• Title/Summary/Keyword: affinity matrix

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Biocompatibility and Bone Conductivity of Porous Calcium Metaphosphate Blocks (생분해성 다공질 Calcium Metaphosphate 블록의 조직적합성에 관한 연구)

  • Lee, Yong-Moo;Kim, Seok-Young;Shin, Seung-Yun;Ku, Young;Rhyu, In-Chul;Chung, Chong-Pyoung
    • Journal of Periodontal and Implant Science
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    • v.28 no.4
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    • pp.559-568
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    • 1998
  • direct bone apposition during bone remodelling. To address these problem, we developed a new ceramic, calcium metaphosphate(CMP), and report herein the biologic response to CMP in subcutaneous tissue, muscle and bone. Porous CMP blocks were prepared by condensation of anhydrous $Ca(H_2PO_4)_2$ to form non-crystalline $Ca(PO_3)_2$. Macroporous scaffolds were made using a polyurethane sponge method. CMP block possesses a macroporous structure with approximate pore size range of 0.3-1mm. CMP blocks were implanted in 8mm sized calvarial defect, subcutaneous tissue and muscle of 6 Newzealand White rabbits and histologic observation were performed at 4 and 6 weeks later. CMP blocks in subcutaneous tissue and muscle were well adapted without any adverse tissue reaction and resorbed slowly and spontaneously. Histologic observation of calvarial defect at 4 and 6 weeks revealed that CMP matrix were mingled with and directly apposed to new bone without any intervention of fibrous connective tissue. CMP blocks didn't show any adverse tissue reaction and resorbed spontaneously also in calvarial defect. This result revealed that CMP had a high affinity for bone and was very biocompatible. From this preliminary result, it was suggested that CMP was a promising ceramic as a bone substitute and tissue engineering scaffold for bone formation.

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Superoxide Dismutase Isoenzyme Activities in Plasma and Tissues of Iraqi Patients with Breast Cancer

  • Hasan, Hathama Razooki;Mathkor, Thikra Hasan;Al-Habal, Mohammed Hasan
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.6
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    • pp.2571-2576
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    • 2012
  • Breast cancer is the first of the most common ten cancers in Iraq. Its etiology is multifactorial, oxidative stress and lipid peroxidation being suggested to play important roles in carcinogenesis. The purpose of this study was to investigate the oxidant-antioxidant status in breast cancer patients, by measuring SOD isoenzyme activities (total SOD, CuZn-SOD, Mn-SOD and EC-SOD) in plasma and breast tumors, and by estimating thiobarbituric reactive substances (TBRS) in tissue homogenates. General increase in total SOD activity was observed in plasma and tissue samples of breast tumors, greater in the malignant when compared to benign group (p<0.05). Mn-SOD showed a significant decrease in tissue malignant samples (p<0.05), and insignificant decrease in plasma malignant samples compared with control and benign samples. Plasma EC-SOD activity in both patient benign and malignant breast tumors demonstrated 3.5% and 22.8% increase, respectively. However, there was a decrease in tissue EC-SOD activity in malignant breast tumors when compared with benign. A similar tendency was noted for TBRS. We suggest that elevated total SOD might reflect a response to oxidative stress, and then may predict a state of excess reactive oxygen species in the carcinogenesis process. If there is proteolytic removal of the heparin binding domain, EC-SOD will lose its affinity for the extracellular matrix and diffuse out of the tissue. This will result in a decreased EC-SOD activity, thus leading to an increase in the steady-state concentration of $O^{2-}$ in this domain, and increase in EC-SOD activity in the extracellular fluid. This might explain the results recorded here concerning the decrease in tissue EC-SOD activity and increase in plasma of breast cancer patients.

