• 제목/요약/키워드: Affinity Matrix

검색결과 96건 처리시간 0.022초

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

  • 이용무;김석영;신승윤;구영;류인철;정종평
    • Journal of Periodontal and Implant Science
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    • 제28권4호
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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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    • 제13권6호
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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.

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

  • 한희일
    • 전자공학회논문지
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    • 제52권2호
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    • pp.162-170
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    • 2015
  • 컴뮤트 타임 임베딩을 구현하려면 그래프 라플라시안 행렬의 고유값과 고유벡터를 구하여야 하는데, $o(n^3)$의 계산량이 요구되어 대용량 데이터에는 적용하기 어려운 문제가 있다. 이를 줄이기 위하여 표본화 과정을 통하여 크기가 줄어든 그래프 라플라시안 행렬에서 구한 다음, 원래의 고유값과 고유벡터를 근사화시키는 Nystr${\ddot{o}}$m 기법을 주로 채택한다. 이 과정에서 많은 오차가 발생하는데, 이를 개선하기 위하여 본 논문에서는 그래프 라플라시안 대신에 가중치 행렬을 표본화하고 이로부터 구한 고유값과 고유벡터를 그래프 라플라시안의 고유값과 고유벡터로 변환하는 기법을 이용하여 대용량 데이터로 구성된 스펙트럴 그래프를 근사적으로 컴뮤트 타임 임베딩하는 기법을 제안한다. 하지만, 이 방식도 스펙트럼 분해를 계산하여야 하므로 데이터의 크기가 증가하면 적용하기 어려운 문제가 발생한다. 이의 대안으로, 스펙트럼 분해를 계산하지 않고도 데이터 집합의 크기에 영향을 받지 않으면서 컴뮤트 타임을 근사적으로 계산하는 방식을 구현하고 이들의 특성을 실험적으로 분석한다.

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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    • 제37권4호
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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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    • 제32권1호
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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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    • 제31권9호
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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
    • 공업화학
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    • 제31권3호
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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.

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

  • 전계택;이태호장용근
    • KSBB Journal
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    • 제11권1호
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    • pp.30-36
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    • 1996
  • Cyclosporin A 고정화배양과 현탁배양의 결과를 바탕으로 각각의 배양의 경우에 따른 비성장속도의 포도당에 대한 Monod 속도식을 제안하고 그에 필요 한 매개변수들을 구하였다. 고정화 배양이 현탁배양 에 비해 높은 ${\mu}m$와 낮은 Km 값을 갖는 것으로 나 타났는데 이는 고정화 균체의 우수한 활성과 기질에 대한 높은 친화도에 기 인한 것으로 보인다. 고정상 발효의 경우, 구한 매개변수들을 담체내에서의 물질 전달 및 반응속도의 정도를 나타내는 효율인자 값을 계산하는데 이용하였다. 중요한 고정화 공정변수인 담체크기, 균체부하의 정도가 기질의 확산저항에 미 치는 영향을 고려하여 효율인자값을 계산한 결과, 적절한 담체의 크기는 반경 $100 ~ 500{\mu}m$로 나타났 다. 고정화세포배양시 담체내의 균체의 균일한 분포 및 활성도의 유지를 위해서, 적정한 담체입자크기를 결정한 후 균체부하량을 조절하여 고정화 공정을 운 영하는 것이 중요한 것요로 판명되었다.

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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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    • 제17권4호
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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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    • 제16권2호
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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.