WorldWideScience

Sample records for surface immobilized factors

  1. Immobilization of epidermal growth factor on titanium and stainless steel surfaces via dopamine treatment

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    Kang, Jeonghwa [Nano Medical Engineering Laboratory, RIKEN Advanced Science Institute, 2-1 Hirosawa, Wako, Saitama, 351-0198 (Japan); Department of Biological Sciences, Tokyo Metropolitan University, 1-1 Minami-Osawa, Tokyo, 192-0397 Japan (Japan); Sakuragi, Makoto; Shibata, Aya; Abe, Hiroshi; Kitajima, Takashi; Tada, Seiichi [Nano Medical Engineering Laboratory, RIKEN Advanced Science Institute, 2-1 Hirosawa, Wako, Saitama, 351-0198 (Japan); Mizutani, Masayoshi; Ohmori, Hitoshi [Material Fabrication Laboratory, RIKEN Advanced Science Institute, 2-1 Hirosawa, Wako, Saitama, 351-0198 (Japan); Ayame, Hirohito [Diagnostic Biochip Laboratory, RIKEN Center for Intellectual Property Strategies, 2-1 Hirosawa, Wako, Saitama, 351-0198 (Japan); Son, Tae Il [Bioscience and Biotechnology, Chung-Ang University, 40-1 San, Nae-Ri, Daeduck-myun, Ansung-si, Kyungki-do, 456-756 (Korea, Republic of); Aigaki, Toshiro [Department of Biological Sciences, Tokyo Metropolitan University, 1-1 Minami-Osawa, Tokyo, 192-0397 Japan (Japan); Ito, Yoshihiro, E-mail: y-ito@riken.jp [Nano Medical Engineering Laboratory, RIKEN Advanced Science Institute, 2-1 Hirosawa, Wako, Saitama, 351-0198 (Japan); Department of Biological Sciences, Tokyo Metropolitan University, 1-1 Minami-Osawa, Tokyo, 192-0397 Japan (Japan); Diagnostic Biochip Laboratory, RIKEN Center for Intellectual Property Strategies, 2-1 Hirosawa, Wako, Saitama, 351-0198 (Japan)

    2012-12-01

    Titanium and stainless steel were modified with dopamine for the immobilization of biomolecules, epidermal growth factor (EGF). First, the treatment of metal surfaces with a dopamine solution under different pH conditions was investigated. At higher pH, the dopamine solution turned brown and formed precipitates. Treatment of the metals with dopamine at pH 8.5 also resulted in the development of brown color at the surface of the metals. The hydrophobicity of the surfaces increased after treatment with dopamine, independently of pH. X-ray photoelectron spectroscopy revealed the formation of a significant amount of an organic layer on both surfaces at pH 8.5. According to ellipsometry measurements, the organic layer formed at pH 8.5 was about 1000 times as thick as that formed at pH 4.5. The amount of amino groups in the layer formed at pH 8.5 was also higher than that observed in the layer formed at pH 4.5. EGF molecules were immobilized onto the dopamine-treated surfaces via a coupling reaction using carbodiimide. A greater amount of EGF was immobilized on surfaces treated at pH 8.5 compared with pH 4.5. Significantly higher growth of rat fibroblast cells was observed on the two EGF-immobilized surfaces compared with non-immobilized surfaces in the presence of EGF. The present study demonstrated that metals can become bioactive via the surface immobilization of a growth factor and that the effect of the immobilized growth factor on metals was greater than that of soluble growth factor. - Highlights: Black-Right-Pointing-Pointer Epidermal growth factor was covalently immobilized on titan or stainless steel surfaces. Black-Right-Pointing-Pointer Amino groups were formed on the surfaces by the treatment and the growth factor was immobilized through amide bonds. Black-Right-Pointing-Pointer The immobilized epidermal growth factor accelerated cell proliferation more than soluble ones on the surfaces.

  2. Surface cell immobilization within perfluoroalkoxy microchannels

    Energy Technology Data Exchange (ETDEWEB)

    Stojkovič, Gorazd; Krivec, Matic [Faculty of Chemistry and Chemical Technology, University of Ljubljana, Aškerčeva 5, SI-1000 Ljubljana (Slovenia); Vesel, Alenka [Jožef Stefan Institute, Jamova cesta 39, 1000 Ljubljana (Slovenia); Marinšek, Marjan [Faculty of Chemistry and Chemical Technology, University of Ljubljana, Aškerčeva 5, SI-1000 Ljubljana (Slovenia); Žnidaršič-Plazl, Polona, E-mail: polona.znidarsic@fkkt.uni-lj.si [Faculty of Chemistry and Chemical Technology, University of Ljubljana, Aškerčeva 5, SI-1000 Ljubljana (Slovenia)

    2014-11-30

    Graphical abstract: - Highlights: • A very efficient approach for immobilization of cells into microreactors is presented. • It is applicable to various materials, including PFA and cyclic olefin (co)polymers. • It was used to immobilize different prokaryotic and eukaryotic microbes. • Cells were immobilized on the surface in high density and showed good stability. • Mechanisms of APTES interactions with target materials are proposed. - Abstract: Perfluoroalkoxy (PFA) is one of the most promising materials for the fabrication of cheap, solvent resistant and reusable microfluidic chips, which have been recently recognized as effective tools for biocatalytic process development. The application of biocatalysts significantly depends on efficient immobilization of enzymes or cells within the reactor enabling long-term biocatalyst use. Functionalization of PFA microchannels by 3-aminopropyltriethoxysilane (ATPES) and glutaraldehyde was used for rapid preparation of microbioreactors with surface-immobilized cells. X-ray photoelectron spectroscopy and scanning electron microscopy were used to accurately monitor individual treatment steps and to select conditions for cell immobilization. The optimized protocol for Saccharomyces cerevisiae immobilization on PFA microchannel walls comprised ethanol surface pretreatment, 4 h contacting with 10% APTES aqueous solution, 10 min treatment with 1% glutaraldehyde and 20 min contacting with cells in deionized water. The same protocol enabled also immobilization of Escherichia coli, Pseudomonas putida and Bacillus subtilis cells on PFA surface in high densities. Furthermore, the developed procedure has been proved to be very efficient also for surface immobilization of tested cells on other materials that are used for microreactor fabrication, including glass, polystyrene, poly (methyl methacrylate), polycarbonate, and two olefin-based polymers, namely Zeonor{sup ®} and Topas{sup ®}.

  3. Immobilization of Active Bacteriophages on Polyhydroxyalkanoate Surfaces.

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    Wang, Chanchan; Sauvageau, Dominic; Elias, Anastasia

    2016-01-20

    A rapid, efficient technique for the attachment of bacteriophages (phages) onto polyhydroxyalkanoate (PHA) surfaces has been developed and compared to three reported methods for phage immobilization. Polymer surfaces were modified to facilitate phage attachment using (1) plasma treatment alone, (2) plasma treatment followed by activation by 1-ethyl-3-(3-(dimethylamino)propyl)carbodiimide hydrochloride (EDC) and N-hydroxysulfosuccinimide (sulfo-NHS), (3) plasma-initiated acrylic acid grafting, or (4) plasma-initiated acrylic acid grafting with activation by EDC and sulfo-NHS. The impact of each method on the surface chemistry of PHA was investigated using contact angle analysis and X-ray photoelectron spectroscopy. Each of the four treatments was shown to result in both increased hydrophilicity and in the modification of the surface functional groups. Modified surfaces were immersed in suspensions of phage T4 for immobilization. The highest level of phage binding was observed for the surfaces modified by plasma treatment alone. The change in chemical bond states observed for surfaces that underwent plasma treatment is suspected to be the cause of the increased binding of active phages. Plasma-treated surfaces were further analyzed through phage-staining and fluorescence microscopy to assess the surface density of immobilized phages and their capacity to capture hosts. The infective capability of attached phages was confirmed by exposing the phage-immobilized surfaces to the host bacteria Escherichia coli in both plaque and infection dynamic assays. Plasma-treated surfaces with immobilized phages displayed higher infectivity than surfaces treated with other methods; in fact, the equivalent initial multiplicity of infection was 2 orders of magnitude greater than with other methods. Control samples - prepared by immersing polymer surfaces in phage suspensions (without prior plasma treatment) - did not show any bacterial growth inhibition, suggesting they did not bind

  4. Bed Rest and Immobilization: Risk Factors for Bone Loss

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    ... Risk Factors for Bone Loss Bed Rest and Immobilization: Risk Factors for Bone Loss Like muscle, bone ... complications of pregnancy; and those who are experiencing immobilization of some part of the body because of ...

  5. Drop drying on surfaces determines chemical reactivity - the specific case of immobilization of oligonucleotides on microarrays

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    2013-01-01

    Background Drop drying is a key factor in a wide range of technical applications, including spotted microarrays. The applied nL liquid volume provides specific reaction conditions for the immobilization of probe molecules to a chemically modified surface. Results We investigated the influence of nL and μL liquid drop volumes on the process of probe immobilization and compare the results obtained to the situation in liquid solution. In our data, we observe a strong relationship between drop drying effects on immobilization and surface chemistry. In this work, we present results on the immobilization of dye labeled 20mer oligonucleotides with and without an activating 5′-aminoheptyl linker onto a 2D epoxysilane and a 3D NHS activated hydrogel surface. Conclusions Our experiments identified two basic processes determining immobilization. First, the rate of drop drying that depends on the drop volume and the ambient relative humidity. Oligonucleotides in a dried spot react unspecifically with the surface and long reaction times are needed. 3D hydrogel surfaces allow for immobilization in a liquid environment under diffusive conditions. Here, oligonucleotide immobilization is much faster and a specific reaction with the reactive linker group is observed. Second, the effect of increasing probe concentration as a result of drop drying. On a 3D hydrogel, the increasing concentration of probe molecules in nL spotting volumes accelerates immobilization dramatically. In case of μL volumes, immobilization depends on whether the drop is allowed to dry completely. At non-drying conditions, very limited immobilization is observed due to the low oligonucleotide concentration used in microarray spotting solutions. The results of our study provide a general guideline for microarray assay development. They allow for the initial definition and further optimization of reaction conditions for the immobilization of oligonucleotides and other probe molecule classes to different

  6. An overview of technologies for immobilization of enzymes and surface analysis techniques for immobilized enzymes

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    Mohamad, Nur Royhaila; Marzuki, Nur Haziqah Che; Buang, Nor Aziah; Huyop, Fahrul; Wahab, Roswanira Abdul

    2015-01-01

    The current demands of sustainable green methodologies have increased the use of enzymatic technology in industrial processes. Employment of enzyme as biocatalysts offers the benefits of mild reaction conditions, biodegradability and catalytic efficiency. The harsh conditions of industrial processes, however, increase propensity of enzyme destabilization, shortening their industrial lifespan. Consequently, the technology of enzyme immobilization provides an effective means to circumvent these concerns by enhancing enzyme catalytic properties and also simplify downstream processing and improve operational stability. There are several techniques used to immobilize the enzymes onto supports which range from reversible physical adsorption and ionic linkages, to the irreversible stable covalent bonds. Such techniques produce immobilized enzymes of varying stability due to changes in the surface microenvironment and degree of multipoint attachment. Hence, it is mandatory to obtain information about the structure of the enzyme protein following interaction with the support surface as well as interactions of the enzymes with other proteins. Characterization technologies at the nanoscale level to study enzymes immobilized on surfaces are crucial to obtain valuable qualitative and quantitative information, including morphological visualization of the immobilized enzymes. These technologies are pertinent to assess efficacy of an immobilization technique and development of future enzyme immobilization strategies. PMID:26019635

  7. Immobilizing live Escherichia coli for AFM studies of surface dynamics

    International Nuclear Information System (INIS)

    Lonergan, N.E.; Britt, L.D.; Sullivan, C.J.

    2014-01-01

    Atomic force microscopy (AFM) is a probe-based technique that permits high resolution imaging of live bacterial cells. However, stably immobilizing cells to withstand the probe-based lateral forces remains an obstacle in AFM mediated studies, especially those of live, rod shaped bacteria in nutrient media. Consequently, AFM has been under-utilized in the research of bacterial surface dynamics. The aim of the current study was to immobilize a less adherent Escherichia coli strain in a method that both facilitates AFM imaging in nutrient broth and preserves overall cell viability. Immobilization reagents and buffers were systematically evaluated and the cell membrane integrity was monitored in all sample preparations. As expected, the biocompatible gelatin coated surfaces facilitated stable cell attachment in lower ionic strength buffers, yet poorly immobilized cells in higher ionic strength buffers. In comparison, poly-L-lysine surfaces bound cells in both low and high ionic strength buffers. The benefit of the poly-L-lysine binding capacity was offset by the compromised membrane integrity exhibited by cells on poly-L-lysine surfaces. However, the addition of divalent cations and glucose to the immobilization buffer was found to mitigate this unfavorable effect. Ultimately, immobilization of E. coli cells on poly-L-lysine surfaces in a lower ionic strength buffer supplemented with Mg 2+ and Ca 2+ was determined to provide optimal cell attachment without compromising the overall cell viability. Cells immobilized in this method were stably imaged in media through multiple division cycles. Furthermore, permeability assays indicated that E. coli cells recover from the hypoosmotic stress caused by immobilization in low ionic strength buffers. Taken together, this data suggests that stable immobilization of viable cells on poly-L-lysine surfaces can be accomplished in lower ionic strength buffers that are supplemented with divalent cations for membrane stabilization while

  8. Immobilization of type-I collagen and basic fibroblast growth factor (bFGF) onto poly (HEMA-co-MMA) hydrogel surface and its cytotoxicity study.

    Science.gov (United States)

    Yan, Tuo; Sun, Rong; Li, Chun; Tan, Baihua; Mao, Xuan; Ao, Ningjian

    2010-08-01

    Type-I collagen and bFGF were immobilized onto the surface of poly (HEMA-co-MMA) hydrogel by grafting and coating methods to improve its cytotoxicity. The multi-layered structure of the biocompatible layer was confirmed by FTIR, AFM and static water contact angles. The layers were stable in body-like environment (pH 7.4). Human skin fibroblast cells (HSFC) were seeded onto Col/bFGF-poly (HEMA-co-MMA), Col-poly (HEMA-co-MMA) and poly (HEMA-co-MMA) films for 1, 3 and 5 day. MTT assay was performed to evaluate the extraction toxicity of the materials. Results showed that the cell attachment, proliferation and differentiation on Col/bFGF-poly (HEMA-co-MMA) film were higher than those of the control group, which indicated the improvement of cell-material interaction. The extraction toxicity of the modified materials was also lower than that of the unmodified group. The protein and bFGF immobilized poly (HEMA-co-MMA) hydrogel might hold great promise to be a biocompatible material.

  9. Rational surface silane modification for immobilizing glucose oxidase.

    Science.gov (United States)

    Tian, Feibao; Guo, Yi; Lin, Feifei; Zhang, Yumei; Yuan, Qipeng; Liang, Hao

    2016-06-01

    Glucose oxidase (GOx) has many significant applications in biosensor and biocatalysis. In this study, we firstly quantitatively analyzed the binding efficiency of (3-aminopropyl) trimethoxysilane (APTES) modified onto the surface of GOx. It was found that the contents of the grafted silane did not significantly influence the relative activities and tertiary structures of all surface modified GOxs. Immobilization ratio and relative activity of all instances of APTES modified GOx increased, compared with those of native enzyme. However, good stability of immobilized GOx at extreme pH and high temperature could only be obtained when modified protein with low binding silane content. At pH 2.0, the immobilized GOx with low binding content showed a more than 600% activity, compared to the free enzyme. Therefore, rational surface modification would be beneficial to improving the activity and stability of immobilized enzyme as well as increasing loading amount. Copyright © 2016 Elsevier B.V. All rights reserved.

  10. A new route for chitosan immobilization onto polyethylene surface

    Czech Academy of Sciences Publication Activity Database

    Popelka, A.; Novák, I.; Lehocký, M.; Junkar, I.; Mozetič, M.; Kleinová, A.; Janigová, I.; Šlouf, Miroslav; Bílek, F.; Chodák, I.

    2012-01-01

    Roč. 90, č. 4 (2012), s. 1501-1508 ISSN 0144-8617 Institutional support: RVO:61389013 Keywords : immobilization * plasma treatment * chitosan Subject RIV: CD - Macromolecular Chemistry Impact factor: 3.479, year: 2012

  11. Optimization of Phospholipase A1 Immobilization on Plasma Surface Modified Chitosan Nanofibrous Mat

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    Zahra Beig Mohammadi

    2016-01-01

    Full Text Available Phospholipase A1 is known as an effective catalyst for hydrolysis of various phospholipids in enzymatic vegetable oil degumming. Immobilization is one of the most efficient strategies to improve its activity, recovery and functional properties. In this study, chitosan-co-polyethylene oxide (90:10 nanofibrous mat was successfully fabricated and modified with atmospheric plasma at different times (2, 6 and 10 min to interact with enzyme molecules. Scanning electron microscopy images revealed that the membranes retained uniform nanofibrous and open porous structures before and after the treatment. PLA1 was successfully immobilized onto the membrane surfaces via covalent bonds with the functional groups of chitosan nanofibrous mat. Response surface methodology was used to optimize the immobilization conditions for reaching the maximum immobilization efficiency. Enzyme concentration, pH, and immobilization time were found to be significant key factors. Under optimum conditions (5.03 h, pH 5.63, and enzyme dosage 654.36 UI, the atmospheric plasma surface modified chitosan nanofibers reached the highest immobilization efficiency (78.50%. Fourier transform infrared spectroscopy of the control and plasma surface-modified chitosan nanofibers revealed the functional groups of nanofibers and their reaction with the enzyme. The results indicated that surface modification by atmospheric plasma induced an increase in PLA1 loading on the membrane surfaces.

  12. Immobilized enzymes: understanding enzyme - surface interactions at the molecular level.

    Science.gov (United States)

    Hoarau, Marie; Badieyan, Somayesadat; Marsh, E Neil G

    2017-11-22

    Enzymes immobilized on solid supports have important and industrial and medical applications. However, their uses are limited by the significant reductions in activity and stability that often accompany the immobilization process. Here we review recent advances in our understanding of the molecular level interactions between proteins and supporting surfaces that contribute to changes in stability and activity. This understanding has been facilitated by the application of various surface-sensitive spectroscopic techniques that allow the structure and orientation of enzymes at the solid/liquid interface to be probed, often with monolayer sensitivity. An appreciation of the molecular interactions between enzyme and surface support has allowed the surface chemistry and method of enzyme attachement to be fine-tuned such that activity and stability can be greatly enhanced. These advances suggest that a much wider variety of enzymes may eventually be amenable to immobilization as green catalysts.