Proposing the Methods for Accelerating Computational Time of Large-Scale Commute Time Embedding (대용량 컴뮤트 타임 임베딩을 위한 연산 속도 개선 방식 제안)

  • Hahn, Hee-Il
    • Journal of the Institute of Electronics and Information Engineers
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    • v.52 no.2
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    • pp.162-170
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    • 2015
  • Commute time embedding involves computing the spectral decomposition of the graph Laplacian. It requires the computational burden proportional to $o(n^3)$, not suitable for large scale dataset. Many methods have been proposed to accelerate the computational time, which usually employ the Nystr${\ddot{o}}$m methods to approximate the spectral decomposition of the reduced graph Laplacian. They suffer from the lost of information by dint of sampling process. This paper proposes to reduce the errors by approximating the spectral decomposition of the graph Laplacian using that of the affinity matrix. However, this can not be applied as the data size increases, because it also requires spectral decomposition. Another method called approximate commute time embedding is implemented, which does not require spectral decomposition. The performance of the proposed algorithms is analyzed by computing the commute time on the patch graph.

Surface Polarity Dependent Solid-state Molecular Biological Manipulation with Immobilized DNA on a Gold Surface

  • Lee, Jiyoung;Kim, Jeong Hee
    • International Journal of Oral Biology
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    • v.37 no.4
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    • pp.181-188
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    • 2012
  • As the demand for large-scale analysis of gene expression using DNA arrays increases, the importance of the surface characterization of DNA arrays has emerged. We compared the efficiency of molecular biological applications on solid-phases with different surface polarities to identify the most optimal conditions. We employed thiol-gold reactions for DNA immobilization on solid surfaces. The surface polarity was controlled by creating a self-assembled monolayer (SAM) of mercaptohexanol or hepthanethiol, which create hydrophilic or hydrophobic surface properties, respectively. A hydrophilic environment was found to be much more favorable to solid-phase molecular biological manipulations. A SAM of mercaptoethanol had the highest affinity to DNA molecules in our experimetns and it showed greater efficiency in terms of DNA hybridization and polymerization. The optimal DNA concentration for immobilization was found to be 0.5 ${\mu}M$. The optimal reaction time for both thiolated DNA and matrix molecules was 10 min and for the polymerase reaction time was 150 min. Under these optimized conditions, molecular biology techniques including DNA hybridization, ligation, polymerization, PCR and multiplex PCR were shown to be feasible in solid-state conditions. We demonstrated from our present analysis the importance of surface polarity in solid-phase molecular biological applications. A hydrophilic SAM generated a far more favorable environment than hydrophobic SAM for solid-state molecular techniques. Our findings suggest that the conditions and methods identified here could be used for DNA-DNA hybridization applications such as DNA chips and for the further development of solid-phase genetic engineering applications that involve DNA-enzyme interactions.

Tissue Inhibitor of Metalloproteinases-2 Inhibits the 4-Aminophenylmercuric Acetate-Induced Activation and Autodegradation of the Free Promatrix Metalloproteinase-2

  • Jo, Yi-Hyung;Yoon, Dae-Woong;Kim, Min-Young;Lee, Yoon-Ju;Kim, Hwa-Jung;Lee, Seung-Taek
    • BMB Reports
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    • v.32 no.1
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    • pp.60-66
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    • 1999
  • Matrix metalloproteinase-2 (MMP-2; 72-kDa gelatinase; 72-kDa type IV collagenase; gelatinase A) plays an important role in normal physiological processes and in many pathologic processes such as arthritis and metastasis of cancer. Tissue inhibitor of metalloproteinases-2 (TIMP-2) binds to proMMP-2 or mature MMP-2 at a 1:1 ratio and inhibits the catalytic activity of MMP-2. We demonstrated that the baculovirus/insect cell system does not have TIMP-2 activity. The human proMMP-2 free of TIMP-2 was expressed in the expression system and purified by one-step affinity chromatography using gelatin-Sepharose. The free proMMP-2 was autoactivated to the mature MMP-2 and autodegraded into smaller molecular weight forms in the absence of external activator. The activation and autodegradation of the proMMP-2 was much more rapid in the presence of 4-aminophenylmercuric acetate (APMA). Addition of TIMP-2 inhibits both APMA-induced activation and autodegradation of the free proMMP-2. However, an increasing concentration of TIMP-2 more readily inhibited activation of the free proMMP-2 than autodegradation. These results demonstrate that TIMP-2 plays roles in inhibition of both activation and autodegradation of the free proMMP-2 in addition to inhibition of the catalytic activity of MMP-2.

