• Title/Summary/Keyword: proteoglycans

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Purification of Mucilages from Dioscorea batatas and D.japonica and their Content Analysis (산약(山藥) 점액성분(粘液成分)의 정제(精製)와 함량분석(含量分析)에 관한 연구(硏究))

  • Han, Yong-Nam;Hahn, Seung-Hye;Lee, Ihn-Rhan
    • Korean Journal of Pharmacognosy
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    • v.21 no.4
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    • pp.274-283
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    • 1990
  • This study was conducted to further characterize mucilages from Dioscoreae Rhizoma, which have been known to be proteoglycans. We chose the two types of yams, Dioscorea batatas Dec. (club-like) and D. japonica Thunb. (cane-like). Repeated gel filtration of a dialyzed water extract of each fresh yam on Bio-gel P-100 and then Sephadex G-150 columns completely separated the mucilage from protein. Furthermore, gel filtration of a water extract from each yam processed by steaming and drying on the Bio-gel P-100 column gave only one polysaccharide peak without protein. These results revealed that the mucilages of yams were only composed of polysaccharide. Then we assessed some properties of the mucilages under three kinds of criteria: a complex-forming capacity between mucilage and alcian blue, mannose content in the mucilage, and viscosity. The complex-forming capacities of two types of fresh yams were closely similar with each other, but the processing of two types of the fresh yams greatly lowered the complex-forming capacity and viscosity.

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Characterization of the Pathogenesis Mechanism after Pseudomonas aeruginosa Infection through Food Consumption Using Chick Embryo Model

  • Song, Jin-Soo;Jin, Eun-Jung;Choi, Kyoung-Hee
    • Food Science of Animal Resources
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    • v.30 no.4
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    • pp.568-574
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    • 2010
  • This study introduced a chick embryos’ infection model to elucidate the pathogenesis mechanism of Pseudomonas aeruginosa, which causes serious diseases in human after ingestion of P. aeruginosa-contaminated animal originated foods. The embryonic chick model is able to give a rapid and relatively inexpensive method to assess bacterial pathogenicity compared to embryos of other vertebrates. Embryos were infected with P. aeruginosa and elastase-deficient P. aeruginosa. After infection with P. aeruginosa cells, total bacterial cell numbers and gelatinase activities in the embryos were compared. Thereafter, precartilage condensation and chondrogenesis were assessed by peanut agglutinin (PNA) binding on day 3 and by Alcian blue staining for sulfated proteoglycans on day 5, respectively. P. aeruginosa significantly increased in embryos, resulting in abnormal limb development, whereas P. aeruginosa defective in elastase activity partly impaired proliferation. In addition, P. aeruginosa-infected chick embryos significantly stimulated the production of matrix metalloproteinases. Several analyses showed that elevated proteases suppressed the proliferation and survival of chondrogenic cells. The results show that this infection model was a useful assay to determine the virulence mechanism of P. aeruginosa in human after intake of microbiologically contaminated foods.

Inhibitory Effect of Sparassis crispa (Wulf.) Extract on Monosodium Iodoacetate Induced Osteoarthritis (꽃송이버섯 추출물의 Monosodium Iodoacetate로 유도된 골관절염 억제 효과)

  • Kim, Eun-Nam;Roh, Seong-Soo;Jeong, Gil-Saeng
    • Korean Journal of Pharmacognosy
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    • v.49 no.3
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    • pp.262-269
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    • 2018
  • Sparassis crispa (Wulf.) is an edible/medicinal mushroom and has been reported to biological activities such as antitumor, anti-angiogenesis, antioxidant and wound healing. However, there have not been many researches on osteoarthritis of S. crispa. The aim of this study was to investigate the effects of S. crispa extract on rats with osteoarthritis induced by MIA. Osteoarthritis is a gradually developmental disease that early stage, causes joint stiffness and complains of joint pain. In addition, it gives rise to edema and hypo-function. The results of this study, S. crispa extract effectively inhibited ROS production, increased the production of antioxidant protein SOD and catalase in knee joint cartilage tissue. In addition, S. crispa extract inhibited the expression of pro-inflammatory cytokines and enzymes such as NOX4 and $P47^{phox}$, which are involved in the expression of COX-2, iNOS and the production of ROS. Also, S. crispa extract inhibited the destruction of synovial tissue, cartilage tissue and proteoglycans in articular cartilage in rats.