  13. Gelatin Functionalization of Biomaterial Surfaces: Strategies for Immobilization and Visualization

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    Peter Dubruel

    2011-01-01

    Full Text Available In the present work, the immobilization of gelatin as biopolymer on two types of implantable biomaterials, polyimide and titanium, was compared. Both materials are known for their biocompatibility while lacking cell-interactive behavior. For both materials, a pre-functionalization step was required to enable gelatin immobilization. For the polyimide foils, a reactive succinimidyl ester was introduced first on the surface, followed by covalent grafting of gelatin. For the titanium material, methacrylate groups were first introduced on the Ti surface through a silanization reaction. The applied functionalities enabled the subsequent immobilization of methacrylamide modified gelatin. Both surface modified materials were characterized in depth using atomic force microscopy, static contact angle measurements, confocal fluorescence microscopy, attenuated total reflection infrared spectroscopy and X-ray photo-electron spectroscopy. The results indicated that the strategies elaborated for both material classes are suitable to apply stable gelatin coatings. Interestingly, depending on the material class studied, not all surface analysis techniques are applicable.

  14. Oligonucleotide microarrays: immobilization of phosphorylated oligonucleotides on epoxylated surface.

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    Mahajan, S; Kumar, P; Gupta, K C

    2006-01-01

    A facile and efficient method for direct immobilization of phosphorylated oligonucleotides on an epoxy-activated glass surface is described. The new immobilization strategy has been analyzed for its performance in DNA microarray under both microwave and thermal conditions. It reflects high immobilization efficiency ( approximately 23%), and signal-to-noise ratio ( approximately 98) and resulted in high hybridization efficiency ( approximately 36%) in comparison to those obtained with standard methods, viz., NTMTA ( approximately 9.76%) and epoxide-amine ( approximately 9.82%). The probes immobilized through the new strategy were found to be heat-stable, since the performance of microarray decreased by only approximately 7% after subjecting it to 20 PCR-like heat cycles, suggesting that the chemistry could be used in integrated PCR/microarray devices. The immobilization of probes following the proposed chemistry resulted in spots of superior quality in terms of spot morphology, spot homogeneity, and signal reproducibility. The constructed microarrays have been successfully used for the discrimination of nucleotide mismatches. In conclusion, these features make the new immobilization strategy ideal for facile, efficient, and cost-effective manufacturing of DNA microarrays.

  15. Orientation control of photo-immobilized antibodies on the surface of azobenzene-containing polymers by the introduction of functional groups.

    Science.gov (United States)

    Mouri, Makoto; Ikawa, Taiji; Narita, Mamiko; Hoshino, Fumihiko; Watanabe, Osamu

    2010-06-11

    In our photo-induced immobilization technique for an antibody (IgG) using azopolymers, the introduction of COOH and NMe(2) into the azopolymers, which can introduce surface charges, strongly affected the immobilization properties such as the efficiency of immobilization and the activity of the immobilized IgG (i.e., the orientation of the immobilized IgG). The introduction of COOH promoted a more active orientation of the immobilized IgG. The orientation was determined during the adsorption process onto the azopolymer surface in solution before photo-immobilization, and was maintained during the photo-immobilization. The surface charge of the azopolymer appears to be an important factor for IgG orientation, which involves electrostatic interactions between its Fab and the azopolymer surface.

  16. Covalent immobilization of antimicrobial peptides (AMPs) onto biomaterial surfaces.

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    Costa, Fabíola; Carvalho, Isabel F; Montelaro, Ronald C; Gomes, P; Martins, M Cristina L

    2011-04-01

    Bacterial adhesion to biomaterials remains a major problem in the medical devices field. Antimicrobial peptides (AMPs) are well-known components of the innate immune system that can be applied to overcome biofilm-associated infections. Their relevance has been increasing as a practical alternative to conventional antibiotics, which are declining in effectiveness. The recent interest focused on these peptides can be explained by a group of special features, including a wide spectrum of activity, high efficacy at very low concentrations, target specificity, anti-endotoxin activity, synergistic action with classical antibiotics, and low propensity for developing resistance. Therefore, the development of an antimicrobial coating with such properties would be worthwhile. The immobilization of AMPs onto a biomaterial surface has further advantages as it also helps to circumvent AMPs' potential limitations, such as short half-life and cytotoxicity associated with higher concentrations of soluble peptides. The studies discussed in the current review report on the impact of covalent immobilization of AMPs onto surfaces through different chemical coupling strategies, length of spacers, and peptide orientation and concentration. The overall results suggest that immobilized AMPs may be effective in the prevention of biofilm formation by reduction of microorganism survival post-contact with the coated biomaterial. Minimal cytotoxicity and long-term stability profiles were obtained by optimizing immobilization parameters, indicating a promising potential for the use of immobilized AMPs in clinical applications. On the other hand, the effects of tethering on mechanisms of action of AMPs have not yet been fully elucidated. Therefore, further studies are recommended to explore the real potential of immobilized AMPs in health applications as antimicrobial coatings of medical devices. Copyright © 2010 Acta Materialia Inc. All rights reserved.

  17. Surface-immobilized DNAzyme-type biocatalysis

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    Stefan, Loic; Lavergne, Thomas; Spinelli, Nicolas; Defrancq, Eric; Monchaud, David

    2014-02-01

    The structure of the double helix of deoxyribonucleic acid (DNA, also called duplex-DNA) was elucidated sixty years ago by Watson, Crick, Wilkins and Franklin. Since then, DNA has continued to hold a fascination for researchers in diverse fields including medicine and nanobiotechnology. Nature has indeed excelled in diversifying the use of DNA: beyond its canonical role of repository of genetic information, DNA could also act as a nanofactory able to perform some complex catalytic tasks in an enzyme-mimicking manner. The catalytic capability of DNA was termed DNAzyme; in this context, a peculiar DNA structure, a quadruple helix also named quadruplex-DNA, has recently garnered considerable interest since its autonomous catalytic proficiency relies on its higher-order folding that makes it suitable to interact efficiently with hemin, a natural cofactor of many enzymes. Quadruplexes have thus been widely studied for their hemoprotein-like properties, chiefly peroxidase-like activity, i.e., their ability to perform hemin-mediated catalytic oxidation reactions. Recent literature is replete with applications of quadruplex-based peroxidase-mimicking DNAzyme systems. Herein, we take a further leap along the road to biochemical applications, assessing the actual efficiency of catalytic quadruplexes for the detection of picomolar levels of surface-bound analytes in an enzyme-linked immunosorbent (ELISA)-type assay. To this end, we exploit an innovative strategy based on the functionalization of DNA by a multitasking platform named RAFT (for regioselectivity addressable functionalized template), whose versatility enables the grafting of DNA whatever its nature (duplex-DNA, quadruplex-DNA, etc.). We demonstrate that the resulting biotinylated RAFT/quadruplex systems indeed acquire catalytic properties that allow for efficient luminescent detection of picomoles of surface-bound streptavidin. We also highlight some of the pitfalls that have to be faced during optimization

  18. Self-assembling peptide hydrogels immobilized on silicon surfaces

    International Nuclear Information System (INIS)

    Franchi, Stefano; Battocchio, Chiara; Galluzzi, Martina; Navisse, Emanuele; Zamuner, Annj; Dettin, Monica; Iucci, Giovanna

    2016-01-01

    The hydrogels of self-assembling ionic complementary peptides have collected in the scientific community increasing consensus as mimetics of the extracellular matrix that can offer 3D supports for cell growth or be vehicles for the delivery of stem cells or drugs. Such scaffolds have also been proposed as bone substitutes for small defects as they promote beneficial effects on human osteoblasts. In this context, our research deals with the introduction of a layer of self-assembling peptides on a silicon surface by covalent anchoring and subsequent physisorption. In this work, we present a spectroscopic investigation of the proposed bioactive scaffolds, carried out by surface-sensitive spectroscopic techniques such as XPS (X-ray photoelectron spectroscopy) and RAIRS (Reflection Absorption Infrared Spectroscopy) and by state-of-the-art synchrotron radiation methodologies such as angle dependent NEXAFS (Near Edge X-ray Absorption Fine Structure). XPS studies confirmed the change in the surface composition in agreement with the proposed enrichments, and led to assess the self-assembling peptide chemical stability. NEXAFS spectra, collected in angular dependent mode at the N K-edge, allowed to investigate the self-assembling behavior of the macromolecules, as well as to determine their molecular orientation on the substrate. Furthermore, Infrared Spectroscopy measurements demonstrated that the peptide maintains its secondary structure (β-sheet anti-parallel) after deposition on the silicon surface. The complementary information acquired by means of XPS, NEXAFS and RAIRS lead to hypothesize a “layer-by-layer” arrangement of the immobilized peptides, giving rise to an ordered 3D nanostructure. - Highlights: • A self-assembling peptide (SAP) was covalently immobilized of on a flat silicon surface. • A physisorbed SAP layer was grown on top of the covalently immobilized peptide layer. • Molecular order and orientation of the peptide overlayer on the flat silicon

  19. High Efficiency Acetylcholinesterase Immobilization on DNA Aptamer Modified Surfaces

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    Orada Chumphukam

    2014-04-01

    Full Text Available We report here the in vitro selection of DNA aptamers for electric eel acetylcholinesterase (AChE. One selected aptamer sequence (R15/19 has a high affinity towards the enzyme (Kd = 157 ± 42 pM. Characterization of the aptamer showed its binding is not affected by low ionic strength (~20 mM, however significant reduction in affinity occurred at high ionic strength (~1.2 M. In addition, this aptamer does not inhibit the catalytic activity of AChE that we exploit through immobilization of the DNA on a streptavidin-coated surface. Subsequent immobilization of AChE by the aptamer results in a 4-fold higher catalytic activity when compared to adsorption directly on to plastic.

  20. Analytical performance of molecular beacons on surface immobilized gold nanoparticles of varying size and density.

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    Uddayasankar, Uvaraj; Krull, Ulrich J

    2013-11-25

    The high quenching efficiency of metal nanoparticles has facilitated its use as quenchers in molecular beacons. To optimize this system, a good understanding of the many factors that influence molecular beacon performance is required. In this study, molecular beacon performance was evaluated as a function of gold nanoparticle size and its immobilization characteristics. Gold nanoparticles of 4 nm, 15 nm and 87 nm diameter, were immobilized onto glass slides. Each size regime offered distinctive optical properties for fluorescence quenching of molecular dyes that were conjugated to oligonucleotides that were immobilized to the gold nanoparticles. Rigid double stranded DNA was used as a model to place fluorophores at different distances from the gold nanoparticles. The effect of particle size and also the immobilization density of nanoparticles was evaluated. The 4 nm and 87 nm gold nanoparticles offered the highest sensitivity in terms of the change in fluorescence intensity as a function of distance (3-fold improvement for Cy5). The optical properties of the molecular fluorophore was of significance, with Cy5 offering higher contrast ratios than Cy3 due to the red-shifted emission spectrum relative to the plasmon peak. A high density of gold nanoparticles reduced contrast ratios, indicating preference for a monolayer of immobilized nanoparticles when considering analytical performance. Molecular beacon probes were then used in place of the double stranded oligonucleotides. There was a strong dependence of molecular beacon performance on the length of a linker used for attachment to the nanoparticle surface. The optimal optical performance was obtained with 4 nm gold nanoparticles that were immobilized as monolayers of low density (5.7×10(11)particles cm(-2)) on glass surfaces. These nanoparticle surfaces offered a 2-fold improvement in analytical performance of the molecular beacons when compared to other nanoparticle sizes investigated. The principles developed

  1. Surface coordination polymerization of ethylene by hydrozirconation-immobilized metallocene.

    Science.gov (United States)

    Zheng, Jun; Wang, Yanhui; Ye, Lin; Lin, Yichao; Tang, Tao; Zhang, Jidong

    2014-07-01

    Hydrozirconation on vinyl-terminated substrates (silicon wafer and nanosilica sphere) is employed as an efficient way for immobilization of zirconocene catalyst through Zr-C bonds, which is applied in surface coordination ethylene polymerization producing surface-tethered polyethylene (PE). The formation of Zr-C σ bond induced by hydrozirconation provides an initiator precursor for growing a layer of PE covalently linked onto substrates. The results from SEM, AFM, and TEM show that the surface polymerization is controlled by hydrozirconation. Surface pattern or core-shell structure with crystalline PE coating can be formed, when silicon wafer is selectively functionalized with vinyl-groups or vinyl-modified nanosilica is applied. It is believed that hydrozirconation for the synthesis of zirconocene initiator can be a versatile route to prepare polyolefin hybrid materials. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  2. Influence of cosolvents on the hydrophobic surface immobilization topography of Candida antarctica lipase B

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    The presence of cosolvents and co-solutes during the immobilization of lipases on hydrophobic supports may influence the extent of lipase immobilization and the long-term catalytic stability of the biocatalyst. Candida antarctica B lipase immobilization was examined on a hydrophobic surface, i.e., ...

  3. Chitosan Immobilized on Silica Surface for Wastewater Treatment

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    Tetyana BUDNYAK

    2014-06-01

    Full Text Available Adsorbents of natural origin are widely used for industrial wastewater treatment in order to replace current costly methods of heavy metals removing from solutions. For that purpose, immobilization of chitosan on the silica gel surface by crosslinking of adsorbed biopolymer with glutaraldehyde was carried out. Anchorage of chitosan on the silica surface was confirmed by IR spectroscopy. Concentration of immobilized chitosan was estimated using thermogravimetric analysis. The adsorption of microquantities of Zn(II, Cu(II, Cd(II, Pb(II, Fe(III, V(V and Mo(VI ions from aqueous solutions by obtained composite has been studied. The highest sorption capacities were observed with respect to zinc (0.46 mmol/g, molybdenum- and vanadium-containing ions (0.31 mmol/g. Diffuse reflectance electronic spectra of samples with different content of adsorbed metal and kinetic adsorption characteristics of the composite adsorbent were estimated. DOI: http://dx.doi.org/10.5755/j01.ms.20.2.4975

  4. Electrochemical immobilization of biomolecules on gold surface modified with monolayered L-cysteine

    International Nuclear Information System (INIS)

    Honda, Mitsunori; Baba, Yuji; Sekiguchi, Tetsuhiro; Shimoyama, Iwao; Hirao, Norie

    2014-01-01

    Immobilization of organic molecules on the top of a metal surface is not easy because of lattice mismatch between organic and metal crystals. Gold atoms bind to thiol groups through strong chemical bonds, and a self-assembled monolayer of sulfur-terminated organic molecules is formed on the gold surface. Herein, we suggested that a monolayer of L-cysteine deposited on a gold surface can act as a buffer layer to immobilize biomolecules on the metal surface. We selected lactic acid as the immobilized biomolecule because it is one of the simplest carboxyl-containing biomolecules. The immobilization of lactic acid on the metal surface was carried out by an electrochemical method in an aqueous environment under the potential range varying from − 0.6 to + 0.8 V. The surface chemical states before and after the electrochemical reaction were characterized using X-ray photoelectron spectroscopy (XPS). The N 1s and C 1s XPS spectra showed that the L-cysteine-modified gold surface can immobilize lactic acid via peptide bonds. This technique might enable the immobilization of large organic molecules and biomolecules. - Highlights: • Monolayer l-cysteine deposited on Au surface as a buffer layer to immobilize biomolecules. • Lactic acid as the immobilized biomolecule as it is simple carboxyl-containing biomolecule. • X-ray photoelectron spectroscopy (XPS) of surface chemical states, before and after. • L-cysteine-modified Au surface can immobilize lactic acid via peptide bonds

  5. Optimizations of α-amylase production by response surface methodology in immobilization Bacillus amyloliquefaciens ATCC 23350

    Directory of Open Access Journals (Sweden)

    Hamid reza Samadlouie

    2016-03-01

    Full Text Available Introduction: Production of an endogenous α-amylase from Bacillus amyloliquefaciens ATCC 23350 was studied and enhanced. Materials and methods: Protein and carbon sources were analyzed for free and immobilized bacterial cells and number of beads was considered for immobilized cells via one factor at a time methodforα-amylase production by Bacillus amyloliquefaciens. Subsequently, optimization condition was employed solely for immobilized bacterial cells by response surface methodology (RSM. Results: Peptone and rice starch showed to improve the α-amylase production in immobilized Bacillus cells. RSM generated a mathematical model explaining the optimum concentration of the efficient nutrients (139.35 g/l of rice starch and 80.00 g/l of peptone leading to an optimum amylase production (205 U/ml. Discussion and conclusion: The statistical advance displayed significant outcomes to optimize the process parameters for maximal α-amylase production using Bacillus amyloliquefaciens and gave permission to rapid screening of variables. RSM led to find out an immense improvement in enzyme activity (more than 90%: from 25 to 225 U/ml for the first time. 

  6. Thermolysis of surface-immobilized phenethyl phenyl ether

    Energy Technology Data Exchange (ETDEWEB)

    Britt, P.F.; Buchanan, A.C. III; Hitsman, V.M.

    1991-01-01

    Our research has focused on modeling the constraints on free-radical reactions that might be imposed in coal as a consequence of its cross-linked macromolecular structure by covalently bonding diphenylalkanes to an inert silica surface. A surface-immobilized phenethyl phenyl ether ({approx}PhCH{sub 2}CH{sub 2}POh, or {approx}PPE-3) has been prepared as a model for ether linkages in lignin by the condensation of p-HOPhCH{sub 2}CH{sub 2}OPh with the surface hydroxyls of a high purity fumed silica. Thermolysis of {approx}PPE-3 at saturation surface coverage at 375{degree}C produces {approx}PhCH = CH{sub 2} and PhOH as the major products which are consistent with the proposed free-radical chain mechanism for the decomposition of fluid-phase phenethyl phenyl ether. However, significant quantities of {approx}PhCH{sub 3} and PhCHO (ca. 18% of the products) are produced indicating the emergence of a new reaction pathway on the surface. The mechanism for the decomposition of {approx}PPE-3 will be discussed in light of this new information. 18 refs., 1 fig.