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Characteristics of IEF Patterns and SDS-PAGE Results of Korean EPO Biosimilars

  • Kang, Min-Jung;Shin, Sang-Mi;Yoo, Hey-Hyun;Kwon, Oh-Seung;Jin, Chang-Bae
    • Bulletin of the Korean Chemical Society
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    • v.31 no.9
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    • pp.2493-2496
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    • 2010
  • Erythropoietin (EPO) is mainly produced in kidney and stimulates erythropoiesis. The use of recombinant EPOs for doping is prohibited because of its performance enhancing effect. This study investigated whether biosimilar EPOs could be differentiated from endogenous one by iso-electro-focusing plus double blotting and SDS-PAGE for antidoping analysis. The established method was validated with positive control urine. The band patterns were reproducible and meet the criteria, which was made by world anti doping agency (WADA). Isoelectric focusing was conducted in pH range 2 to 6. Recormon (La Roche), Aropotin (Kunwha), Epokine (CJ Pharm Co.), Eporon (Dong-A), Espogen (LG Life Sciences), and Dynepo (Shire Pharmaceuticals) were detected in basic region. All biosimilars showed discriminative isoelectric profiles from endogenous EPO profiles, but they showed different band patterns with the reference one except Epokine (CJ Pharm Co.). Next, SDS-PAGE of biosimilar EPOs resulted in different molecular weight patterns which were distributed higher than endogenous EPO. Commercial immune assay kit as an immune affinity purification tool and immobilized antibody coated magnetic bead were tested for the purification and concentration of EPO from urinary matrix. The antibody-coated magnetic bead gave better purification yield. The IEF plus double blotting and SDS-PAGE with immunoaffinity purification method established can be used to discriminate biosimilar EPOs from endogenous EPO.

Bio-Composite Materials Precursor to Chitosan in the Development of Electrochemical Sensors: A Critical Overview of Its use with Micro-Pollutants and Heavy Metals Detection

  • Sarikokba, Sarikokba;Tiwari, Diwakar;Prasad, Shailesh Kumar;Kim, Dong Jin;Choi, Suk Soon;Lee, Seung-Mok
    • Applied Chemistry for Engineering
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    • v.31 no.3
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    • pp.237-257
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    • 2020
  • The role of nano bio-composites precursor to chitosan are innumerable and are known for having different applications in various branches of physical sciences. The application to the sensor development is relatively new, where only few literature works are available to address the specific and critical analysis of nanocomposites in the subject area. The bio-composites are potential and having greater affinity towards the heavy metals and several micro-pollutants hence, perhaps are having wider implications in the low or even trace level detection of the pollutants. The nano-composites could show good selectivity and suitability for the detection of the pollutants as they are found in the complex matrix. However, the greater challenges are associated using the bio-composites, since the biomaterials are prone to be oxidized or reduced at an applied potential and found to be a hinderance for the detection of target pollutants. In addition, the materials could proceed with a series of electrochemical reactions, which could produce different by-products in analytical applications, resulting in several complex phenomena in electrochemical processes. Therefore, this review addresses critically various aspects of an evaluation of nano bio-composite materials in the electrochemical detection of heavy metals and micro-pollutants from aqueous solutions.

Determination of Optimum Bead Size by Calculating Effectiveness Factors in Cyclosporin A Fermentation by Immobilized Cells (Cyclosporin A 고정상 발효에서 효율인자의 계산을 통한 고정화 담체의 최적크기 결정)

  • 전계택;이태호장용근
    • KSBB Journal
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    • v.11 no.1
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    • pp.30-36
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    • 1996
  • Based on fermentation data for cyclosporin A production, simple Monod kinetics was proposed for both immobilized and suspended cultures. Higher value of $\mu$mas and lower value of Km suggest better catalytic activity of the immobilized cells than the parallel suspended cells. Furthermore, lower Km value in the immobilized cell system indicates higher affinity of the immobilized cells for carbon substrate as compared with the suspended cells. For immobilized cell cultures, these parameters were also utilized for the estimation of effectiveness factor, an indicator for intraparticle mass transfer resistance. Based on simulation studies, optimum radius of celite beads was turned out $100 ~ 500{\mu}m$In this simulation work, we examined the influence of biosupport size and immobilized biomass density on diffusional resistance of substrate inside the bead matrix. In order to maintain uniformly distributed cell activities in biosupport, it was essential to determine optimum slze of particle, and then to estimate the most economic loaded biomass content.

