Kossa and ALP staining. In in vivo study, we could observed calcification region in PLA/DBP and PLGA/DBP groups, but calcification did not occur almost in control scaffolds. From these results, it seems that DBP as well as BMSCs play an important role for bone induction in PLA/DBP and PLGA/DBP scaffolds.

Effects of Water Extract of Liriope platyphylla on the mRNA Expression and Protein Secretion of Nerve Growth Factors (맥문동 유수추출물이 NGF의 mRNA발현과 단백질 분비에 미치는 영향)

  • Choi, Sun-Il;Park, Jee-Hee;Her, Youn-Kyng;Lee, Yoen-Kyung;Kim, Ji-Eun;Nam, So-Hee;Goo, Jun-Seo;Jang, Min-Ju;Lee, Hee-Sup;Son, Hong-Ju;Lee, Chung-Yeol;Hwang, Dae-Youn
    • Korean Journal of Medicinal Crop Science
    • /
    • v.18 no.5
    • /
    • pp.291-297
    • /
    • 2010
  • Liriope platyphylla has been though as an useful medical plant to improve the cough, sputum, neurodegenerative disorders, obesity, and diabetes in Korea and China from old times. In order to investigate the effects of Liriope platyphylla on expression and secretion of nerve growth factor (NGF), the mRNA expression and protein secretion were detected in the neuronal cell (B35) and neuroglial cell (C6) cultured with three differences concentration (5%, 10%, 15%) of Liriope platyphylla. In MTT assay and FACS anslysis, the some death of some B35 and C6 cells were observed in 15% extract-treated group, while other groups did not induce the death. Also, the mRNA expression of NGF were significantly increased in 5% and 10% extracts treated-group. Furthermore, the NGF protein concentration in supernatant collected from cultured cells showed the very similar pattern with mRNA expression. In order to verify the activity of secreted NGF, the culture supernatant collected from B35 and C6 cells cultured with Liriope platyphylla extracts for 24 hrs were treated into undifferentiated PC12 cells, and the differentiation level of PC12 cell were also observed with microscopes. The differentiation level of PC12 cell were significantly increased depend on the dose of extract. Therefore, these results suggested that the water extracts of Liriope platyphylla may contribute the regulation of NGF expression and secretion in the neuronal cell and be considered as an excellent candidate for a neurodegenerative disease-therapeutic drug.

Anti-proliferative Activity of Ethanol Extracts of Root of Aralia cordata var. continentalis through Proteasomal Degradation of Cyclin D1 in Human Colorectal Cancer Cells (독활 에탄올 추출물의 대장암 세포에서 Cyclin D1 단백질 분해 유도를 통한 세포 생육 억제활성)

  • Park, Su Bin;Park, Gwang Hun;Song, Hun Min;Park, Ji Hye;Shin, Myeong Su;Son, Ho Jun;Um, Yurry;Jeong, Jin Boo
    • Korean Journal of Medicinal Crop Science
    • /
    • v.25 no.5
    • /
    • pp.328-334
    • /
    • 2017
  • Background: In this study, we evaluated the anti-cancer activity and potential molecular mechanism of 70% ethanol extracts of the root of Aralia cordata var. continentalis (Kitagawa) Y. C. Chu (RAc-E70) against human colorectal cancer cells. Methods and Results: RAc-E70 suppressed the proliferation of the human colorectal cancer cell lines, HCT116 and SW480. Although RAc-E70 reduction cyclin D1 expression at the protein and mRNA levels, RAc-E70-induced reduction in cyclin D1 protein level occurred more dramatically than that of cyclin D1 mRNA. The RAc-E70-induced downregulation of cyclin D1 expression was attenuated in the presence of MG132. Additionally, RAc-E70 reduced HA-cyclin D1 levels in HCT116 cells transfected with HA-tagged wild type-cyclin D1 expression vector. RAc-E70-mediated cyclin D1 degradation was blocked in the presence of LiCl, a $GSK3{\beta}$ inhibitorbut, but not PD98059, an ERK1/2 inhibitor and SB203580, a p38 inhibitor. Furthermore, RAc-E70 phosphorylated cyclin D1 at threonine-286 (T286), and LiCl-induced $GSK3{\beta}$ inhibition reduced the RAc-E70-mediated phosphorylation of cyclin D1 at T286. Conclusions: Our results suggested that RAc-E70 may downregulate cyclin D1 expression as a potential anti-cancer target through $GSK3{\beta}$-dependent cyclin D1 degradation. Based on these findings, RAc-E70 maybe a potential candidate for the development of chemopreventive or therapeutic agents for human colorectal cancer.