Effect of Achyrantis Radixs and Ultrasound in Osteoarthritis Rats Articular Cartilage (우슬과 초음파가 퇴행성관절염 백서의 관절 연골에 미치는 영향)

  • Kim, Eun-Jung;Chung, Hun-Woo;Kim, Gye-Yeop
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.22 no.2
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    • pp.390-396
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    • 2008
  • Osteoarthritis(OA) diseases are characterized by joint pain, tenderness, limitation of movement, crepitus, occasional effusion, and variable degrees of inflammation without systemic effects. We investigated the effects of Achyrantis radixs cream treatment and low intensity ultrasound in monosodium iodoacetate(MIA) induced experimental osteoarthritis rat. Sprague-Dawley 40 rats of 7-8 weeks, weight $250\;{\pm}\;50$ g were divided into four groups including the control group and ostoarthritis group(30 rats). Histopathological examination, Mankin's score, and immunohistochemical were performed. Histological findings in control group that are similar to those observed in human osteoarthritis, such as disorganization of chondrocytes, erosion and fibrillation of cartilage surface, and subchondral bone exposure. Safranin O-fast green staining revealed that marked diffuse reduction of proteoglycans and chondrocyte treated with MIA. The Mankin's score were closely correlated to the grade of histological findings. The level of Bax and caspase-3 expression decreased experimental groups. This study shows that a Acyranthes Radix cream treatment and low intensity ultrasound exerts a beneficial influence on the severity of chondral lesion in osteoarthritis rats. This treatments could related to a reduced level of chondrocyte apoptosis through anti-apoptotoc capacities of MIA-induced apoptotic protein overexpression.

High-yield Expression and Characterization of Syndecan-4 Extracellular, Transmembrane and Cytoplasmic Domains

  • Choi, Sung-Sub;Kim, Ji-Sun;Song, Jooyoung;Kim, Yongae
    • Bulletin of the Korean Chemical Society
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    • v.34 no.4
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    • pp.1120-1126
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    • 2013
  • The syndecan family consists of four transmembrane heparan sulfate proteoglycans present in most cell types and each syndecan shares a common structure containing a heparan sulfate modified extracellular domain, a single transmembrane domain and a C-terminal cytoplasmic domain. To get a better understanding of the mechanism and function of syndecan-4 which is one of the syndecan family, it is crucial to investigate its three-dimensional structure. Unfortunately, it is difficult to prepare the peptide because it is membrane-bound protein that transverses the lipid bilayer of the cell membrane. Here, we optimize the expression, purification, and characterization of transmembrane, cytoplasmic and short extracellular domains of syndecan4 (syndecan-4 eTC). Syndecan-4 eTC was successfully obtained with high purity and yield from the M9 medium. The structural information of syndecan-4 eTC was investigated by MALDI-TOF mass (MS) spectrometry, circular dichroism (CD) spectroscopy, and nuclear magnetic resonance (NMR) spectroscopy. It was confirmed that syndecan-4 eTC had an ${\alpha}$-helical multimeric structure like transmembrane domain of syndecan-4 (syndecan-4 TM) in membrane environments.

Regeneration of a Cartilage Tissue by In Vitro Culture of Chondrocytes on PLGA Microspheres

  • Son, Jeong-Hwa;Park, So-Ra;Kim, Hyeon-Joo;Min, Byoung-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.16 no.10
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    • pp.1577-1582
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    • 2006
  • Cartilage tissue engineering has emerged as an alternative approach for reconstruction or repair of injured cartilage tissues. In this study, rabbit chondrocytes were cultured in a three-dimensional environment to fabricate a new cartilaginous tissue with the application of tissue engineering strategies based on biodegradable PLGA microspheres. Chondrocytes were seeded on PLGA microspheres and cultured on a rocking platform for 5 weeks. The PLGA microspheres provided more surface area to adhere chondrocytes compared with PLGA sponge scaffolds. The novel system facilitated uniform distribution of the cells on the whole of the PLGA microspheres, thus forming a new cartilaginous construct at 4 weeks of culture. The histological and immunohistochemical analyses verified that the number of chondrocytes and the amount of extracellular matrix components such as proteoglycans and type II collagen were significantly greater on the PLGA microspheres constructs as compared with those on the PLGA sponge scaffolds. Therefore, PLGA microspheres enhanced the function of chondrocytes compared with PLGA sponge scaffolds, and thus might be useful for formation of cartilage tissue in vitro.

NMR Structure of Syndecan-4L reveals structural requirement for PKC signalling

  • Koo, Bon-Kyoung;Joon Shin;Oh, Eok-Soo;Lee, Weontae
    • Proceedings of the Korean Magnetic Resonance Society Conference
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    • 2002.08a
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    • pp.90-90
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    • 2002
  • Syndecans, transmembrane heparan sulfate proteoglycans, are coreceptors with integrin in cell adhesion process. It forms a ternary signaling complex with protein kinase C and phosphatidylinositol 4,5 bisphosphate (PIP2) for integrin signaling. NMR data indicates that cytoplasmic domain of syndecan-4 (4L) undergoes a conformational transition in the presence of PIP2, forming oligomeric conformation. The structure based on NMR data demonstrated that syndecan-4L itself forms a compact intertwined symmetric dimer with an unusual clamp shape for residues Leu$^{186}$ -Ala$^{195}$ . The molecular surface of the syndecan-4L dimer is highly positively charged. In addition, no inter-subunit NOEs in membrane proximal amino acid resides (Cl region) has been observed, demonstrating that the Cl region is mostly unstructured in syndecan-4L dimmer. However, the complex structure in the presence of PIP2 induced a high order multimeric conformation in solution. In addition, phosphorylation of cytoplasmic domain induces conformational change of syndecan-4, resulting inhibition of PKC signaling. The NMR structural data strongly suggest that PIP2 promotes oligomerization of syndecan-4 cytoplasmic domain for PKC activation and further induces structural reorganization of syndecan for mediating signaling network in cell adhesion procedure.