  7. Immobilization of biomolecules onto surfaces according to ultraviolet light diffraction patterns

    DEFF Research Database (Denmark)

    Petersen, Steffen B.; Gennaro, Ane Kold Di; Neves Petersen, Teresa

    2010-01-01

    We developed a method for immobilization of biomolecules onto thiol functionalized surfaces according to UV diffraction patterns. UV light-assisted molecular immobilization proceeds through the formation of free, reactive thiol groups that can bind covalently to thiol reactive surfaces. We...... demonstrate that, by shaping the pattern of the UV light used to induce molecular immobilization, one can control the pattern of immobilized molecules onto the surface. Using a single-aperture spatial mask, combined with the Fourier transforming property of a focusing lens, we show that submicrometer (0.7 mu...

  8. Zwitterionic sulfobetaine polymer-immobilized surface by simple tyrosinase-mediated grafting for enhanced antifouling property.

    Science.gov (United States)

    Kwon, Ho Joon; Lee, Yunki; Phuong, Le Thi; Seon, Gyeung Mi; Kim, Eunsuk; Park, Jong Chul; Yoon, Hyunjin; Park, Ki Dong

    2017-10-01

    Introducing antifouling property to biomaterial surfaces has been considered an effective method for preventing the failure of implanted devices. In order to achieve this, the immobilization of zwitterions on biomaterial surfaces has been proven to be an excellent way of improving anti-adhesive potency. In this study, poly(sulfobetaine-co-tyramine), a tyramine-conjugated sulfobetaine polymer, was synthesized and simply grafted onto the surface of polyurethane via a tyrosinase-mediated reaction. Surface characterization by water contact angle measurements, X-ray photoelectron spectroscopy and atomic force microscopy demonstrated that the zwitterionic polymer was successfully introduced onto the surface of polyurethane and remained stable for 7days. In vitro studies revealed that poly(sulfobetaine-co-tyramine)-coated surfaces dramatically reduced the adhesion of fibrinogen, platelets, fibroblasts, and S. aureus by over 90% in comparison with bare surfaces. These results proved that polyurethane surfaces grafted with poly(sulfobetaine-co-tyramine) via a tyrosinase-catalyzed reaction could be promising candidates for an implantable medical device with excellent bioinert abilities. Antifouling surface modification is one of the key strategy to prevent the thrombus formation or infection which occurs on the surface of biomaterial after transplantation. Although there are many methods to modify the surface have been reported, necessity of simple modification technique still exists to apply for practical applications. The purpose of this study is to modify the biomaterial's surface by simply immobilizing antifouling zwitterion polymer via enzyme tyrosinase-mediated reaction which could modify versatile substrates in mild aqueous condition within fast time period. After modification, pSBTA grafted surface becomes resistant to various biological factors including proteins, cells, and bacterias. This approach appears to be a promising method to impart antifouling property on

  9. Influence of acetylcholinesterase immobilization on the photoluminescence properties of mesoporous silicon surface

    International Nuclear Information System (INIS)

    Saleem, Muhammad; Rafiq, Muhammad; Seo, Sung-Yum; Lee, Ki Hwan

    2014-01-01

    Acetylcholinesterase immobilized p-type porous silicon surface was prepared by covalent attachment. The immobilization procedure was based on support surface chemical oxidation, silanization, surface activation with cyanuric chloride and finally covalent attachment of free enzyme on the cyanuric chloride activated porous silicon surface. Different pore diameter of porous silicon samples were prepared by electrochemical etching in HF based electrolyte solution and appropriate sample was selected suitable for enzyme immobilization with maximum trapping ability. The surface modification was studied through field emission scanning electron microscope, EDS, FT-IR analysis, and photoluminescence measurement by utilizing the fluctuation in the photoluminescence of virgin and enzyme immobilized porous silicon surface. Porous silicon showed strong photoluminescence with maximum emission at 643 nm and immobilization of acetylcholinesterase on porous silicon surface cause considerable increment on the photoluminescence of porous silicon material while acetylcholinesterase free counterpart did not exhibit any fluorescence in the range of 635–670 nm. The activities of the free and immobilized enzymes were evaluated by spectrophotometric method by using neostigmine methylsulfate as standard enzyme inhibitor. The immobilized enzyme exhibited considerable response toward neostigmine methylsulfate in a dose dependent manner comparable with that of its free counterpart alongside enhanced stability, easy separation from the reaction media and significant saving of enzyme. It was believed that immobilized enzyme can be exploited in organic and biomolecule synthesis possessing technical and economical prestige over free enzyme and prominence of easy separation from the reaction mixture.

  10. Immobilization of biomolecules onto surfaces according to ultraviolet light diffraction patterns

    International Nuclear Information System (INIS)

    Bjoern Petersen, Steffen; Kold di Gennaro, Ane; Neves-Petersen, Maria Teresa; Skovsen, Esben; Parracino, Antonietta

    2010-01-01

    We developed a method for immobilization of biomolecules onto thiol functionalized surfaces according to UV diffraction patterns. UV light-assisted molecular immobilization proceeds through the formation of free, reactive thiol groups that can bind covalently to thiol reactive surfaces. We demonstrate that, by shaping the pattern of the UV light used to induce molecular immobilization, one can control the pattern of immobilized molecules onto the surface. Using a single-aperture spatial mask, combined with the Fourier transforming property of a focusing lens, we show that submicrometer (0.7 μm) resolved patterns of immobilized prostate-specific antigen biomolecules can be created. If a dual-aperture spatial mask is used, the results differ from the expected Fourier transform pattern of the mask. It appears as a superposition of two diffraction patterns produced by the two apertures, with a fine structured interference pattern superimposed.

  11. Active-site titration analysis of surface influence on immobilized Candida antarctica Lipase B activity

    Science.gov (United States)

    Matrix morphology and surface polarity effects were investigated for Candida antarctica lipase B immobilization. Measurements of the amount of lipase immobilized (bicinchoninic acid method) and the catalyst’s tributyrin hydrolysis activity, coupled with a determination of the lipase’s functional fr...

  12. Chondroitin sulfate immobilization at the surface of electrospun nanofiber meshes for cartilage tissue regeneration approaches

    Energy Technology Data Exchange (ETDEWEB)

    Piai, Juliana Francis [3B’s Research Group − Biomaterials, Biodegradables and Biomimetics, Department of Polymer Engineering, University of Minho, Headquarters of the European Institute of Excellence on Tissue Engineering and Regenerative Medicine, AvePark, 4805-017 Barco, Guimarães (Portugal); ICVS/3B’s − PT Government Associate Laboratory, Braga/Guimarães (Portugal); Grupo de Materiais Poliméricos e Compósitos, GMPC – Departamento de Química- Universidade Estadual de Maringá, Av. Colombo 5790, 87020-900, Maringá, Paraná (Brazil); Alves da Silva, Marta; Martins, Albino; Torres, Ana Bela [3B’s Research Group − Biomaterials, Biodegradables and Biomimetics, Department of Polymer Engineering, University of Minho, Headquarters of the European Institute of Excellence on Tissue Engineering and Regenerative Medicine, AvePark, 4805-017 Barco, Guimarães (Portugal); ICVS/3B’s − PT Government Associate Laboratory, Braga/Guimarães (Portugal); Faria, Susana [Research Center Officinal Mathematical, Department of Mathematics for Science and Technology, University of Minho, Campus de Azurém, 4800-058 Guimarães (Portugal); and others

    2017-05-01

    Highlights: • Chemical immobilization of chondroitin sulfate at the surface of nanofiber meshes. • CS-immobilized NFMs showed lower roughness and higher hydrophilicity. • CS-immobilized NFMs offer a highly effective substrate for hACs phenotypic stability. - Abstract: Aiming at improving the biocompatibility of biomaterial scaffolds, surface modification presents a way to preserve their mechanical properties and to improve the surface bioactivity. In this work, chondroitin sulfate (CS) was immobilized at the surface of electrospun poly(caprolactone) nanofiber meshes (PCL NFMs), previously functionalized by UV/O{sub 3} exposure and aminolysis. Contact angle, SEM, optical profilometry, FTIR, X-ray photoelectron spectroscopy techniques confirmed the success of CS-immobilization in PCL NFMs. Furthermore, CS-immobilized PCL NFMs showed lower roughness and higher hydrophilicity than the samples without CS. Human articular chondrocytes (hACs) were cultured on electrospun PCL NFMs with or without CS immobilization. It was observed that hACs proliferated through the entire time course of the experiment in both types of nanofibrous scaffolds, as well as for the production of glycosaminoglycans. Quantitative-PCR results demonstrated over-expression of cartilage-related genes such as Aggrecan, Collagen type II, COMP and Sox9 on both types of nanofibrous scaffolds. Morphological observations from SEM and LSCM revealed that hACs maintained their characteristic round shape and cellular agglomeration exclusively on PCL NFMs with CS immobilization. In conclusion, CS immobilization at the surface of PCL NFMs was achieved successfully and provides a valid platform enabling further surface functionalization methods in scaffolds to be developed for cartilage tissue engineering.

  13. Scanning electron microscopy study of protein immobilized on SIO2 Sol-gel surfaces

    Directory of Open Access Journals (Sweden)

    Assis O.B.G.

    2003-01-01

    Full Text Available Uniform attachment of enzymes to solid surfaces is essential in the development of bio and optical sensor devices. Immobilization by adsorption according to hydrophilic or hydrophobic nature is dependent on the charges and defects of the support surfaces. Sol-gel SiO2 densified glass surfaces, frequently used as supports for protein immobilization, are evaluated via scanning electron microscopy. The model protein is globular enzyme lysozyme, deposited by adsorption on functionalized surfaces. Formation of a protein layer is confirmed by FTIR spectroscopy, and the SEM images suggest discontinuous adsorption in areas where cracks predominate on the glass surface.

  14. Magnetic catechol-chitosan with bioinspired adhesive surface: preparation and immobilization of ω-transaminase.

    Directory of Open Access Journals (Sweden)

    Kefeng Ni

    Full Text Available The magnetic chitosan nanocomposites have been studied intensively and been used practically in various biomedical and biological applications including enzyme immobilization. However, the loading capacity and the remained activity of immobilized enzyme based on existing approaches are not satisfied. Simpler and more effective immobilization strategies are needed. Here we report a simple catechol modified protocol for preparing a novel catechol-chitosan (CCS-iron oxide nanoparticles (IONPs composites carrying adhesive moieties with strong surface affinity. The ω-transaminase (ω-TA was immobilized onto this magnetic composite via nucleophilic reactions between catechol and ω-TA. Under optimal conditions, 87.5% of the available ω-TA was immobilized on the composite, yielding an enzyme loading capacity as high as 681.7 mg/g. Furthermore, the valuation of enzyme activity showed that ω-TA immobilized on CCS-IONPs displayed enhanced pH and thermal stability compared to free enzyme. Importantly, the immobilized ω-TA retained more than 50% of its initial activity after 15 repeated reaction cycles using magnetic separation and 61.5% of its initial activity after storage at 4°C in phosphate buffered saline (PBS for 15 days. The results suggested that such adhesive magnetic composites may provide an improved platform technology for bio-macromolecules immobilized.

  15. Chondroitin sulfate immobilization at the surface of electrospun nanofiber meshes for cartilage tissue regeneration approaches

    Science.gov (United States)

    Piai, Juliana Francis; da Silva, Marta Alves; Martins, Albino; Torres, Ana Bela; Faria, Susana; Reis, Rui L.; Muniz, Edvani Curti; Neves, Nuno M.

    2017-05-01

    Aiming at improving the biocompatibility of biomaterial scaffolds, surface modification presents a way to preserve their mechanical properties and to improve the surface bioactivity. In this work, chondroitin sulfate (CS) was immobilized at the surface of electrospun poly(caprolactone) nanofiber meshes (PCL NFMs), previously functionalized by UV/O3 exposure and aminolysis. Contact angle, SEM, optical profilometry, FTIR, X-ray photoelectron spectroscopy techniques confirmed the success of CS-immobilization in PCL NFMs. Furthermore, CS-immobilized PCL NFMs showed lower roughness and higher hydrophilicity than the samples without CS. Human articular chondrocytes (hACs) were cultured on electrospun PCL NFMs with or without CS immobilization. It was observed that hACs proliferated through the entire time course of the experiment in both types of nanofibrous scaffolds, as well as for the production of glycosaminoglycans. Quantitative-PCR results demonstrated over-expression of cartilage-related genes such as Aggrecan, Collagen type II, COMP and Sox9 on both types of nanofibrous scaffolds. Morphological observations from SEM and LSCM revealed that hACs maintained their characteristic round shape and cellular agglomeration exclusively on PCL NFMs with CS immobilization. In conclusion, CS immobilization at the surface of PCL NFMs was achieved successfully and provides a valid platform enabling further surface functionalization methods in scaffolds to be developed for cartilage tissue engineering.

  16. Immobilization of microbial cells on cellulose-polymer surfaces by radiation polymerization

    International Nuclear Information System (INIS)

    Kumakura, M.; Kaetsu, I.

    1983-01-01

    Streptomyces phaeochromogens cells were immobilized on cellulose-polymer surfaces by radiation polymerization using hydrophilic monomers and paper. The enzyme activity of immobilized cell sheets was higher than that of immobilized cell composites obtained by the usual radiation polymerization technique. The enzyme activity of the sheets was affected by monomer concentration, the thickness of paper, and the degree of polymerization of paper. The copolymerization of hydroxyethyl methacrylate and methoxytetraethyleneglycol methacrylate in the sheets led to a further increase of the enzyme activity due to the increase of the hydrophilicity of the polymer matrix. The Michaelis constant of the sheets from low monomer concentration was close to that of intact cells

  17. Surface modification of magnetite nanoparticles using gluconic acid and their application in immobilized lipase.

    Science.gov (United States)

    Sui, Ying; Cui, Yu; Nie, Yong; Xia, Guang-Ming; Sun, Guo-Xin; Han, Jing-Tian

    2012-05-01

    Superparamagnetic magnetite nanoparticles (SMN) were surface-modified with gluconic acid (GLA) to improve their hydrophilicity and bio-affinity. Gluconic acid was successfully coated on the surface of magnetite nanoparticles and characterized using Fourier transform infrared spectroscopy (FT-IR). With water-soluble carbodiimide (EDC) as the coupling reagent, lipase was successfully immobilized onto the hydroxyl-functionalized magnetic nanoparticles. The immobilized lipase had better resistance to temperature and pH inactivation in comparison to the free form and hence widened the reaction pH and temperature range. Thermostability and storage stability of the enzyme improved upon covalent immobilization. Immobilized lipase showed higher activity after recycling when compared to the free one and could be recovered by magnetic separation. Copyright © 2011 Elsevier B.V. All rights reserved.

  18. AFM imaging of bacteria in liquid media immobilized on gelatin coated mica surfaces

    International Nuclear Information System (INIS)

    Doktycz, M.J.; Sullivan, C.J.; Hoyt, P.R.; Pelletier, D.A.; Wu, S.; Allison, D.P.

    2003-01-01

    Immobilization of particulates, especially biomolecules and cells, onto surfaces is critical for imaging with the atomic force microscope (AFM). In this paper, gelatin coated mica surfaces are shown to be suitable for immobilizing and imaging both gram positive, Staphylococcus aureus, and gram negative, Escherichia coli, bacteria in both air and liquid environments. Gelatin coated surfaces are shown to be superior to poly-L-lysine coated surfaces that are commonly used for the immobilization of cells. This cell immobilization technique is being developed primarily for live cell imaging of Rhodopseudomonas palustris. The genome of R. palustris has been sequenced and the organism is the target of intensive studies aimed at understanding genome function. Images of R. palustris grown both aerobically and anaerobically in liquid media are presented. Images in liquid media show the bacteria is rod shaped and smooth while images in air show marked irregularity and folding of the surface. Significant differences in the vertical dimension are also apparent with the height of the bacteria in liquid being substantially greater than images taken in air. In air immobilized bacterial flagella are clearly seen while in liquid this structure is not visible. Additionally, significant morphological differences are observed that depend on the method of bacterial growth

  19. Method for selective immobilization of macromolecules on self assembled monolayer surfaces

    Science.gov (United States)

    Laskin, Julia [Richland, WA; Wang, Peng [Billerica, MA

    2011-11-29

    Disclosed is a method for selective chemical binding and immobilization of macromolecules on solid supports in conjunction with self-assembled monolayer (SAM) surfaces. Immobilization involves selective binding of peptides and other macromolecules to SAM surfaces using reactive landing (RL) of mass-selected, gas phase ions. SAM surfaces provide a simple and convenient platform for tailoring chemical properties of a variety of substrates. The invention finds applications in biochemistry ranging from characterization of molecular recognition events at the amino acid level and identification of biologically active motifs in proteins, to development of novel biosensors and substrates for stimulated protein and cell adhesion.

  20. Photochemical immobilization of anthraquinone conjugated oligonucleotides and PCR amplicons on solid surfaces

    DEFF Research Database (Denmark)

    Koch, T.; Jacobsen, N.; Fensholdt, J.

    2000-01-01

    Ligand immobilization on solid surfaces is an essential step in fields such as diagnostics, bio sensor manufacturing, and new material sciences in general. In this paper a photochemical approach based on anthraquinone as the chromophore is presented. Photochemical procedures offer special...... advantages as they are able to generate highly reactive species in an orientation specific manner. As presented here, anthraquinone (AQ) mediated covalent DNA immobilization appears to be superior to currently known procedures. A synthetic procedure providing AQ-phosphoramidites is presented. These reagents...... facilitate AQ conjugation during routine DNA synthesis, thus enabling the AQ-oligonucleotides to be immobilized in a very convenient and efficient manner. AQ-conjugated PCR primers can be used directly in PCR. When the PCR is performed in solution, the amplicons can be immobilized after the PCR. Moreover...

  1. Platelet adhesion and cellular interaction with poly(ethylene oxide) immobilized onto silicone rubber membrane surfaces.