Cytotoxicity of a cysteine proteinase of adult Clonorchis sinensis (간흡충의 cysteine 단백분해효소의 세포독성)

  • Hyun PARK;Man Young KO;Moon Kee PAIK;Ching Thack SOH;Jang Hoon SEO;Kyung-il IM
    • Parasites, Hosts and Diseases
    • /
    • v.33 no.3
    • /
    • pp.211-218
    • /
    • 1995
  • To clarify the correlation of the proteinase activity with pathogenicity of Clonorrhis sinensis, the proteinase activity either in excretory-secretory products (ESP) or in crude extracts of adult C. sinensis was examined. Substrate gel electrophoresis of the ESP and crude extracts revealed four distinct enzyme bands, which were differently inhibited by the specific proteinase inhibitors. The proteinase of the ESP with molecular mass of 24 kDa, was purified 23-fold with 14.5% yield by spectra gel ACA 44 gel filtration. It exhibited optimal pH at 7.5 in sodium phosphate (0.1 M). Its activity was inhibited specifically by N-ethylmaleimide (NEMI and antipain whereas potentiated 1.9 folds in the presence of 5 mM dithiothreitol (DTT). Cytotoxicity of the proteinase increased in a dose- dependent manner up to 120 ㎍/ml while reduced by NEM and antipain, indicating that cysteine proteinase was responsible for the cytotoxicity. This result shows that the 24 kDa cysteine proteinase is deeply correlated with the pathogenicity of C. sinensis infection.

  • PDF

Effects of dietary polyphenol (-)-epigallocatechin-3-gallate on the differentiation of mouse C2C12 myoblasts (식이성 폴리페놀 (-)-epigallocatechin-3-gallate가 mouse C2C12 myoblast 분화에 미치는 영향)

  • Kim, Hye-Jin;Lee, Won-Jun
    • Journal of Life Science
    • /
    • v.17 no.3 s.83
    • /
    • pp.420-426
    • /
    • 2007
  • In the present investigation, we studied the modulating effects of (-)-epigallocatechin-3-gallate(EGCG) on the differentiation of mouse C2C12 myoblasts. We found that the strong inhibitory effect of EGCG on DNA methyltransferase-mediated DNA methylation induced transdifferentiation of C2C12 myoblasts into smooth muscle cells demonstrated by both morphological changes and immunofluorescent staining. C2C12 myoblasts treated with EGCG for 4 days expressed smooth muscle ${\alpha}-actin$ protein. Real-time PCR data revealed that smooth muscle ${\alpha}-actin$ mRNA was induced by EGCG treated C2C12 myoblasts in a concentration-dependent manner. Smooth muscle ${\alpha}-actin$ mRNA concentration increased 330% and 490% after 2 and 3 days of 50 ${\mu}M$ of EGCG treatment. The expression of another smooth muscle marker, transgelin, mRNA was also increased up to 9-fold by 4 days of EGCG treatment compared with control in a concentration-dependent manner. These results suggested that C2C12 enables to transdifferentiate into smooth muscle when gene expression patterns are changed by the inhibition of DNA methylation induced by EGCG. In conclusion, transdifferentiation of C2C12 myoblasts into smooth muscle is resulted from the modulating effects of EGCG on DNA methylation which subsequently results in changing the expression pattern of several genes playing a critical role in the differentiation of C2C12 myoblasts.

Knowledge based Genetic Algorithm for the Prediction of Peptides binding to HLA alleles common in Koreans (지식기반 유전자알고리즘을 이용한 한국인 빈발 HLA 대립유전자에 대한 결합 펩타이드 예측)

  • Cho, Yeon-Jin;Oh, Heung-Bum;Kim, Hyeon-Cheol
    • Journal of Internet Computing and Services
    • /
    • v.13 no.4
    • /
    • pp.45-52
    • /
    • 2012
  • T cells induce immune responses and thereby eliminate infected micro-organisms when peptides from the microbial proteins are bound to HLAs in the host cell surfaces, It is known that the more stable the binding of peptide to HLA is, the stronger the T cell response gets to remove more effectively the source of infection. Accordingly, if peptides (HLA binder) which can be bound stably to a certain HLA are found, those peptieds are utilized to the development of peptide vaccine to prevent infectious diseases or even to cancer. However, HLA is highly polymorphic so that HLA has a large number of alleles with some frequencies even in one population. Therefore, it is very inefficient to find the peptides stably bound to a number of HLAs by testing random possible peptides for all the various alleles frequent in the population. In order to solve this problem, computational methods have recently been developed to predict peptides which are stably bound to a certain HLA. These methods could markedly decrease the number of candidate peptides to be examined by biological experiments. Accordingly, this paper not only introduces a method of machine learning to predict peptides binding to an HLA, but also suggests a new prediction model so called 'knowledge-based genetic algorithm' that has never been tried for HLA binding peptide prediction. Although based on genetic algorithm (GA). it showed more enhanced performance than GA by incorporating expert knowledge in the process of the algorithm. Furthermore, it could extract rules predicting the binding peptide of the HLA alleles common in Koreans.

Helixor A Inhibits Angiogenesis in vitro Via Upregutation of Thrombospondin-1 (Helixor A는 시험관 내에서 thrombospondin-1의 상승조절을 통해 신혈관생성을 억제한다.)