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Production and Characterization of Monoclonal and Recombinant Antibodies Against Antimicrobial Sulfamethazine

  • Yang, Zheng-You;Shim, Won-Bo;Kim, Min-Gon;Lee, Kyu-Ho;Kim, Keun-Sung;Kim, Kwang-Yup;Kim, Cheol-Ho;Ha, Sang-Do;Chung, Duck-Hwa
    • Journal of Microbiology and Biotechnology
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    • v.17 no.4
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    • pp.571-578
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    • 2007
  • A monoclonal antibody (mab) against the antimicrobial sulfamethazine was prepared and characterized by an indirect competitive enzyme-linked immunosorbent assay (IC-ELISA). Sulfamethazine in the range of 0.2 and 45ng/ml could be determined with the mab by IC-ELISA. cDNAs encoding a variable heavy chain and variable light chain of the mab were cloned to produce recombinant antibodies using phage display technology. Following phage rescue and three rounds of panning, a single-chain variable fragment (scFv) antibody with high sulfamethazine-binding affinity was obtained. ELISA analysis revealed that scFv antibody and parent mab showed similar, but not identical, characteristics. The $IC_{50}$ value by IC-ELISA with scFv antibody was 4.8ng/ml, compared with 1.6ng/ml with the parent mab. Performances of the assays in the presence of milk matrix were compared; the mab-based assay was less affected than the scFv-based assay. Sixty milk samples were analyzed by mab-based IC-ELISA, and four samples were sulfamethazine positive; these results were favorably correlated with those obtained by HPLC.

Preparation and Analysis of Yeast Cell Wall Mannoproteins, Immune Enhancing Materials, from Cell Wall Mutant Saccharomyces cerevisiae

  • Ha Chang-Hoon;Yun Cheol-Won;Paik Hyun-Dong;Kim Seung-Wook;Kang Chang-Won;Hwang Han-Joon;Chang Hyo-Ihl
    • Journal of Microbiology and Biotechnology
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    • v.16 no.2
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    • pp.247-255
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    • 2006
  • Yeast cell wall matrix particles are composed entirely of mannoprotein and ${\beta}-glucan$. The mannoproteins of yeast cell wall can systemically enhance the immune system. We previously purified and analyzed alkali-soluble ${\beta}-glucans$ [${\beta}$-(1,3)- and ${\beta}$-(1,6)-glucans] [10]. In the present study, a wild-type strain was first mutagenized with ultraviolet light, and the cell wall mutants were then selected by treatment with 1.0 mg/ml laminarinase (endo-${\beta}$-(1,3)-D-glucanase). Mannoproteins of Saccharomyces cerevisiae were released by laminarinase, purified by concanavalin-A affinity and ion-exchange chromatography. The results indicated that the mutants yielded 3-fold more mannoprotein than the wild-type. The mannoprotein mass of mutant K48L3 was 2.25 mg/100 mg of yeast cell dry mass. Carbohydrate analysis revealed that they contained mannose, glucose, and N-acetylglucosamine. Saccharomyces cerevisiae cell wall components, mannoproteins, are known to interact with macrophages through receptors, thereby inducing release of tumor necrosis factor alpha ($TNF-{\alpha}$) and nitric oxide. Mannoprotein tractions in the present study had a higher macrophage activity of secretion of $TNF-{\alpha}$ and nitric oxide and direct phagocytosis than positive control ($1{\mu}g$ of lipopolysaccharide). In particular, F1 and F3 fractions in mannoproteins of K48L3 enhanced and upregulated the activity of nitric oxide secretion and macrophage phagocytosis by approximately two- and four-fold, respectively.