    Science.gov (United States)

    Hsiue, G H; Lee, S D; Chang, P C

    1996-01-01

    Cellular interaction and platelet adsorption were investigated on poly(ethylene oxide) (PEO) immobilized silicone rubber membrane (SR) which has polyacrylic acid grafts on the surfaces. Polyacrylic acid (PAA) had been introduced to the SR surface after Ar plasma treatment of SR surfaces to introduce peroxide groups. Surface characterizations were made using ATR-FTIR, ESCA, SEM, and contact angle measurements. Experimental results obtained by ESCA high resolution curve fitting spectra indicated that the amount of bisamino PEO of different molecular weights immobilized onto SR surfaces were similar, which showed that the influence of the length of molecular chains (-C-C-O-) on the reactivity of terminal amino group is negligible. The wettability of modified SR surfaces increased with an increase in PEO molecular weight. Biological studies such as corneal epithelial cell culture and blood platelet adhesion were performed to understand the biocompatibility of modified SR surfaces. Biological studies using corneal epithelial cells showed that cell migration, attachment and proliferation onto PEO-20000 immobilized SR surface were suppressed, whereas these biological activities on PEO-600 were enhanced. Another study on platelet adhesion revealed that many platelets attached to PEO-600 immobilized SR, while platelet deposition was rarely observed on SR grafted with PEO-3350. The effects of different PEO molecular chains on biological response were discussed.

  2. Alteration of cartilage surface collagen fibers differs locally after immobilization of knee joints in rats

    Science.gov (United States)

    Nagai, Momoko; Aoyama, Tomoki; Ito, Akira; Tajino, Junichi; Iijima, Hirotaka; Yamaguchi, Shoki; Zhang, Xiangkai; Kuroki, Hiroshi

    2015-01-01

    The purpose of this study was to examine the ultrastructural changes of surface cartilage collagen fibers, which differ by region and the length of the experimental period in an immobilization model of rat. Male Wistar rats were randomly divided into histological or macroscopic and ultrastructural assessment groups. The left knees of all the animals were surgically immobilized by external fixation for 1, 2, 4, 8 or 16 weeks (n = 5/time point). Sagittal histological sections of the medial mid-condylar region of the knee were obtained and assessed in four specific regions (contact and peripheral regions of the femur and tibia) and two zones (superficial and deep). To semi-quantify the staining intensity of the collagen fibers in the cartilage, picrosirius red staining was used. The cartilage surface changes of all the assessed regions were investigated by scanning electron microscopy (SEM). From histological and SEM observations, the fibrillation and irregular changes of the cartilage surface were more severe in the peripheral region than in the contact region. Interestingly, at 16 weeks post-immobilization, we observed non-fibrous structures at both the contact and peripheral regions. The collagen fiber staining intensity decreased in the contact region compared with the peripheral region. In conclusion, the alteration of surface collagen fiber ultrastructure and collagen staining intensity differed by the specific cartilage regions after immobilization. These results demonstrate that the progressive degeneration of cartilage is region specific, and depends on the length of the immobilization period. PMID:25939458

  3. Concentration- and time-dependent response of human gingival fibroblasts to fibroblast growth factor 2 immobilized on titanium dental implants.

    Science.gov (United States)

    Ma, Qianli; Wang, Wei; Chu, Paul K; Mei, Shenglin; Ji, Kun; Jin, Lei; Zhang, Yumei

    2012-01-01

    Titanium (Ti) implants are widely used clinically, but peri-implantitis remains one of the most common and serious complications. Healthy integration between gingival tissue and the implant surface is critical to long-term success in dental implant therapy. The objective of this study was to investigate how different concentrations of immobilized fibroblast growth factor 2 (FGF2) on the titania nanotubular surface influence the response of human gingival fibroblasts (HGFs). Pure Ti metal was anodized at 20 V to form a vertically organized titanium dioxide nanotube array on which three concentrations of FGF2 (250 ng/mL, 500 ng/mL, or 1000 ng/mL) were immobilized by repeated lyophilization. Surface topography was observed and FGF2 elution was detected using enzyme-linked immunosorbent assay. The bioactivity changes of dissolvable immobilized FGF2 were measured by methyl-thiazolyl-tetrazolium assay. Behavior of HGFs was evaluated using adhesion and methyl-thiazolyl-tetrazolium bromide assays. The FGF2 remained for several days on the modified surface on which HGFs were cultured. Over 90% of the dissolvable immobilized FGF2 had been eluted by Day 9, whereas the FGF2 activity was found to diminish gradually from Day 1 to Day 9. The titania nanotubular surface with an optimal preparing concentration (500 ng/mL) of FGF2 immobilization exhibited improved HGF functions such as cellular attachment, proliferation, and extracellular matrix-related gene expression. Moreover, significant bidirectional as well as concentration- and time-dependent bioactivity was observed. Synergism of the FGF2-impregnated titanium dioxide nanotubular surface revealed good gingival-implant integration, indicating that these materials might have promising applications in dentistry and other biomedical devices.

  4. Electric fields control the orientation of peptides irreversibly immobilized on radical-functionalized surfaces.

    Science.gov (United States)

    Martin, Lewis J; Akhavan, Behnam; Bilek, Marcela M M

    2018-01-24

    Surface functionalization of an implantable device with bioactive molecules can overcome adverse biological responses by promoting specific local tissue integration. Bioactive peptides have advantages over larger protein molecules due to their robustness and sterilizability. Their relatively small size presents opportunities to control the peptide orientation on approach to a surface to achieve favourable presentation of bioactive motifs. Here we demonstrate control of the orientation of surface-bound peptides by tuning electric fields at the surface during immobilization. Guided by computational simulations, a peptide with a linear conformation in solution is designed. Electric fields are used to control the peptide approach towards a radical-functionalized surface. Spontaneous, irreversible immobilization is achieved when the peptide makes contact with the surface. Our findings show that control of both peptide orientation and surface concentration is achieved simply by varying the solution pH or by applying an electric field as delivered by a small battery.

  5. Surface engineering of Ti-O films by photochemical immobilization of gelatin

    International Nuclear Information System (INIS)

    Weng, Y.J.; Ren, J.R.; Huang, N.; Wang, J.; Chen, J.Y.; Leng, Y.X.; Liu, H.Q.

    2008-01-01

    Crystalline Ti-O films were prepared by unbalanced magnetron sputtering and the structure was confirmed by XRD. An organic layer of 3-aminopropylphosphonic acid (APP) was first introduced on the Ti-O films by self-assembling. The stability of the APP on Ti-O films was confirmed by XPS and FTIR analysis. Simultaneously, azido group was introduced in gelatin molecule to act as photoreactive point. The derivated gelatin was spin-coated onto the self-assembled layer and immobilized by UV irradiating. Chemical patterned surface was obtained by using a photomask when irradiating and confirmed by sirius red staining and surface profile analysis. Measured by surface profilometer, the thickness of the immobilized gelatin was about 5-20 nm. The adhering of human endothelial EVC304 cells on APP modified surface was enhanced in the cell culture test. Moreover, the adherence and growth of cells were prior on gelatin-immobilized region visually seen on the patterned surface. This result indicated gelatin-immobilized Ti-O surface can serve as a biocompatible biomaterial for endothelialization

  6. Exposure Scenarios and Unit Dose Factors for the Hanford Immobilized Low Activity Tank Waste Performance Assessment

    Energy Technology Data Exchange (ETDEWEB)

    RITTMANN, P.D.

    1999-12-29

    Exposure scenarios are defined to identify potential pathways and combinations of pathways that could lead to radiation exposure from immobilized tank waste. Appropriate data and models are selected to permit calculation of dose factors for each exposure

  7. Co-immobilization of different enzyme activities to non-woven polyester surfaces.

    Science.gov (United States)

    Nouaimi-Bachmann, Meryem; Skilewitsch, Olga; Senhaji-Dachtler, Saida; Bisswanger, Hans

    2007-03-01

    Co-immobilization was applied to combine complementary enzyme reactions. Therefore, trypsin was co-immobilized together with both, lipase and alpha-amylase, onto the surface of non-woven polyester material. The progress of the immobilization reaction was directly monitored by investigating covalent fixation of the enzymes to the polyester flees using (1)H-MAS-NMR. Co-immobilization of the different types of enzymes to the polyester support showed retained enzymatic activity. However, a competition of binding to the support was observed. Increasing amounts of one type of enzyme reduced the degree of immobilization for the other type. In order to investigate the distribution of trypsin and alpha-amylase on the polyester support, the flees was treated with a mixture of rhodamine isothiocyanate labeled with anti-trypsin antibodies and fluorescein isothiocyanate labeled with anti-alpha-amylase antibodies. Using fluorescence microscopy, the co-immobilization was analyzed by selective excitation of both chromophores at 480 and 530 nm, respectively. In addition, fluorescence spectroscopy was applied by direct labeling of trypsin and lipase prior to co-immobilization to the polyester support. A special prism of plexiglass was constructed, which fit into a 10 x 10 mm fluorescence cuvette in that way that a diagonal plane was formed within the cuvette. The non-woven support was fixed in the cuvette and fluorescence spectra were obtained to characterize the amount of different enzymes linked to the support. Using FRET it was demonstrated that a uniform distribution of the various enzyme species was achieved, where the different enzyme activities are bound on the support in close neighborhood to one another.

  8. Covalent immobilization of stem cell factor and stromal derived factor 1α for in vitro culture of hematopoietic progenitor cells.

    Science.gov (United States)

    Cuchiara, Maude L; Horter, Kelsey L; Banda, Omar A; West, Jennifer L

    2013-12-01

    Hematopoietic stem cells (HSCs) are currently utilized in the treatment of blood diseases, but widespread application of HSC therapeutics has been hindered by the limited availability of HSCs. With a better understanding of the HSC microenvironment and the ability to precisely recapitulate its components, we may be able to gain control of HSC behavior. In this work we developed a novel, biomimetic PEG hydrogel material as a substrate for this purpose and tested its potential with an anchorage-independent hematopoietic cell line, 32D clone 3 cells. We immobilized a fibronectin-derived adhesive peptide sequence, RGDS; a cytokine critical in HSC self-renewal, stem cell factor (SCF); and a chemokine important in HSC homing and lodging, stromal derived factor 1α (SDF1α), onto the surfaces of poly(ethylene glycol) (PEG) hydrogels. To evaluate the system's capabilities, we observed the effects of the biomolecules on 32D cell adhesion and morphology. We demonstrated that the incorporation of RGDS onto the surfaces promotes 32D cell adhesion in a dose-dependent fashion. We also observed an additive response in adhesion on surfaces with RGDS in combination with either SCF or SDF1α. In addition, the average cell area increased and circularity decreased on gel surfaces containing immobilized SCF or SDF1α, indicating enhanced cell spreading. By recapitulating aspects of the HSC microenvironment using a PEG hydrogel scaffold, we have shown the ability to control the adhesion and spreading of the 32D cells and demonstrated the potential of the system for the culture of primary hematopoietic cell populations. Copyright © 2013 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

  9. Transglutaminase-mediated protein immobilization to casein nanolayers created on a plastic surface.

    Science.gov (United States)

    Kamiya, Noriho; Doi, Satoshi; Tominaga, Jo; Ichinose, Hirofumi; Goto, Masahiro

    2005-01-01

    An enzymatic method for covalent and site-specific immobilization of recombinant proteins on a plastic surface was explored. Using Escherichia coli alkaline phosphatase (AP) with a specific peptide tag (MKHKGS) genetically incorporated at the N-terminus as a model (NK-AP), microbial transglutaminase (MTG)-mediated protein immobilization was demonstrated. To generate a reactive surface for MTG, a 96-well polystyrene microtiter plate was physically coated with casein, a good MTG substrate. Successful immobilization of recombinant AP to the nanolayer of casein on the surface of the microtiter plate was verified by the detection of enzymatic activity. Since little activity was observed when wild-type AP was used, immobilization of NK-AP was likely directed by the specific peptide tag. When polymeric casein prepared by MTG was used as a matrix on the plate, the loading capacity of AP was increased about 2-fold compared to when casein was used as the matrix. Transglutaminase-mediated site-specific posttranslational modification of proteins offers one way of generating a variety of protein-based solid formulations for biotechnological applications.

  10. Immobilization of α-Chymotrypsin on the Surface of Magnetic/Gold Core/Shell Nanoparticles

    International Nuclear Information System (INIS)

    Ahmadi, M.K.; Vossoughi, M.

    2013-01-01

    Over the last decade, nanoparticles used as protein carriers have opened new avenues for a variety of biomedical applications. The main concern for these applications is changes in biological activity of immobilized proteins due to conformational changes on the surface of the carrier. To evaluate this concern, the preparation and bio catalyst activity of α-chymotrypsin-Fe 3 O 4 Au core/shell nanoparticles were investigated. First, Fe 3 O 4 Au core/shell nanoparticles were synthesized by coprecipitation method and citrate reduction of HAuCl 4 . TEM imaging revealed a core size of 13 ± 3 nm and a shell thickness of 4 ± 1 nm for synthesized nanoparticles. X-ray diffraction (XRD) was used to study the crystalline structure of the nanoparticles. Next, the enzyme was immobilized on the surface of synthesized nanoparticles by covalent bonding of Au shell with thiol and amine groups present in the protein structure (e.g., cysteine and histidine residues). FTIR and fluorescence spectroscopy were utilized to study secondary and tertiary structures of the immobilized enzyme. Results show that the secondary and tertiary structures of the enzyme remain virtually unchanged after immobilization on the nanoparticles surface. However, the bio catalyst activity of the enzyme was reduced by thirty percent, indicating possible conformational changes or active site occlusion

  11. Three-dimensional immobilization of beta-galactosidase on a silicon surface.

    Science.gov (United States)

    Betancor, Lorena; Luckarift, Heather R; Seo, Jae H; Brand, Oliver; Spain, Jim C

    2008-02-01

    Many alternative strategies to immobilize and stabilize enzymes have been investigated in recent years for applications in biosensors. The entrapment of enzymes within silica-based nanospheres formed through silicification reactions provides high loading capacities for enzyme immobilization, resulting in high volumetric activity and enhanced mechanical stability. Here we report a strategy for chemically associating silica nanospheres containing entrapped enzyme to a silicon support. beta-galactosidase from E. coli was used as a model enzyme due to its versatility as a biosensor for lactose. The immobilization strategy resulted in a three-dimensional network of silica attached directly at the silicon surface, providing a significant increase in surface area and a corresponding 3.5-fold increase in enzyme loading compared to enzyme attached directly at the surface. The maximum activity recovered for a silicon square sample of 0.5 x 0.5 cm was 0.045 IU using the direct attachment of the enzyme through glutaraldehyde and 0.16 IU when using silica nanospheres. The immobilized beta-galactosidase prepared by silica deposition was stable and retained more than 80% of its initial activity after 10 days at 24 degrees C. The ability to generate three-dimensional structures with enhanced loading capacity for biosensing molecules offers the potential to substantially amplify biosensor sensitivity. (c) 2007 Wiley Periodicals, Inc.

  12. Analysis of direct immobilized recombinant protein G on a gold surface

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Hyunhee [Department of Chemical and Biomolecular Engineering, Sogang University, Shinsu-dong, Mapo-gu, Seoul 121-742 (Korea, Republic of); Research Institute for Applied Science and Technology, Sogang University , Shinsu-dong, Mapo-gu, Seoul 121-742 (Korea, Republic of); Kang, Da-Yeon [Department of Chemical and Biomolecular Engineering, Sogang University, Shinsu-dong, Mapo-gu, Seoul 121-742 (Korea, Republic of); Goh, Hyun-Jeong [Interdisciplinary Program of Integrated Biotechnology, Sogang University, Shinsu-dong, Mapo-gu, Seoul 121-742 (Korea, Republic of); Oh, Byung-Keun [Department of Chemical and Biomolecular Engineering, Sogang University, Shinsu-dong, Mapo-gu, Seoul 121-742 (Korea, Republic of); Interdisciplinary Program of Integrated Biotechnology, Sogang University, Shinsu-dong, Mapo-gu, Seoul 121-742 (Korea, Republic of); Singh, Ravindra P. [Interdisciplinary Program of Integrated Biotechnology, Sogang University, Shinsu-dong, Mapo-gu, Seoul 121-742 (Korea, Republic of); Oh, Soo-Min [Department of Chemical and Biomolecular Engineering, Sogang University, Shinsu-dong, Mapo-gu, Seoul 121-742 (Korea, Republic of); Research Institute for Applied Science and Technology, Sogang University , Shinsu-dong, Mapo-gu, Seoul 121-742 (Korea, Republic of); Choi, Jeong-Woo [Department of Chemical and Biomolecular Engineering, Sogang University, Shinsu-dong, Mapo-gu, Seoul 121-742 (Korea, Republic of); Interdisciplinary Program of Integrated Biotechnology, Sogang University, Shinsu-dong, Mapo-gu, Seoul 121-742 (Korea, Republic of)], E-mail: jwchoi@sogang.ac.kr

    2008-09-15

    Abstact: For the immobilization of IgG, various techniques such as chemical linker, thiolated protein G methods, and fragmentation of antibodies have been reported [Y.M. Bae, B.K. Oh, W. Lee, W.H. Lee, J.W. Choi, Biosensors Bioelectron. 21 (2005) 103; W. Lee, B.K. Oh, W.H. Lee, J.W. Choi, Colloids Surf. B-Biointerfaces, 40 (2005) 143; A.A. Karyakin, G.V. Presnova, M.Y. Rubtsova, A.M. Egorov, Anal. Chem. 72 (2000) 3805]. Here, we modified the immunoglobulin Fc-binding B-domain of protein G to contain two cysteine residues at its C-terminus by a genetic engineering technique. The resulting recombinant protein, RPGcys, retained IgG-binding activity in the same manner as native protein G. RPGcys was immobilized on a gold surface by strong affinity between thiol of cysteine and gold. The orientations of both IgG layers immobilized on the base recombinant protein Gs were analyzed by fluorescence microscope, atomic force microscope (AFM), and surface plasmon resonance (SPR). Our data revealed that IgG-binding activity of RPGcys on gold surface significantly increased in comparison to wild type of protein G (RPGwild), which was physically adsorbed due to absence of cysteine residue. Immobilization of highly oriented antibodies based on cysteine-modified protein G could be useful for the fabrication of immunosensor systems.