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Development of Extracellular Matrix (ECM) based Dermal Filler (세포외기질(ECM) 생체소재 기반 필러 개발 연구)

  • Kim, Na Hyeon;Park, Sang-Hyug
    • Journal of Biomedical Engineering Research
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    • v.40 no.4
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    • pp.137-142
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    • 2019
  • Numerous efforts are being made to develop an ideal dermal filler that should be bio-compatibility, non-immunogenicity, long-lasting and biodegradable without a toxic secretion. Biomaterials of dermal fillers are hyaluronic acid filler, calcium filler, PMMA filler and collagen filler depending on the ingredient. Although hyaluronic acid (HA) is most widely used, it has shortages such as short shelf life and low mechanical strength compare to extracellular matrix (ECM). The cartilage ECM composed of collagen type II, proteoglycans, glycosaminoglycans (GAGs) and in a minor part with glycoproteins. In this study, we developed a cartilage ECM injectable filler capable of improving biocompatibility and longevity compared with hyaluronic acid (HA) fillers. The ECM hydrogel was cross-linked by the reaction of N-(3-Dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (EDC)/N-hydroxysuccinimide (NHS) for mechanical enhancement. Prepared ECM filler was compared with cross-linked HA by butanediol diglycidyle ether (BDDE), which is the most widely used natural polymers for dermal filler. In the results, the articular cartilage ECM hydrogel has great potential as a dermal filler to improve the biophysical and biological performance.

The effect of Gihyeolgwanjeolbang-A(CWS-A) on adjuvant-induced arthritis in rats (기혈관절방(氣血關節方)A(CWS-A)가 CFA로 유발(誘發)된 관절염(關節炎)에 미치는 영향(影響))

  • Na, Chang-Su;Youn, Dae-Hwan;Kim, Jeong-Sang;Chae, Woo-Seok
    • The Korea Journal of Herbology
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    • v.27 no.1
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    • pp.1-10
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    • 2012
  • Objective : To investigate effects of herbal preparations formulated(Gihyeolgwanjeolbang-A, CWS-A) on complete Freund's adjuvant (CFA)-induced arthritis in rats. Method : Arthritis was induced by injecting CFA subcutaneously into the left knee joint and paw, and the herbal preparations formulated(CWS-A I, 82.3 mg/kg ; CWS-A II, 164.6 mg/kg) was administered orally (i.g.) for 10 consecutive days beginning on day 10 after the injection. External shape, paw edema, inflammatory cytokines tumor necrosis factor alpha (TNF-alpha) and interleukin-1 beta (IL-1beta), and histological observation were assessed. Result : In swelling of the paw, CWS-A I and CWS-A II group in 15 days and 20 days were significantly reduced compared to controls. In serum ALT, CWS-A I and CWS-A II group were significantly reduced compared to controls. In TNF-${\alpha}$, CWS-A I and CWS-A II group were a tendency reduced compared to controls. In HGB, HCT, MCHC of erythrocytes, CWS-A II group was increased compared to controls. In histological observations, CWS-A II group was observed synoviocytes more than control group and was observed proteoglycans in the deep layers. Conclusion : The data suggest that CWS-A significant anti-arthritic effects that may be mediated by suppressing inflammatory parameters.

Cloning of a matrix metalloproteinase cDNA from Scylliorhinus torazame (두툽상어 matrix metalloproteinase 유전자 cDNA의 클로닝)

  • Kim, Jon Won;Cho, Won Jin;Chun, Kwang Ho;Kim, Kyu-Won;Kim, Yung-Jin;Lee, Sang-Jun;Shin, Hae-Ja;Lim, Woon Ki
    • Journal of Life Science
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    • v.8 no.3
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    • pp.235-240
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    • 1998
  • Matrix metalloproteinases(MMPs) are a group of zinc enzymes responsible for degradation of the matrix components such as collagen and proteoglycans in normal embryogenesis and remodeling and in many disease processes such as arthritis, cancer, periodontitis, and osteprocess. Genetically distince MMPs have been characterized and their genes have been cloned thus far from a variaty of species but not from fishes. In this stydy, a mmp cDNA was cloned by using RT-PCR(reverse transcriptase dependent polymerase chain reaction) from Scylliorhinus toraxzame(shark), agroup of cartilaginous fish, abundant in the coast of Pusan, Korea. It has 74% base homologue with membrane type matrix matalloproteinase-3 genes(mt3-mmps) from human, rat and chick, and also shows more than 90% residue homologue with them. In addition, it has cysteine switch domain, zinc binding domain(HExGH motif), propeptide cleavage site, and RRKR motif, which are present in MMPs. This result indicates that cDNA fragment cloned here may be mt3-mmp or its analogous gejne cDNA fragment of Scylliorhinus torzame.

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