  • Yeom Dong-Hoon;Hong Kyong-Ja
    • Journal of Life Science
    • /
    • v.15 no.6 s.73
    • /
    • pp.895-903
    • /
    • 2005
  • Thrombospondin-1 (TSP-1), a negative regulator in tumor growth and angiogenesis, is cell-type specifically regulated under pathological conditions or by extracellular stimuli, and the regulation of TSP-1 gene expression is important for developing new approaches in tumor therapy. Mistletoe is a parasitir plant that have been used for immunomodulation and antitumor therapy. Helixor A is an aqueous part of mistletoes extract. Here we showed that TSP-1 expression was significantly induced at both mRNA and protein levels in the Hepatocarcinorna cell line (Hep3B) and primary bovine endothelial cell line (BAE) exposed to Helixor A. Our promoter analysis confirmed that the expression of TSP-1 gene was regulated by Helixor A at the transcriptional level. In cell invasion assay, the conditioned media obtained from treatment of these cells significantly reduced the number of invasive cells and also inhibited capillary-like tube formation of BAE cells on Matrigel. Moreover, the inhibitory efforts of the conditioned media on cell invasion and tube formation were reversed by blocking with anti-TSP-1 neutralizing antibodies, suggesting that TSP-1 is involved in Helixor A-indured antiangiogenic effect. Taken together, our results suggest that Helixor A have an antiangiogenic effects through upregulation of TSP-1.

Effects of Propolis Addition on Quality Characteristics of Oriental Medicinal Seasoning Pork (Propolis 첨가가 한방양념돼지고기의 품질특성에 미치는 영향)

  • Han, Gwi-Jeong;Shin, Dong-Sun;Kim, Jin-Sook;Cho, Yong-Sik;Jeong, Kyoung-Soon
    • Korean Journal of Food Science and Technology
    • /
    • v.38 no.1
    • /
    • pp.75-81
    • /
    • 2006
  • Effects of medicinal herbs and bee propolis on pork quality were assessed using formulae A (control), B (boiling water extraction with propolis), and C (boiling water extraction without propolis). Seasoning sauce was made with medicinal herb, spices, total flavonoid, and soy sauce with/without 0.4% propolis (21.8 mg/g), mixed with pork, and stored at low temperature $(4{\pm}1^{\circ}C)$ for 20 days. In formula B peroxide value and thiobarbituric acid content increased with increasing storage period. Formula B showed highest Volatile Basic Nitrogen (VBN) and lowest microbial counts, followed by formulae C and A, receiving higher scores for taste and fragrance.

The VHR Dual-Specificity Protein Tyrosine Phosphatase (DS-PTPase) Inhibitor Isolated from Gastrodia elata (천마(Gastrodia elata)로부터 분리한 VHR DS-PTPase 저해 물질)

  • Lee, Myung-Sun;Oh, Won-Keun;Bae, Eun-Young;Ahn, Soon-Cheol;Sohn, Cheon-Bae;Osada, Hiroyuki;Ahn, Jong-Seog
    • Korean Journal of Food Science and Technology
    • /
    • v.34 no.3
    • /
    • pp.505-509
    • /
    • 2002
  • In the course of screening program for VHR DS-PTPase (dual-specificity protein tyrosine phosphatase) from natural sources, Gastrodia elata was selected. One compound showing potent inhibitory activity was isolated by the solvent extraction and column chromatography including silica gel, ODS RP-18, Sephades LH-20, and HPLC. This compound was identified as baicalein by several NMR techniques such as $^1H-NMR$, $^{13}C-NMR$, and DEPT. Baicalein showed selective inhibitory activity against VHR DS-PTPase with $IC_{50}=2.4\;{\mu}M$, and showed cytotoxicity against several human cancer cell lines with an $GI_{50}$ of $5.26{\sim}12.93\;{\mu}g/mL$ range, including, melanoma (LOX-IMVI), lung cancer (NCI H23 and A549), colon cancer (HCT 116 and SW 620), prostate cancer (PC-3), and leukemia (MOLT 4F).

Mining Maximal Frequent Contiguous Sequences in Biological Data Sequences (생물학적 데이터 서열들에서 빈번한 최대길이 연속 서열 마이닝)

  • Kang, Tae-Ho;Yoo, Jae-Soo
    • The KIPS Transactions:PartD
    • /
    • v.15D no.2
    • /
    • pp.155-162
    • /
    • 2008
  • Biological sequences such as DNA sequences and amino acid sequences typically contain a large number of items. They have contiguous sequences that ordinarily consist of hundreds of frequent items. In biological sequences analysis(BSA), a frequent contiguous sequence search is one of the most important operations. Many studies have been done for mining sequential patterns efficiently. Most of the existing methods for mining sequential patterns are based on the Apriori algorithm. In particular, the prefixSpan algorithm is one of the most efficient sequential pattern mining schemes based on the Apriori algorithm. However, since the algorithm expands the sequential patterns from frequent patterns with length-1, it is not suitable for biological dataset with long frequent contiguous sequences. In recent years, the MacosVSpan algorithm was proposed based on the idea of the prefixSpan algorithm to significantly reduce its recursive process. However, the algorithm is still inefficient for mining frequent contiguous sequences from long biological data sequences. In this paper, we propose an efficient method to mine maximal frequent contiguous sequences in large biological data sequences by constructing the spanning tree with the fixed length. To verify the superiority of the proposed method, we perform experiments in various environments. As the result, the experiments show that the proposed method is much more efficient than MacosVSpan in terms of retrieval performance.