  13. Nature of immobilization surface affects antibody specificity to placental alkaline phosphatase.

    Science.gov (United States)

    Kumar, Mukesh; Khan, Imran; Sinha, Subrata

    2015-01-01

    Retention of native conformation of immobilized protein is essential for various applications including selection and detection of specific recombinant antibodies (scFvs). Placental alkaline phosphatase (PAP), an onco-fetal antigen expressed on the surface of several tumors, was immobilized on supermagnetic particles for selection of recombinant antibodies from a human phage display antibody library. The isolated antibodies were found to be cross-reactive to either of the isozymes of alkaline phosphatase, i.e., bone alkaline phosphatase (BAP) or intestinal alkaline phosphatase (IAP) and could not be used for tumor targeting. A specific anti-PAP monoclonal antibody H17E2 was tested for retention of specificity under these conditions. Binding of the antibody to magnetic beads conjugated IAP and BAP along with PAP and the ability of the two isozymes to inhibit its binding to PAP depicted the loss of isozyme specificity of the antibody. However, the antibody retained its specificity to PAP immobilized on polyvinyl chloride (PVC) surface. Enzyme activity was observed on both surfaces. This demonstrates that nature of immobilization may affect antigen-antibody binding in subtle ways, resulting in alteration of conformation of the epitopes. This may have consequences for determining the specificity of antibody binding for proteins that share a high degree of homology.

  14. Antimicrobial membrane surfaces via efficient polyethyleneimine immobilization and cationization

    Science.gov (United States)

    Qiu, Wen-Ze; Zhao, Zi-Shu; Du, Yong; Hu, Meng-Xin; Xu, Zhi-Kang

    2017-12-01

    Biofouling control is a major task in membrane separation processes for water treatment and biomedical applications. In this work, N-alkylated polyethylenimine (PEI) is facilely and efficiently introduced onto the membrane surfaces via the co-deposition of catechol (CCh) and PEI, followed by further grafting of PEIs (600 Da, 70 kDa and 750 kDa) and cationization with methyl iodide (CH3I). The physical and chemical properties of the constructed membrane surfaces are characterized with scanning electron microscopy, Fourier transform infrared spectroscopy, X-ray photoelectron spectroscopy, zeta potential and water contact angle measurements. Antibacterial assay reveals that the optimized membrane surfaces possess around 95% antibacterial efficiency against Gram-positive Staphylococcus aureus (S. aureus) with weak adhesion of bacteria cells after 24 h of bacterial contact. Additionally, the membrane surfaces also exhibit much enhanced antifouling property during the filtration of opposite charged bovine serum albumin (BSA). These results demonstrate a useful strategy for the surface modification of separation membranes by a kind of antimicrobial and antifouling coating.

  15. Use of activated carbon as a support medium for H2S biofiltration and effect of bacterial immobilization on available pore surface.

    Science.gov (United States)

    Ng, Y L; Yan, R; Chen, X G; Geng, A L; Gould, W D; Liang, D T; Koe, L C C

    2004-12-01

    The use of support media for the immobilization of microorganisms is widely known to provide a surface for microbial growth and a shelter that protects the microorganisms from inhibitory compounds. In this study, activated carbon is used as a support medium for the immobilization of microorganisms enriched from municipal sewage activated sludge to remove gas-phase hydrogen sulfide (H2S), a major odorous component of waste gas from sewage treatment plants. A series of designed experiments is used to examine the effect on bacteria-immobilized activated carbon (termed "biocarbon") due to physical adsorption, chemical reaction, and microbial degradation in the overall removal of H2S. H2S breakthrough tests are conducted with various samples, including microbe-immobilized carbon and Teflon discs, salts-medium-washed carbon, and ultra-pure water-washed carbon. The results show a higher removal capacity for the microbe-immobilized activated carbon compared with the activated carbon control in a batch biofilter column. The increase in removal capacity is attributed to the role played by the immobilized microorganisms in metabolizing adsorbed sulfur and sulfur compounds on the biocarbon, hence releasing the adsorption sites for further H2S uptake. The advantage for activated carbon serving as the support medium is to adsorb a high initial concentration of substrate and progressively release this for microbial degradation, hence acting as a buffer for the microorganisms. Results obtained from surface area and pore size distribution analyses of the biocarbon show a correlation between the available surface area and pore volume with the extent of microbial immobilization and H2S uptake. The depletion of surface area and pore volume is seen as one of the factors which cause the onset of column breakthrough. Microbial growth retardation is due to the accumulation of metabolic products (i.e., sulfuric acid); and a lack of water and nutrient salts in the batch biofilter are other

  16. Immobilization of carbon nanotubes and metallophthalocyanines on conductive surfaces by electrochemical means for electroanalytical purposes

    Energy Technology Data Exchange (ETDEWEB)

    Porras Gutierrez, A.; Gutierrez Granados, S. [Centre national de la recherche scientifique, Paris (France). Unite de Pharmacologie Chimique et Genetique; Guanajuato Univ. Guanajuato (Mexico). Inst. de Investigaciones Cientificas; Richard, C.; Griveau, S.; Bedioui, F. [Centre national de la recherche scientifique, Paris (France). Unite de Pharmacologie Chimique et Genetique; Zagal, J.H. [Santiago Univ. de Chile, Santiago (Chile)

    2008-07-01

    Carbon nanotubes (CNT) have been touted as viable candidates for the design of new electrode materials because of their high conductivity and high specific surface area. This study explored the use of electrochemical methods to immobilize single walled carbon nanotubes (SWCNT) on glassy carbon (GC) in a stable and controlled fashion. Two electrochemical routes were investigated to get the stable immobilization of nanotubes, notably (1) electropolymerization of conducting polymers in presence of SWCNT, and (2) the electrochemical grafting of diazonium salts in presence of SWCNT. The objective was to obtain chemically and mechanically stable composite GC/SWCNT electrodes. The electrochemical performances and reactivity of the electrodes were analyzed by voltammetry and by scanning electrochemical microscopy. The optimized immobilization methods were then applied to the conception of electrocatalysts hybrids, by co-immobilization of nanotubes with well-known redox catalyst metallocomplexes for activation of the electro-oxidation of biologically relevant thiol. The study showed that the nanocomposite material based on the combined use of metallophthalocynines, functionalized SWCNTs and electropolymerizable matrices enables the assembly of highly stable electrodes with better electrocatalytic oxidation of thiols. This fast procedure to modify glassy carbon (GC) electrode using commercially available cobalt phthalocyanine (CoPc) and tetrasulfonated nickel phthalocyanine (NiTSPc), oxidized single walled carbon nanotubes SWCNT and electropolymerized polypyrrole or diazonium derivatives. It was concluded that the electrodes are highly stable and the tailored hybrid surfaces improves electron transfer. 4 refs.

  17. Factors affecting the immobilization of fungal biomass on CNT as a biosorbent for textile dyes removal

    Science.gov (United States)

    Adebayo Bello, Ibrahim; Kabbashi, Nassereldeen A.; Zahangir Alam, Md; Alkhatib, Ma'an F.; Nabilah Murad, Fatin

    2017-07-01

    Effluents from dye and textile industries are highly contaminated and toxic to the environment. High concentration of non-biodegradable compounds contributes to increased biochemical oxygen demand (BOD) and chemical oxygen demand (COD) of the wastewater bodies. Dyes found in wastewater from textile industries are carcinogenic, mutagenic or teratogenic. Biological processes involving certain bacteria, fungi and activated carbon have been employed in treating wastewater. These methods are either inefficient or ineffective. These complexities necessitates search for new approaches that will offset all the shortcomings of the present solutions to the challenges faced with textile wastewater management. This study produced a new biosorbent by the immobilization of fungal biomass on carbon nanotubes. The new biosorbent is called “carbon nanotubes immobilized biomass (CNTIB)” which was produced by immobilization technique. A potential fungal strain, Aspergillus niger was selected on the basis of biomass production. It was found out in this studies that fungal biomass were better produced in acidic medium. Aspergillus niger was immobilized on carbon nanotubes. One-factor-at-a time (OFAT) was employed to determine the effect of different factors on the immobilization of fungal biomass on carbon nanotubes and optimum levels at which the three selected parameters (pH, culture time and agitation rate) would perform. Findings from OFAT showed that the optimum conditions for immobilization are a pH of 5, agitation rate of 150rpm and a culture time of 5 days.

  18. Controlled preparation of thin fibrin films immobilized at solid surfaces

    Czech Academy of Sciences Publication Activity Database

    Riedel, Tomáš; Brynda, Eduard; Dyr, J. E.; Houska, Milan

    88A, č. 2 (2009), s. 437-447 ISSN 1549-3296 R&D Projects: GA AV ČR(CZ) IAA400500507 Institutional research plan: CEZ:AV0Z40500505 Keywords : fibrin gel coatings * thin films * tissue engineering Subject RIV: CE - Biochemistry Impact factor: 2.816, year: 2009

  19. Surface modification of chitosan/PEO nanofibers by air dielectric barrier discharge plasma for acetylcholinesterase immobilization

    Energy Technology Data Exchange (ETDEWEB)

    Dorraki, Naghme, E-mail: n.dorraki@web.sbu.ac.ir [Laser and Plasma Research Institute, Shahid Beheshti University, Evin 1983963113, Tehran (Iran, Islamic Republic of); Safa, Nasrin Navab [Laser and Plasma Research Institute, Shahid Beheshti University, Evin 1983963113, Tehran (Iran, Islamic Republic of); Jahanfar, Mehdi [Protein Research Center, Shahid Beheshti University, Evin 1983963113, Tehran (Iran, Islamic Republic of); Ghomi, Hamid [Laser and Plasma Research Institute, Shahid Beheshti University, Evin 1983963113, Tehran (Iran, Islamic Republic of); Ranaei-Siadat, Seyed-Omid [Protein Research Center, Shahid Beheshti University, Evin 1983963113, Tehran (Iran, Islamic Republic of)

    2015-09-15

    Highlights: • We used an economical and effective method for surface modification. • Chitosan/PEO nanofibrous membranes were modified by air-DBD plasma. • The most NH{sub 3}{sup +} group was generated on the 6 min plasma modified membrane. • We immobilized acetylcholinesterase on the plasma modified and unmodified membranes. • More enzyme activity was detected on the modified membrane by plasma. - Abstract: There are different methods to modify polymer surfaces for biological applications. In this work we have introduced air-dielectric barrier discharge (DBD) plasma at atmospheric pressure as an economical and safe method for modifying the surface of electrospun chitosan/PEO (90/10) nanofibers for acetylcholinesterase (AChE) immobilization. According to the contact angle measurement results, the nanofibers become highly hydrophilic when they are exposed to the DBD plasma for 6 min in compared to unmodified membrane. Attenuated total reflectance-Fourier transform infrared spectroscopy (ATR-FTIR) results reveal hydroxyl, C=O and NH{sub 3}{sup +} polar groups increment after 6 min plasma treatment. Contact angle measurements and ATR-FTIR results are confirmed by X-ray photoelectron spectroscopy (XPS). AChE at pH 7.4 carries a negative charge and after immobilization on the surface of plasma-treated nanofibrous membrane attracts the NH{sub 3}{sup +} group and more enzyme activity is detected on the plasma-modified nanofibers for 6 min in compared to unmodified nanofibers. Atomic force microscopy (AFM) and scanning electron microscopy (SEM) are used for the surface topography and morphology characterization. The results have proved that air-DBD plasma is a suitable method for chitosan/PEO nanofibrous membrane modification as a biodegradable and functionalized substrate for enzyme immobilization.

  20. Surface dosimetry for breast radiotherapy in the presence of immobilization cast material

    International Nuclear Information System (INIS)

    Kelly, Andrew; Hardcastle, Nicholas; Metcalfe, Peter; Cutajar, Dean; Quinn, Alexandra; Cardoso, Michael; Rosenfeld, Anatoly; Foo, Kerwyn; Barlin, Sheree

    2011-01-01

    Curative breast radiotherapy typically leaves patients with varying degrees of cosmetic damage. One problem interfering with cosmetically acceptable breast radiotherapy is the external contour for large pendulous breasts which often results in high doses to skin folds. Thermoplastic casts are often employed to secure the breasts to maintain setup reproducibility and limit the presence of skin folds. This paper aims to determine changes in surface dose that can be attributed to the use of thermoplastic immobilization casts. Skin dose for a clinical hybrid conformal/IMRT breast plan was measured using radiochromic film and MOSFET detectors at a range of water equivalent depths representative of the different skin layers. The radiochromic film was used as an integrating dosimeter, while the MOSFETs were used for real-time dosimetry to isolate the contribution of skin dose from individual IMRT segments. Strips of film were placed at various locations on the breast and the MOSFETs were used to measure skin dose at 16 positions spaced along the film strips for comparison of data. The results showed an increase in skin dose in the presence of the immobilization cast of up to 45.7% and 62.3% of the skin dose without the immobilization cast present as measured with Gafchromic EBT film and MOSFETs, respectively. The increase in skin dose due to the immobilization cast varied with the angle of beam incidence and was greatest when the beam was normally incident on the phantom. The increase in surface dose with the immobilization cast was greater under entrance dose conditions compared to exit dose conditions.

  1. Surface dosimetry for breast radiotherapy in the presence of immobilization cast material

    Energy Technology Data Exchange (ETDEWEB)

    Kelly, Andrew; Hardcastle, Nicholas; Metcalfe, Peter; Cutajar, Dean; Quinn, Alexandra; Cardoso, Michael; Rosenfeld, Anatoly [Centre for Medical Radiation Physics, University of Wollongong, Wollongong, NSW (Australia); Foo, Kerwyn [Sydney Medical School, University of Sydney, Sydney, NSW (Australia); Barlin, Sheree, E-mail: anatoly@uow.edu.au [Illawarra Cancer Care Centre, Wollongong Hospital, Wollongong, NSW (Australia)

    2011-02-21

    Curative breast radiotherapy typically leaves patients with varying degrees of cosmetic damage. One problem interfering with cosmetically acceptable breast radiotherapy is the external contour for large pendulous breasts which often results in high doses to skin folds. Thermoplastic casts are often employed to secure the breasts to maintain setup reproducibility and limit the presence of skin folds. This paper aims to determine changes in surface dose that can be attributed to the use of thermoplastic immobilization casts. Skin dose for a clinical hybrid conformal/IMRT breast plan was measured using radiochromic film and MOSFET detectors at a range of water equivalent depths representative of the different skin layers. The radiochromic film was used as an integrating dosimeter, while the MOSFETs were used for real-time dosimetry to isolate the contribution of skin dose from individual IMRT segments. Strips of film were placed at various locations on the breast and the MOSFETs were used to measure skin dose at 16 positions spaced along the film strips for comparison of data. The results showed an increase in skin dose in the presence of the immobilization cast of up to 45.7% and 62.3% of the skin dose without the immobilization cast present as measured with Gafchromic EBT film and MOSFETs, respectively. The increase in skin dose due to the immobilization cast varied with the angle of beam incidence and was greatest when the beam was normally incident on the phantom. The increase in surface dose with the immobilization cast was greater under entrance dose conditions compared to exit dose conditions.

  2. Surface dosimetry for breast radiotherapy in the presence of immobilization cast material

    Science.gov (United States)

    Kelly, Andrew; Hardcastle, Nicholas; Metcalfe, Peter; Cutajar, Dean; Quinn, Alexandra; Foo, Kerwyn; Cardoso, Michael; Barlin, Sheree; Rosenfeld, Anatoly

    2011-02-01

    Curative breast radiotherapy typically leaves patients with varying degrees of cosmetic damage. One problem interfering with cosmetically acceptable breast radiotherapy is the external contour for large pendulous breasts which often results in high doses to skin folds. Thermoplastic casts are often employed to secure the breasts to maintain setup reproducibility and limit the presence of skin folds. This paper aims to determine changes in surface dose that can be attributed to the use of thermoplastic immobilization casts. Skin dose for a clinical hybrid conformal/IMRT breast plan was measured using radiochromic film and MOSFET detectors at a range of water equivalent depths representative of the different skin layers. The radiochromic film was used as an integrating dosimeter, while the MOSFETs were used for real-time dosimetry to isolate the contribution of skin dose from individual IMRT segments. Strips of film were placed at various locations on the breast and the MOSFETs were used to measure skin dose at 16 positions spaced along the film strips for comparison of data. The results showed an increase in skin dose in the presence of the immobilization cast of up to 45.7% and 62.3% of the skin dose without the immobilization cast present as measured with Gafchromic EBT film and MOSFETs, respectively. The increase in skin dose due to the immobilization cast varied with the angle of beam incidence and was greatest when the beam was normally incident on the phantom. The increase in surface dose with the immobilization cast was greater under entrance dose conditions compared to exit dose conditions.

  3. Analysis of the surface density and reactivity of perfluorophenylazide and the impact on ligand immobilization

    Energy Technology Data Exchange (ETDEWEB)

    Zorn, Gilad, E-mail: zorn@ge.com; Castner, David G. [National ESCA and Surface Analysis Center for Biomedical Problems, Departments of Bioengineering and Chemical Engineering, University of Washington, Box 351653, Seattle, Washington 98195-1653 (United States); Tyagi, Anuradha; Wang, Xin; Wang, Hui; Yan, Mingdi, E-mail: Mingdi-Yan@uml.edu [Department of Chemistry, Portland State University, Portland, Oregon 97207-0751 (United States)

    2015-03-15

    Perfluorophenylazide (PFPA) chemistry is a novel method for tailoring the surface properties of solid surfaces and nanoparticles. It is general and versatile, and has proven to be an efficient way to immobilize graphene, proteins, carbohydrates, and synthetic polymers. The main thrust of this work is to provide a detailed investigation on the chemical composition and surface density of the PFPA tailored surface. Specifically, gold surfaces were treated with PFPA-derivatized (11-mercaptoundecyl)tetra(ethylene glycol) (PFPA-MUTEG) mixed with 2-[2-(2-mercaptoethoxy)ethoxy]ethanol (MDEG) at varying solution mole ratios. Complementary analytical techniques were employed to characterize the resulting films including Fourier transform infrared spectroscopy to detect fingerprints of the PFPA group, x-ray photoelectron spectroscopy and ellipsometry to study the homogeneity and uniformity of the films, and near edge x-ray absorption fine structures to study the electronic and chemical structure of the PFPA groups. Results from these studies show that the films prepared from 90:10 and 80:20 PFPA-MUTEG/MDEG mixed solutions exhibited the highest surface density of PFPA and the most homogeneous coverage on the surface. A functional assay using surface plasmon resonance with carbohydrates covalently immobilized onto the PFPA-modified surfaces showed the highest binding affinity for lectin on the PFPA-MUTEG/MDEG film prepared from a 90:10 solution.

  4. Analysis of the surface density and reactivity of perfluorophenylazide and the impact on ligand immobilization

    International Nuclear Information System (INIS)

    Zorn, Gilad; Castner, David G.; Tyagi, Anuradha; Wang, Xin; Wang, Hui; Yan, Mingdi

    2015-01-01

    Perfluorophenylazide (PFPA) chemistry is a novel method for tailoring the surface properties of solid surfaces and nanoparticles. It is general and versatile, and has proven to be an efficient way to immobilize graphene, proteins, carbohydrates, and synthetic polymers. The main thrust of this work is to provide a detailed investigation on the chemical composition and surface density of the PFPA tailored surface. Specifically, gold surfaces were treated with PFPA-derivatized (11-mercaptoundecyl)tetra(ethylene glycol) (PFPA-MUTEG) mixed with 2-[2-(2-mercaptoethoxy)ethoxy]ethanol (MDEG) at varying solution mole ratios. Complementary analytical techniques were employed to characterize the resulting films including Fourier transform infrared spectroscopy to detect fingerprints of the PFPA group, x-ray photoelectron spectroscopy and ellipsometry to study the homogeneity and uniformity of the films, and near edge x-ray absorption fine structures to study the electronic and chemical structure of the PFPA groups. Results from these studies show that the films prepared from 90:10 and 80:20 PFPA-MUTEG/MDEG mixed solutions exhibited the highest surface density of PFPA and the most homogeneous coverage on the surface. A functional assay using surface plasmon resonance with carbohydrates covalently immobilized onto the PFPA-modified surfaces showed the highest binding affinity for lectin on the PFPA-MUTEG/MDEG film prepared from a 90:10 solution

  5. Detonation nanodiamonds biofunctionalization and immobilization to titanium alloy surfaces as first steps towards medical application

    Directory of Open Access Journals (Sweden)

    Juliana P. L. Gonçalves

    2014-11-01

    Full Text Available Due to their outstanding properties nanodiamonds are a promising nanoscale material in various applications such as microelectronics, polishing, optical monitoring, medicine and biotechnology. Beyond the typical diamond characteristics like extreme hardness or high thermal conductivity, they have additional benefits as intrinsic fluorescence due to lattice defects without photobleaching, obtained during the high pressure high temperature process. Further the carbon surface and its various functional groups in consequence of the synthesis, facilitate additional chemical and biological modification. In this work we present our recent results on chemical modification of the nanodiamond surface with phosphate groups and their electrochemically assisted immobilization on titanium-based materials to increase adhesion at biomaterial surfaces. The starting material is detonation nanodiamond, which exhibits a heterogeneous surface due to the functional groups resulting from the nitrogen-rich explosives and the subsequent purification steps after detonation synthesis. Nanodiamond surfaces are chemically homogenized before proceeding with further functionalization. Suspensions of resulting surface-modified nanodiamonds are applied to the titanium alloy surfaces and the nanodiamonds subsequently fixed by electrochemical immobilization. Titanium and its alloys have been widely used in bone and dental implants for being a metal that is biocompatible with body tissues and able to bind with adjacent bone during healing. In order to improve titanium material properties towards biomedical applications the authors aim to increase adhesion to bone material by incorporating nanodiamonds into the implant surface, namely the anodically grown titanium dioxide layer. Differently functionalized nanodiamonds are characterized by infrared spectroscopy and the modified titanium alloys surfaces by scanning and transmission electron microscopy. The process described shows an

  6. The Effect of Surface Functionalization on the Immobilization of Gold Nanoparticles on Graphene Sheets

    Directory of Open Access Journals (Sweden)

    Min Song

    2012-01-01

    Full Text Available In our study, graphene oxide is synthesized by Hummers method. And then, carboxylic acid functionalized graphene (graphene-COOH, thiol-functionalized graphene (graphene-SH, and highly dispersive graphene are prepared by chemical modification of respective groups on the graphene surface. Furthermore, we explore a solution-based approach to prepare three differently functionalized graphene-gold composites by one-step chemical reduction of AuCl4 - ions in respective functionalized graphene suspensions, where the gold nanoparticles are deposited on the functionalized graphene surface during their synthesis process. In addition, we compare the influence of surface functionalization on the growth of gold nanoparticles on graphene surface. Transmission electron morphology (TEM and ultraviolet-visible (UV-Vis spectroscopy are employed to study the effect of surface functionalities on AuNPs distribution onto the graphene surface and demonstrate the successful immobilization of AuNPs on graphene surface.

  7. Protein immobilization on epoxy-activated thin polymer films: effect of surface wettability and enzyme loading.

    Science.gov (United States)

    Chen, Bo; Pernodet, Nadine; Rafailovich, Miriam H; Bakhtina, Asya; Gross, Richard A

    2008-12-02

    A series of epoxy-activated polymer films composed of poly(glycidyl methacrylate/butyl methacrylate/hydroxyethyl methacrylate) were prepared. Variation in comonomer composition allowed exploration of relationships between surface wettability and Candida antartica lipase B (CALB) binding to surfaces. By changing solvents and polymer concentrations, suitable conditions were developed for preparation by spin-coating of uniform thin films. Film roughness determined by AFM after incubation in PBS buffer for 2 days was less than 1 nm. The occurrence of single CALB molecules and CALB aggregates at surfaces was determined by AFM imaging and measurements of volume. Absolute numbers of protein monomers and multimers at surfaces were used to determine values of CALB specific activity. Increased film wettability, as the water contact angle of films increased from 420 to 550, resulted in a decreased total number of immobilized CALB molecules. With further increases in the water contact angle of films from 55 degrees to 63 degrees, there was an increased tendency of CALB molecules to form aggregates on surfaces. On all flat surfaces, two height populations, differing by more than 30%, were observed from height distribution curves. They are attributed to changes in protein conformation and/or orientation caused by protein-surface and protein-protein interactions. The fraction of molecules in these populations changed as a function of film water contact angle. The enzyme activity of immobilized films was determined by measuring CALB-catalyzed hydrolysis of p-nitrophenyl butyrate. Total enzyme specific activity decreased by decreasing film hydrophobicity.

  8. Surface functionalization of superparamagnetic nanoparticles encapsulated by chitosan for protein immobilization

    International Nuclear Information System (INIS)

    Sousa, Jose Silva de

    2010-01-01

    Nanoscience and nanotechnology have opened up numerous developments of devices and systems on the nanometer scale, with new molecular organization, properties and functions. In this context, the polymeric magnetic nanoparticles are composites formed by magnetic materials with a particle size between 1 and 100 nm combined with functional polymers. They are well-known and have been widely studied because of its applications in various technology areas. Applications on the biological and medical areas include separation and immobilization of enzymes and proteins, improved techniques of magnetic resonance imaging and diagnostic systems for controlled drug delivery. In this work, proteins were immobilized on the surface of a biopolymer combined with superparamagnetic particles of magnetite. The biopolymer chitosan was used, cross-linked and functionalized with glutaraldehyde, applicable to the biological assays. Three types of magnetic composites were obtained, which were called QM1Glu, QM2NaGlu and QM3Glu. They were characterized by X-ray diffraction, scanning electron microscopy, vibrating sample magnetometry, differential scanning calorimetry, thermogravimetry and infrared spectroscopy. They were evaluated concerning the immobilization of the proteins bovine serum albumin (BSA), collagen and trypsin. The study showed that the immobilization of proteins on the biopolymer occurred in 30 min of incubation. The magnetic composite of non functionalized chitosan (QM3) was also evaluated. For trypsin, it was found that the immobilization potential of QM3 was higher than that observed for QM3Glu. After 30 days, the trypsin of the QM3-Trip and QM3Glu-Trip was still with activity. The activity and the enzyme kinetics of the QM3Glu-Trip with the substrate BApNA were demonstrated. (author)

  9. Enhancing anticoagulation and endothelial cell proliferation of titanium surface by sequential immobilization of poly(ethylene glycol) and collagen

    International Nuclear Information System (INIS)

    Pan, Chang-Jiang; Hou, Yan-Hua; Ding, Hong-Yan; Dong, Yun-Xiao

    2013-01-01

    In the present study, poly(ethylene glycol) (PEG) and collagen I were sequentially immobilized on the titanium surface to simultaneously improve the anticoagulation and endothelial cell proliferation. Attenuated total reflectance Fourier transform infrared spectroscopy (ATR-FTIR) and X-ray photoelectron spectroscopy analysis confirmed that PEG and collagen I were successfully immobilized on the titanium surface. Water contact angle results suggested the excellent hydrophilic surface after the immobilization. The anticoagulation experiments demonstrated that the immobilized PEG and collagen I on the titanium surface could not only obviously prevent platelet adhesion and aggregation but also prolong activated partial thromboplastin time (APTT), leading to the improved blood compatibility. Furthermore, immobilization of collagen to the end of PEG chain did not abate the anticoagulation. As compared to those on the pristine and PEG-modified titanium surfaces, endothelial cells exhibited improved proliferative profiles on the surface modified by the sequential immobilization of PEG and collagen in terms of CCK-8 assay, implying that the modified titanium may promote endothelialization without abating the blood compatibility. Our method may be used to modify the surface of blood-contacting biomaterials such as titanium to promote endothelialization and improve the anticoagulation, it may be helpful for development of the biomedical devices such as coronary stents, where endothelializaton and excellent anticoagulation are required.

  10. Preservation of surface-dependent properties of viral antigens following immobilization on particulate ceramic delivery vehicles.

    Science.gov (United States)

    Kossovsky, N; Gelman, A; Sponsler, E; Rajguru, S; Torres, M; Mena, E; Ly, K; Festekjian, A

    1995-05-01

    B-cell stimulation for the purpose of evoking an effective neutralizing humoral immune response is a surface phenomenon that is exquisitely specific to antigen conformation. Consequently, successful delivery of antigen, such as would be desired in a vaccine, entails preservation of an antigen's apparent native surface (conformational) properties. Prior to testing the actual vaccinating efficacy of delivered antigens, the surface properties could be assessed through a variety of in vitro and in vivo assays in which the measurement standard would be the properties of the antigens in their native state (whole virus). Using surface modified nanocrystalline carbon and calcium-phosphate ceramic particulates (carbon ceramics and brushite), we evaluated the surface activity of immobilized non-nuclear material extracted from HIV-1. Physical characterization showed that the particles with immobilized antigen ("HIV decoys") measured 50 nm in diameter (HIV = 50-100 nm) and exhibited the same zeta potentials as whole (live) HIV. In vitro testing showed that the HIV decoys were recognized by both conformationally nonspecific and specific monoclonal antibodies, were recognized by human IgG from HIV antibody-positive patients, and could promote surface agglomeration among malignant T-cells similar to live HIV. Last, in vivo testing in three vaccinated animal species showed that the HIV decoys elicited humoral and cellular immune responses similar to that evoked by whole (live) HIV.

  11. Methods for generation of reporter phages and immobilization of active bacteriophages on a polymer surface

    Science.gov (United States)

    Applegate, Bruce Michael (Inventor); Perry, Lynda Louise (Inventor); Morgan, Mark Thomas (Inventor); Kothapalli, Aparna (Inventor)

    2012-01-01

    Novel reporter bacteriophages are provided. Provided are compositions and methods that allow bacteriophages that are used for specific detection or killing of E. coli 0157:H7 to be propagated in nonpathogenic E. coli, thereby eliminating the safety and security risks of propagation in E. coli 0157:H7. Provided are compositions and methods for attaching active bacteriophages to the surface of a polymer in order to kill target bacteria with which the phage comes into contact. Provided are modified bacteriophages immobilized to a surface, which capture E. coli 0157:H7 and cause the captured cells to emit light or fluorescence, allowing detection of the bacteria in a sample.

  12. High density gold nanoparticles immobilized on surface via plasma deposited APTES film for decomposing organic compounds in microchannels

    Science.gov (United States)

    Rao, Xi; Guyon, Cédric; Ognier, Stephanie; Da Silva, Bradley; Chu, Chenglin; Tatoulian, Michaël; Hassan, Ali Abou

    2018-05-01

    Immobilization of colloidal particles (e.g. gold nanoparticles (AuNps)) on the inner surface of micro-/nano- channels has received a great interest for catalysis. A novel catalytic ozonation setup using a gold-immobilized microchannel reactor was developed in this work. To anchor AuNps, (3-aminopropyl) triethoxysilane (APTES) with functional amine groups was deposited using plasma enhanced chemical vapor deposition (PECVD) process. The results clearly evidenced that PECVD processing exhibited relatively high efficiency for grafting amine groups and further immobilizing AuNPs. The catalytic activity of gold immobilized microchannel was evaluated by pyruvic acid ozonation. The decomposition rate calculated from High Performance Liquid Chromatography (HPLC) indicated a much better catalytic performance of gold in microchannel than that in batch. The results confirmed immobilizing gold nanoparticles on plasma deposited APTES for preparing catalytic microreactors is promising for the wastewater treatment in the future.

  13. Immobilization of β-Galactosidase onto Functionalized Graphene Nano-sheets Using Response Surface Methodology and Its Analytical Applications

    Science.gov (United States)

    Kishore, Devesh; Talat, Mahe; Srivastava, Onkar Nath; Kayastha, Arvind M.

    2012-01-01

    Background β-Galactosidase is a vital enzyme with diverse application in molecular biology and industries. It was covalently attached onto functionalized graphene nano-sheets for various analytical applications based on lactose reduction. Methodology/Principal Findings Response surface methodology based on Box-Behnken design of experiment was used for determination of optimal immobilization conditions, which resulted in 84.2% immobilization efficiency. Native and immobilized functionalized graphene was characterized with the help of transmission and scanning electron microscopy, followed by Fourier transform infrared (FTIR) spectroscopy. Functionalized graphene sheets decorated with islands of immobilized enzyme were evidently visualized under both transmission and scanning electron microscopy after immobilization. FTIR spectra provided insight on various chemical interactions and bonding, involved during and after immobilization. Optimum temperature and energy of activation (Ea) remains unchanged whereas optimum pH and Km were changed after immobilization. Increased thermal stability of enzyme was observed after conjugating the enzyme with functionalized graphene. Significance Immobilized β-galactosidase showed excellent reusability with a retention of more than 92% enzymatic activity after 10 reuses and an ideal performance at broad ranges of industrial environment. PMID:22815797

  14. Immobilization mechanisms of deoxyribonucleic acid (DNA) to hafnium dioxide (HfO2) surfaces for biosensing applications.

    Science.gov (United States)

    Fahrenkopf, Nicholas M; Rice, P Zachary; Bergkvist, Magnus; Deskins, N Aaron; Cady, Nathaniel C

    2012-10-24

    Immobilization of biomolecular probes to the sensing substrate is a critical step for biosensor fabrication. In this work we investigated the phosphate-dependent, oriented immobilization of DNA to hafnium dioxide surfaces for biosensing applications. Phosphate-dependent immobilization was confirmed on a wide range of hafnium oxide surfaces; however, a second interaction mode was observed on monoclinic hafnium dioxide. On the basis of previous materials studies on these films, DNA immobilization studies, and density functional theory (DFT) modeling, we propose that this secondary interaction is between the exposed nucleobases of single stranded DNA and the surface. The lattice spacing of monoclinic hafnium dioxide matches the base-to-base pitch of DNA. Monoclinic hafnium dioxide is advantageous for nanoelectronic applications, yet because of this secondary DNA immobilization mechanism, it could impede DNA hybridization or cause nonspecific surface intereactions. Nonetheless, DNA immobilization on polycrystalline and amorphous hafnium dioxide is predominately mediated by the terminal phosphate in an oriented manner which is desirable for biosensing applications.

  15. Surface Modification of Polypropylene Microporous Membrane by Atmospheric-Pressure Plasma Immobilization of N,N-dimethylamino Ethyl Methacrylate

    International Nuclear Information System (INIS)

    Zhong Shaofeng

    2010-01-01

    Surface modification of polypropylene microporous membrane (PPMM) was performed by atmospheric pressure dielectric barrier discharge plasma immobilization of N,N-dimethylamino ethyl methacrylate (DMAEMA). Structural and morphological changes on the membrane surface were characterized by attenuated total reflection-Fourier transform infrared spectroscopy (FT-IR/ATR), X-ray photoelectron spectroscope (XPS) and field emission scanning electron microscopy (FE-SEM). Water contact angles of the membrane surfaces were also measured by the sessile drop method. Results reveal that both the plasma-treating conditions and the adsorbed DMAEMA amount have remarkable effects on the immobilization degree of DMAEMA. Peroxide determination by 1,1-diphenyl-2-picrvlhydrazyl (DPPH) method verifies the exsistence of radicals induced by plasma, which activize the immobilization reaction. Pure water contact angle on the membrane surface decreased with the increase of DMAEMA immobilization degree, which indicates an enhanced hydrophilicity for the modified membranes. The effects of immobilization degrees on pure water fluxes were also measured. It is shown that pure water fluxes first increased with immobilization degree and then decreased. Finally, permeation of bovine serum albumin (BSA) and lysozyme solution were measured to evaluate the antifouling property of the DMAEMA-modified membranes, from which it is shown that both hydrophilicity and electrostatic repulsion are beneficial for membrane antifouling.

  16. Strategies For Immobilization Of Bioactive Organic Molecules On Titanium Implant Surfaces – A Review

    Directory of Open Access Journals (Sweden)

    Panayotov Ivan V.

    2015-03-01

    Full Text Available Numerous approaches have been used to improve the tissue-implant interface of titanium (Ti and titanium alloy (Ti6Al4V. They all aim at increasing cell migration and attachment to the metal, preventing unspecific protein adsorption and improving post-implantation healing process. Promising methods for titanium and titanium alloy surface modification are based on the immobilization of biologically active organic molecules. New and interesting biochemical approaches to such surface modification include layer-by-layer deposition of polyelectrolyte films, phage display-selected surface binding peptides and self-assembled DNA monolayer systems. The present review summarizes the scientific information about these methods, which are at in vitro or in vivo development stages, and hopes to promote their future application in dental implantology and in oral and maxillofacial surgery.

  17. Soluble and immobilized graphene oxide activates complement system differently dependent on surface oxidation state.

    Science.gov (United States)

    Wibroe, Peter P; Petersen, Søren V; Bovet, Nicolas; Laursen, Bo W; Moghimi, S Moein

    2016-02-01

    Graphene oxide (GO) is believed to become applicable in biomedical products and medicine, thereby necessitating appropriate safety evaluation dependent on their applications and the route of administration. We have examined the effect of GO form (in solution versus immobilized) and oxidation state on two related elements of innate immunity: the complement system and interleukin-6 (IL-6) release in human blood. In solution, there was a decrease in GO-mediated complement activation with decreasing surface oxygen content (and altered oxygen functionality), whereas with immobilized GO complement response were reversed and increased with decreasing oxygen content. GO solutions, at concentrations below complement activating threshold, did not induce IL-6 release from human blood leukocytes, and further dampened lipopolysaccharide-induced IL-6 release in the whole blood. The latter effect became more profound with GO's having higher oxygen content. This protective role of GO solutions, however, disappeared at higher concentrations above complement-activating threshold. We discuss these results in relation to GO surface structure and properties, and implications for local administration and development of GO-based implantable devices. Copyright © 2015 Elsevier Ltd. All rights reserved.

  18. Combining Surface Analytical and Computational Techniques to Investigate Orientation Effects of Immobilized Proteins

    Science.gov (United States)

    Harrison, Elisa Turla

    Controlling how proteins are immobilized (e.g. controlling their orientation and conformation) is essential for developing and optimizing the performance of in vitro protein-binding devices, such as enzyme-linked immunosorbent assays. The objective of this work is to develop new methodologies to study proteins and complex mixtures of proteins immobilized onto surfaces. The focus of this study was to control and characterize the orientation of protein G B1, an IgG antibody-binding domain of protein G, on well-defined surfaces as well as measure the effect of protein G B1 orientation on IgG antibody binding using a variety of surface analytical and computational techniques. The surface sensitivity of time-of-flight secondary ion mass spectrometry (ToF-SIMS) was used to distinguish between different proteins and their orientation by monitoring the changes in intensity of characteristic amino acid mass fragments. Amino acids distributed asymmetrically were used to calculate peak intensity ratios from ToF-SIMS data to determine the orientation of five different cysteine mutants of protein G B1 covalently attached to a maleimide surface. To study the effect of protein orientation on antibody binding, we formed multilayer protein films by binding IgG to protein G B1 films. Quartz crystal microbalance with dissipation monitoring (QCM-D) detected protein coverages of 69-130 ng/cm2 (theoretical mass of a monolayer of protein G B1 is 110-160 ng/cm2). QCM-D and X-ray photoelectron spectroscopy analysis revealed that packing density along with orientation affected the antibody binding process. Spectra from ToF-SIMS using large Ar gas cluster ion sources distinguished between different proteins in multilayer protein systems. A Monte Carlo algorithm was developed to predict protein orientation on surfaces. Two distinct orientations of protein G B1 adsorbed onto a hydrophobic surface were found and characterized as two mutually exclusive sets of amino acids on the outermost

  19. Surface modification of polyvinyl alcohol/malonic acid nanofibers by gaseous dielectric barrier discharge plasma for glucose oxidase immobilization

    Energy Technology Data Exchange (ETDEWEB)

    Afshari, Esmail, E-mail: e.afshari@mail.sbu.ac.ir [Laser and Plasma Research Institute, Shahid Beheshti University, Evin, 1983963113 Tehran (Iran, Islamic Republic of); Mazinani, Saeedeh [Amirkabir Nanotechnology Research Institute (ANTRI), Amirkabir University of Technology, 15875-4413, Tehran (Iran, Islamic Republic of); Ranaei-Siadat, Seyed-Omid [Protein Research Center, Shahid Beheshti University, Evin, 1983963113 Tehran (Iran, Islamic Republic of); Ghomi, Hamid [Laser and Plasma Research Institute, Shahid Beheshti University, Evin, 1983963113 Tehran (Iran, Islamic Republic of)

    2016-11-01

    Highlights: • We fabricated polyvinyl alcohol/malonic acid nanofibers using electrospinning. • The surface nanofibers were modified by gaseous (air, nitrogen, CO{sub 2} and argon) dielectric barrier discharge. • Among them, air plasma had the most significant effect on glucose oxidase immobilization. • Chemical analysis showed that after modification of nanofibers by air plasma, the carboxyl group increased. • After air plasma treatment, reusability and storage stability of glucose oxidase immobilized on nanofibers improved. - Abstract: Polymeric nanofiber prepares a suitable situation for enzyme immobilization for variety of applications. In this research, we have fabricated polyvinyl alcohol (PVA)/malonic acid nanofibers using electrospinning. After fabrication of nanofibers, the effect of air, nitrogen, CO{sub 2}, and argon DBD (dielectric barrier discharge) plasmas on PVA/malonic acid nanofibers were analysed. Among them, air plasma had the most significant effect on glucose oxidase (GOx) immobilization. Attenuated total reflectance-Fourier transform infrared (ATR-FTIR) spectrum analysis and X-ray photoelectron spectroscopy (XPS) results revealed that in case of air plasma modified nanofibers, the carboxyl groups on the surface are increased. The scanning electron microscopy (SEM) images showed that, after GOx immobilization, the modified nanofibers with plasma has retained its nanofiber structure. Finally, we analysed reusability and storage stability of GOx immobilized on plasma modified and unmodified nanofibers. The results were more satisfactory for modified nanofibers with respect to unmodified ones.

  20. Plasma immersion ion implantation of polyurethane shape memory polymer: Surface properties and protein immobilization

    Science.gov (United States)

    Cheng, Xinying; Kondyurin, Alexey; Bao, Shisan; Bilek, Marcela M. M.; Ye, Lin

    2017-09-01

    Polyurethane-type shape memory polymers (SMPU) are promising biomedical implant materials due to their ability to recover to a predetermined shape from a temporary shape induced by thermal activation close to human body temperature and their advantageous mechanical properties including large recovery strains and low recovery stresses. Plasma Immersion Ion Implantation (PIII) is a surface modification process using energetic ions that generates radicals in polymer surfaces leading to carbonisation and oxidation and the ability to covalently immobilise proteins without the need for wet chemistry. Here we show that PIII treatment of SMPU significantly enhances its bioactivity making SMPU suitable for applications in permanent implantable biomedical devices. Scanning Electron Microscopy (SEM), contact angle measurements, surface energy measurements, attenuated total reflection Fourier transform infrared (ATR-FTIR) spectroscopy and X-ray photoelectron spectroscopy (XPS) were used to characterise the PIII modified surface, including its after treatment aging kinetics and its capability to covalently immobilise protein directly from solution. The results show a substantial improvement in wettability and dramatic changes of surface chemical composition dependent on treatment duration, due to the generation of radicals and subsequent oxidation. The SMPU surface, PIII treated for 200s, achieved a saturated level of covalently immobilized protein indicating that a full monolayer coverage was achieved. We conclude that PIII is a promising and efficient surface modification method to enhance the biocompatibility of SMPU for use in medical applications that demand bioactivity for tissue integration and stability in vivo.

  1. Introduction of sulfate groups on poly(ethylene) surfaces by argon plasma immobilization of sodium alkyl sulfates

    NARCIS (Netherlands)

    Lens, J.P.; Lens, J.P.; Terlingen, J.G.A.; Terlingen, J.G.A.; Engbers, G.H.M.; Feijen, Jan

    1998-01-01

    Sulfate groups were introduced at the surface of poly(ethylene) (PE) samples. This was accomplished by immobilizing a precoated layer of either sodium 10-undecene sulfate (S11(:)) or sodium dodecane sulfate (SDS) on the polymeric surface by means of an argon plasma treatment. For this purpose,

  2. Use of activated carbon as a support medium for H{sub 2}S biofiltration and effect of bacterial immobilization on available pore surface

    Energy Technology Data Exchange (ETDEWEB)

    Ng, Y.L.; Yan, R.; Chen, X.G.; Geng, A.L.; Liang, D.T.; Koe, L.C.C. [Institute of Environmental Science and Engineering, Nanyang Technological Univ., Singapore (Singapore); Gould, W.D. [Environmental Lab., CANMET, Natural Resources Canada, Ottawa, ON (Canada)

    2004-12-01

    The use of support media for the immobilization of micro-organisms widely known to provide a surface for microbial growth and a shelter that protects the microorganisms from inhibitory compounds. In this study, activated carbon is used as a support medium for the immobilization of microorganisms enriched from municipal sewage activated sludge to remove gas-phase hydrogen sulfide (H{sub 2}S), a major odorous component of waste gas from sewage treatment plants. A series of designed experiments is used to examine the effect on bacteria-immobilized activated carbon (termed ''biocarbon'') due to physical adsorption, chemical reaction and microbial degradation in the overall removal of H{sub 2}S. H{sub 2}S breakthrough tests are conducted with various samples, including micro-immobilized carbon and Teflon discs, salts-medium-washed carbon, and ultra-pure water-washed carbon. The results show a higher removal capacity for the microbe-immobilized activated carbon compared with the activated carbon control in a batch biofilter column. The increase in removal capacity is attributed to the role played by the immobilized micro-organisms in metabolizing adsorbed sulfur and sulfur compounds on the biocarbon, hence releasing the adsorption sites for further H{sub 2}S uptake. The advantage for activated carbon serving as the support medium is to adsorb a high initial concentration of substrate and progressively release this for microbial degradation, hence acting as a buffer for the microorganisms. Results obtained from surface area and pore size distribution analyses of the biocarbon show a correlation between the available surface area and pore volume with the extent of microbial immobilization and H{sub 2}S uptake. The depletion of surface area and pore volume is seen as one of the factors which cause the onset of column breakthrough. Microbial growth retardation is due to the accumulation of metabolic products (i.e., sulfuric acid); and a lack of water and

  3. Immobilization of Phosphomolybdate Anions on the Surface of Magnetite Nanoparticles Modified with Quaternary Phosphonium Cations

    Directory of Open Access Journals (Sweden)

    M. Masteri-Farahani

    2013-06-01

    Full Text Available A new hybrid nanomaterial was developed by immobilization of phosphomolybdate anions on the surface of modified magnetite nanoparticles with quaternary phosphonium cations. Silica coated magnetic nanoparticles supported phosphonium cations, Ph3P+-SCMNPs, were prepared by covalent attachment of chloropropylsilyl groups on the surface of silica coated magnetite nanoparticles and next reaction with triphenylphosphine. Then, reaction of the prepared Ph3P+-SCMNPs nanomaterial with H3PMo12O40 resulted in the preparation of PMo-Ph3P+-SCMNPs hybrid nanomaterial. The PMo-Ph3P+-SCMNPs hybrid nanomaterial was characterized with different physicochemical methods such as FT-IR and ICP-AES spectroscopies, XRD, VSM, SEM, and TEM analyses. VSM analysis showed superparamagnetic properties of the prepared nanomaterial. TEM and SEM analyses indicated the aggregated nanoparticles with about 15 nm average size.

  4. Immobilization of simulated reducing agent at the surface of SiO2 fillers in dental composite resins.

    Science.gov (United States)

    Shibata, Satoki; Hirata, Isao; Nomura, Yuji; Shirai, Kenichi; Fujitani, Morioki; Shintani, Hideaki; Okazaki, Masayuki

    2007-07-01

    To reduce the leachability of reducing agents from composite resins, immobilization of a simulated reducing agent at the surface of SiO2 fillers was examined. SiO2 plates were immersed in 2% 3-aminopropyltriethoxy silane/ethanol solution, and then immersed in dimethyl sulfoxide with 0.25 wt% 4-dimethyl amino benzoic acid (DMABA), 2.0 wt% 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride, and 0.5 wt% N-hydroxysuccinimide. Wide-scan spectrum of X-ray photoelectron spectroscopy did not detect carbon contamination. However, narrow scan detected an O=C-N peak at 399.8 eV, suggesting that DMABA could be immobilized on silane-coupled SiO2 plates. Further, surface plasmon resonance analysis indicated the adsorption of MMA at the surface of reducing agent-immobilized plate.

  5. Surface Redox Chemistry of Immobilized Nanodiamond: Effects of Particle Size and Electrochemical Environment

    Science.gov (United States)

    Gupta, S.; McDonald, B.; Carrizosa, S. B.

    2017-07-01

    The size of the diamond particle is tailored to nanoscale (nanodiamond, ND), and the ND surface is engineered targeting specific (electrochemical and biological) applications. In this work, we investigated the complex surface redox chemistry of immobilized ND layer on conductive boron-doped diamond electrode with a broad experimental parameter space such as particle size (nano versus micron), scan rate, pH (cationic/acidic versus anionic/basic), electrolyte KCl concentration (four orders of magnitude), and redox agents (neutral and ionic). We reported on the significant enhancement of ionic currents while recording reversible oxidation of neutral ferrocene methanol (FcMeOH) by almost one order of magnitude than traditional potassium ferricyanide (K3Fe(CN)6) redox agent. The current enhancement is inversely related to ND particle diameter in the following order: 1 μm << 1000 nm < 100 nm < 10 nm ≤ 5 nm < 2 nm. We attribute the current enhancement to concurrent electrocatalytic processes, i.e. the electron transfer between redox probes and electroactive surface functional (e.g. hydroxyl, carboxyl, epoxy) moieties and the electron transfer mediated by adsorbed FcMeOH+ (or Fe(CN) 6 3+ ) ions onto ND surface. The first process is pH dependent since it depends upon ND surface functionalities for which the electron transfer is coupled to proton transfer. The adsorption mediated process is observed most apparently at slower scan rates owing to self-exchange between adsorbed FcMeOH+ ions and FcMeOH redox agent molecules in diffusion-limited bulk electrolyte solution. Alternatively, it is hypothesized that the surface functionality and defect sites ( sp 2-bonded C shell and unsaturated bonds) give rise to surface electronic states with energies within the band gap (midgap states) in undoped ND. These surface states serve as electron donors (and acceptors) depending upon their bonding (and antibonding) character and, therefore, they can support electrocatalytic redox

  6. Surface modification of polypropylene nonwoven fabrics via covalent immobilization of nonionic sugar-based surfactants

    Energy Technology Data Exchange (ETDEWEB)

    Xin, Zhirong, E-mail: xinzhirong2012@126.com; Yan, Shunjie; Ding, Jiaotong; Yang, Zongfeng; Du, Binbin; Du, Shanshan

    2014-05-01

    Graphical abstract: - Highlights: • Amphiphilic N-alkyl-1-amino-1-deoxy-D-glucitol were prepared. • The pGMA-grafted membranes were obtained via photo-grafting of glycidyl methacrylate. • Amphiphilic C{sub n}AG were covalent immobilized onto the pGMA-grafted membranes. • The C{sub n}AG-grafted membranes obviously suppressed protein adsorption and platelet adhesion. - Abstract: Amphiphilic N-alkyl-1-amino-1-deoxy-D-glucitol (C{sub n}AG, n = 8, 12) were successfully prepared. Polypropylene nonwoven fabrics (PP{sub NWF}) were grafted with glycidyl methacrylate (GMA) via a technique of UV-induced graft polymerization combined with plasma pre-treatment, and then PP{sub NWF}-g-GMA was used for the covalent immobilization of C{sub n}AG. The surface graft polymerization was confirmed by ATR-FTIR and XPS, respectively. Effect of grafting parameters, e.g., acetone content, monomer concentration and UV irradiation time on the grafting density of GMA was investigated. And the hemocompatibility of the modified PP{sub NWF} was evaluated by protein adsorption and platelet adhesion. It was founded that the C{sub n}AG-modified substrates greatly suppressed protein adsorption and platelet adhesion compared with the native and pGMA-grafted PP{sub NWF}.

  7. Nano- and femtosecond UV laser pulses to immobilize biomolecules onto surfaces with preferential orientation

    Science.gov (United States)

    Lettieri, S.; Avitabile, A.; Della Ventura, B.; Funari, R.; Ambrosio, A.; Maddalena, P.; Valadan, M.; Velotta, R.; Altucci, C.

    2014-10-01

    By relying on the photonic immobilization technique of antibodies onto surfaces, we realized portable biosensors for light molecules based on the use of quartz crystal microbalances, given the linear dependence of the method on the laser pulse intensity. Here, we compare the quality of the anchoring method when using nanosecond (260 nm, 25 mJ/pulse, 5 ns, 10 Hz rep. rate) and femtosecond (258 nm, 25 μJ/pulse, 150 fs, 10 kHz rep. rate) laser source, delivering the same energy to the sample with the same average power. As a reference, we also tethered untreated antibodies by means of the passive adsorption. The results are striking: When the antibodies are irradiated with the femtosecond pulses, the deposition on the gold plate is much more ordered than in the other two cases. The effects of UV pulses irradiation onto the antibodies are also analyzed by measuring absorption and fluorescence and suggest the occurrence of remarkable degradation when nanosecond pulses are used likely induced by a larger thermal coupling. In view of the high average power required to activate the antibodies for the achievement of the photonic immobilization technique, we conclude that femtosecond rather than nanosecond laser pulses have to be used.

  8. Surface modification of polypropylene nonwoven fabrics via covalent immobilization of nonionic sugar-based surfactants

    International Nuclear Information System (INIS)

    Xin, Zhirong; Yan, Shunjie; Ding, Jiaotong; Yang, Zongfeng; Du, Binbin; Du, Shanshan

    2014-01-01

    Graphical abstract: - Highlights: • Amphiphilic N-alkyl-1-amino-1-deoxy-D-glucitol were prepared. • The pGMA-grafted membranes were obtained via photo-grafting of glycidyl methacrylate. • Amphiphilic C n AG were covalent immobilized onto the pGMA-grafted membranes. • The C n AG-grafted membranes obviously suppressed protein adsorption and platelet adhesion. - Abstract: Amphiphilic N-alkyl-1-amino-1-deoxy-D-glucitol (C n AG, n = 8, 12) were successfully prepared. Polypropylene nonwoven fabrics (PP NWF ) were grafted with glycidyl methacrylate (GMA) via a technique of UV-induced graft polymerization combined with plasma pre-treatment, and then PP NWF -g-GMA was used for the covalent immobilization of C n AG. The surface graft polymerization was confirmed by ATR-FTIR and XPS, respectively. Effect of grafting parameters, e.g., acetone content, monomer concentration and UV irradiation time on the grafting density of GMA was investigated. And the hemocompatibility of the modified PP NWF was evaluated by protein adsorption and platelet adhesion. It was founded that the C n AG-modified substrates greatly suppressed protein adsorption and platelet adhesion compared with the native and pGMA-grafted PP NWF

  9. Covalent and density-controlled surface immobilization of E-cadherin for adhesion force spectroscopy.

    Directory of Open Access Journals (Sweden)

    Dagmar Fichtner

    Full Text Available E-cadherin is a key cell-cell adhesion molecule but the impact of receptor density and the precise contribution of individual cadherin ectodomains in promoting cell adhesion are only incompletely understood. Investigating these mechanisms would benefit from artificial adhesion substrates carrying different cadherin ectodomains at defined surface density. We therefore developed a quantitative E-cadherin surface immobilization protocol based on the SNAP-tag technique. Extracellular (EC fragments of E-cadherin fused to the SNAP-tag were covalently bound to self-assembled monolayers (SAM of thiols carrying benzylguanine (BG head groups. The adhesive functionality of the different E-cadherin surfaces was then assessed using cell spreading assays and single-cell (SCSF and single-molecule (SMSF force spectroscopy. We demonstrate that an E-cadherin construct containing only the first and second outmost EC domain (E1-2 is not sufficient for mediating cell adhesion and yields only low single cadherin-cadherin adhesion forces. In contrast, a construct containing all five EC domains (E1-5 efficiently promotes cell spreading and generates strong single cadherin and cell adhesion forces. By varying the concentration of BG head groups within the SAM we determined a lateral distance of 5-11 nm for optimal E-cadherin functionality. Integrating the results from SCMS and SMSF experiments furthermore demonstrated that the dissolution of E-cadherin adhesion contacts involves a sequential unbinding of individual cadherin receptors rather than the sudden rupture of larger cadherin receptor clusters. Our method of covalent, oriented and density-controlled E-cadherin immobilization thus provides a novel and versatile platform to study molecular mechanisms underlying cadherin-mediated cell adhesion under defined experimental conditions.

  10. Antifungal coatings by caspofungin immobilization onto biomaterials surfaces via a plasma polymer interlayer.

    Science.gov (United States)

    Griesser, Stefani S; Jasieniak, Marek; Coad, Bryan R; Griesser, Hans J

    2015-12-14

    Not only bacteria but also fungal pathogens, particularly Candida species, can lead to biofilm infections on biomedical devices. By covalent grafting of the antifungal drug caspofungin, which targets the fungal cell wall, onto solid biomaterials, a surface layer can be created that might be able to provide long-term protection against fungal biofilm formation. Plasma polymerization of propionaldehyde (propanal) was used to deposit a thin (∼20 nm) interfacial bonding layer bearing aldehyde surface groups that can react with amine groups of caspofungin to form covalent interfacial bonds for immobilization. Surface analyses by x-ray photoelectron spectroscopy and time-of-flight secondary ion mass spectrometry confirmed the intended grafting and uniformity of the coatings, and durability upon extended washing. Testing for fungal cell attachment and ensuing biofilm formation showed that caspofungin retained activity when covalently bound onto surfaces, disrupting colonizing Candida cells. Mammalian cytotoxicity studies using human primary fibroblasts indicated that the caspofungin-grafted surfaces were selective in eliminating fungal cells while allowing attachment and spreading of mammalian cells. These in vitro data suggest promise for use as antifungal coatings, for example, on catheters, and the use of a plasma polymer interlayer enables facile transfer of the coating method onto a wide variety of biomaterials and biomedical devices.

  11. Screening hydroxyapatite for cadmium and lead immobilization in aqueous solution and contaminated soil: The role of surface area.

    Science.gov (United States)

    Li, Hongying; Guo, Xisheng; Ye, Xinxin

    2017-02-01

    Hydroxyapatite (HAP) has been widely used to immobilize many cationic metals in water and soils. The specific reason why an increase in the surface area of HAP enhances cadmium (Cd) uptake, but has no effect on lead (Pb) uptake, is not clear. The aim of this study was to determine the factors causing the differences in sorption behavior between Cd and Pb by evaluating HAPs with different surface areas. We synthesized HAPs with two different surface areas, which were characterized by X-ray diffraction, N 2 adsorption, and scanning electron microscopy, and then evaluated them as sorbents for Cd and Pb removal by testing in single and binary systems. The sorption capacity of large surface area HAP (1.85mmol/g) for Cd in the single-metal system was higher than that of small surface area HAP (0.64mmol/g), but there were no differences between single- and binary-metal solutions containing Pb. After the Cd experiments, the HAP retained a stable structure and intact morphology, which promotes the accessibility of reactive sites for Cd. However, a newly formed precipitate covered the surface and blocked the channels in the presence of Pb, which reduced the number of potential adsorption sites on HAP for Cd and Pb. Remediation experiments using Cd- and Pb-contaminated soil produced similar results to the solution tests. These results indicate that alterations of the structure and morphology during the reaction is an important factor influencing metal sorption to HAP. Copyright © 2016. Published by Elsevier B.V.

  12. Decolorization of textile dye RB19 using volcanic rock matrix immobilized Bacillus thuringiensis cells with surface displayed laccase.

    Science.gov (United States)

    Wan, Juan; Sun, Xiaowen; Liu, Cheng; Tang, Mengjun; Li, Lin; Ni, Hong

    2017-06-01

    A triplicate volcanic rock matrix-Bacillus thuringiensis-laccase WlacD (VRMs-Bt-WlacD) dye decolorization system was developed. WlacD was displayed on the B. thuringiensis MB174 cell surface to prepare a whole-cell laccase biocatalyst by using two repeat N-terminal domains of autolysin Mbg (Mbgn) 2 as the anchoring motif. Immunofluorescence microscopic assays confirmed that the fusion protein (Mbgn) 2 -WlacD was anchored on the surface of the recombinant B. thuringiensis MB174. After optimization by a single factor test, L 9 (3 4 )-orthogonal test, Plackett-Burman test, steepest ascent method, and Box-Behnken response surface methodology, the whole-cell specific laccase activity of B. thuringiensis MB174 was improved to 555.2 U L -1 , which was 2.25 times than that of the primary culture condition. Optimized B. thuringiensis MB174 cells were further adsorbed by VRMs to prepare VRMs-Bt-WlacD, an immobilized whole-cell laccase biocatalyst. Decolorization capacity of as-prepared VRMs-Bt-WlacD toward an initial concentration of 500 mg L -1 of an textile dye reactive blue 19 (RB19) aqueous solution reached 72.36% at a solid-to-liquid ratio of 10 g-100 mL. Repeated decolorization-activation operations showed the high decolorization capacity of VRMs-Bt-WlacD and have the potential for large-scale or continuous operations.

  13. Decolorization of industrial synthetic dyes using engineered Pseudomonas putida cells with surface-immobilized bacterial laccase

    Science.gov (United States)

    2012-01-01

    Background Microbial laccases are highly useful in textile effluent dye biodegradation. However, the bioavailability of cellularly expressed or purified laccases in continuous operations is usually limited by mass transfer impediment or enzyme regeneration difficulty. Therefore, this study develops a regenerable bacterial surface-displaying system for industrial synthetic dye decolorization, and evaluates its effects on independent and continuous operations. Results A bacterial laccase (WlacD) was engineered onto the cell surface of the solvent-tolerant bacterium Pseudomonas putida to construct a whole-cell biocatalyst. Ice nucleation protein (InaQ) anchor was employed, and the ability of 1 to 3 tandemly aligned N-terminal repeats to direct WlacD display were compared. Immobilized WlacD was determined to be surface-displayed in functional form using Western blot analysis, immunofluorescence microscopy, flow cytometry, and whole-cell enzymatic activity assay. Engineered P. putida cells were then applied to decolorize the anthraquinone dye Acid Green (AG) 25 and diazo-dye Acid Red (AR) 18. The results showed that decolorization of both dyes is Cu2+- and mediator-independent, with an optimum temperature of 35°C and pH of 3.0, and can be stably performed across a temperature range of 15°C to 45°C. A high activity toward AG25 (1 g/l) with relative decolorization values of 91.2% (3 h) and 97.1% (18 h), as well as high activity to AR18 (1 g/l) by 80.5% (3 h) and 89.0% (18 h), was recorded. The engineered system exhibited a comparably high activity compared with those of separate dyes in a continuous three-round shake-flask decolorization of AG25/AR18 mixed dye (each 1 g/l). No significant decline in decolorization efficacy was noted during first two-rounds but reaction equilibriums were elongated, and the residual laccase activity eventually decreased to low levels. However, the decolorizing capacity of the system was easily retrieved via a subsequent 4-h

  14. Decolorization of industrial synthetic dyes using engineered Pseudomonas putida cells with surface-immobilized bacterial laccase

    Directory of Open Access Journals (Sweden)

    Wang Wei

    2012-06-01

    Full Text Available Abstract Background Microbial laccases are highly useful in textile effluent dye biodegradation. However, the bioavailability of cellularly expressed or purified laccases in continuous operations is usually limited by mass transfer impediment or enzyme regeneration difficulty. Therefore, this study develops a regenerable bacterial surface-displaying system for industrial synthetic dye decolorization, and evaluates its effects on independent and continuous operations. Results A bacterial laccase (WlacD was engineered onto the cell surface of the solvent-tolerant bacterium Pseudomonas putida to construct a whole-cell biocatalyst. Ice nucleation protein (InaQ anchor was employed, and the ability of 1 to 3 tandemly aligned N-terminal repeats to direct WlacD display were compared. Immobilized WlacD was determined to be surface-displayed in functional form using Western blot analysis, immunofluorescence microscopy, flow cytometry, and whole-cell enzymatic activity assay. Engineered P. putida cells were then applied to decolorize the anthraquinone dye Acid Green (AG 25 and diazo-dye Acid Red (AR 18. The results showed that decolorization of both dyes is Cu2+- and mediator-independent, with an optimum temperature of 35°C and pH of 3.0, and can be stably performed across a temperature range of 15°C to 45°C. A high activity toward AG25 (1 g/l with relative decolorization values of 91.2% (3 h and 97.1% (18 h, as well as high activity to AR18 (1 g/l by 80.5% (3 h and 89.0% (18 h, was recorded. The engineered system exhibited a comparably high activity compared with those of separate dyes in a continuous three-round shake-flask decolorization of AG25/AR18 mixed dye (each 1 g/l. No significant decline in decolorization efficacy was noted during first two-rounds but reaction equilibriums were elongated, and the residual laccase activity eventually decreased to low levels. However, the decolorizing capacity of the system was easily retrieved

  15. Spectral surface plasmon resonance imaging for the detection of clenbuterol via three-dimensional immobilization of bioprobes.

    Science.gov (United States)

    Yao, Manwen; Wu, Yichuan; Fang, Xiangyi; Yang, Yucong; Liu, Hongjun

    2015-04-15

    A method of immobilizing clenbuterol (CLEN) on the sensor chip for spectral surface plasmon resonance imaging (SPRi) was experimentally investigated. The bioprobes on the sensor chip were prepared by immobilizing bovine serum albumin (BSA) protein and conjugating CLEN molecules to BSA, which provides more active points and free orientations for specific binding. The calibration curve showed that the wavelength resonance shift decreased as the concentration of CLEN analyte increased, consistent with the inhibition principle. The limit of detection (LOD) was estimated to be 6.32 μg/ml. This method proved to be highly specific, high throughput, label free, and operationally convenient. Copyright © 2015 Elsevier Inc. All rights reserved.

  16. Three Dimensional Immobilization of Beta-Galactosidase on a Silicon Surface (Preprint)

    National Research Council Canada - National Science Library

    Luckarift, Heather R; Betancor, Lorena; Spain, Jim C

    2006-01-01

    .... The entrapment of enzymes within silica-based nanospheres formed through silicification reactions provides high loading capacities for enzyme immobilization, resulting in high volumetric activity...

  17. Protein immobilization on the surface of liposomes via carbodiimide activation in the presence of N-hydroxysulfosuccinimide.

    Science.gov (United States)

    Bogdanov, A A; Klibanov, A L; Torchilin, V P

    1988-04-25

    A method of the covalent immobilization of proteins on the surface of liposomes, containing 10% (by mol) of N-glutaryl phosphatidylethanolamine, is described. Carboxylic groups of liposomal N-glutaryl phosphatidylethanolamine were activated in the presence of water-soluble carbodiimide and N-hydroxysulfosuccinimide and reacted subsequently with protein amino groups. The liposome-protein conjugates formed contained up to 5 x 10(-4) mol protein/mol lipid. Lectins (RCA1 and WGA) upon immobilization on liposomes retained saccharide specificity and the ability to agglutinate red blood cells. The immobilization of mouse monoclonal IgG in a ratio of 3.5 x 10(-4) mol IgG/mol lipid was achieved. The liposome activation in the absence of N-hydroxysulfosuccinimide resulted in a 2-fold decrease of protein coupling yields.

  18. Protein-Nanoparticle Interactions: Improving Immobilized Lytic Enzyme Activity and Surface Energy Effects

    Science.gov (United States)

    Downs, Emily Elizabeth

    -particle interactions to protein-protein interactions and were thicker with greater surface energy, which resulted in the recovery of secondary structure in the outermost layer. To help understand the impact of protein structure on nano-bio conjugate interactions, a listeria specific protein was used. This system was chosen as it has applications in the food industry in preventing bacterial contamination. The insertion of an amino acid linker between the enzymatic and binding domain of the protein improved the flexibility between domains, leading to increased adsorption, and improved activity in both cell-wall and plating assays. Additionally, linker modified protein incorporated into the silica-polymer nanocomposite showed significant activity in a real-world example of contaminated lettuce. This thesis study has isolated the impact of surface energy and protein flexibility on protein adsorption and structure. Particle surface energy affects adsorbed protein concentration and conformation. Coupled with protein surface charge, surface energy was also found to dictate multilayer thickness. The conformational flexibility of the protein was shown to help in controlling not only protein adsorption concentration but also in retaining protein activity after immobilization. Also, a controllable synthesis method for particles with adjustable surface energy, an ideal platform for studying protein-particle interactions, has been established.

  19. Evaluation of the effectiveness factor along immobilized enzyme fixed-bed reactors: design of a reactor with naringinase covalently immobilized into glycophase-coated porous glass

    Energy Technology Data Exchange (ETDEWEB)

    Manjon, A.; Iborra, J.L.; Gomez, J.L.; Gomez, E.; Bastida, J.; Bodalo, A.

    1987-09-01

    A design equation is presented for packed-bed reactors containing immobilized enzymes in spherical porous particles with internal diffusion effects and obeying reversible one-intermediate Michaelis-Menten kinetics. The equation is also able to explain irreversible and competitive product inhibition kinetics. It allows the axial substrate profiles to be calculated and the dependence of the effectiveness factor along the reactor length to be continuously evaluated. The design equation was applied to explain the behavior of naringinase immobilized in Glycophase-coated porous glass operating in a packed-bed reactor and hydrolyzing both p-nitrophenyl-alpha-L-rhamnoside and naringin. The theoretically predicted results were found to fit well with experimentally measured values. (Refs. 28).

  20. Horseradish peroxidase immobilized on copper surfaces and applications in selective electrocatalysis of p-dihydroxybenzene

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Chuntao, E-mail: tsyj1992@126.com [Department of Chemistry, Taiyuan Normal University, Taiyuan 030031 (China); Institute of Energy and Environmental Electrochemistry, Taiyuan Normal University, Taiyuan 030031 (China); Luo, Xiaoxiao [Department of Natural Science, Michigan State University, MI 48823,USA (United States); Jia, Zehui [Department of Chemistry, Taiyuan Normal University, Taiyuan 030031 (China); Institute of Energy and Environmental Electrochemistry, Taiyuan Normal University, Taiyuan 030031 (China); Shi, Qinghua; Zhu, Ritao [Department of Chemistry, Taiyuan Normal University, Taiyuan 030031 (China)

    2017-06-01

    Abstract: Horseradish Peroxidase (HRP) was immobilized on copper surfaces with the linker of L-Cysteine (L-Cys) self-assembled films to form Cu/L-Cys/HRP electrodes. The activity of HRP can be preserved by the Cu/L-Cys self-assembled films. The Cu/L-Cys/HRP electrodes can be used for the selective electrocatalytic oxidase of p-dihydroxybenzen in absent of H{sub 2}O{sub 2}. The optimum pH for electrocatalyzing p-dihydroxybenzen was 5.5 or 7.0, which corresponds to the isoelectric points of L-Cys and HRP, respectively. X-ray photoelectron spectroscopy (XPS) provided the evidence that L-Cys linked with Cu surface by the Cu− S bond. Fourier transform infrared spectroscopy (FTIR) analyses indicated that aromatic plane of p-dihydroxybenzen was connected parallel to porphyrin ring of heme in HRP. Quantum chemical calculation of density functional theory (DFT) revealed that symmetry of molecular structure and minimum space steric hindrance for p-dihydroxybenzen were benefit to combination with HRP. Moreover, the lowest energy of LUMO and most negative charges of oxygen atom on hydroxyl group of p-dihydroxybenzen were advantage to lose the hydrogen atom of hydroxyl group to be oxided.

  1. Immobilization of oligonucleotide probes on silicon surfaces using biotin–streptavidin system examined with microscopic and spectroscopic techniques

    Energy Technology Data Exchange (ETDEWEB)

    Awsiuk, K., E-mail: kamil.awsiuk@uj.edu.pl [M. Smoluchowski Institute of Physics, Jagiellonian University, Reymonta 4, Kraków 30-059 (Poland); Rysz, J. [M. Smoluchowski Institute of Physics, Jagiellonian University, Reymonta 4, Kraków 30-059 (Poland); Petrou, P. [Institute of Nuclear and Radiological Sciences and Technology, Energy and Safety, NCSR “Demokritos”, End Patriarchou Gregoriou Str., Aghia Paraskevi 15310 (Greece); Budkowski, A. [M. Smoluchowski Institute of Physics, Jagiellonian University, Reymonta 4, Kraków 30-059 (Poland); Bernasik, A. [Faculty of Physics and Applied Computer Science, AGH-University of Science and Technology, Al. Mickiewicza 30, Kraków 30-059 (Poland); Kakabakos, S. [Institute of Nuclear and Radiological Sciences and Technology, Energy and Safety, NCSR “Demokritos”, End Patriarchou Gregoriou Str., Aghia Paraskevi 15310 (Greece); Marzec, M.M. [Faculty of Physics and Applied Computer Science, AGH-University of Science and Technology, Al. Mickiewicza 30, Kraków 30-059 (Poland); Raptis, I. [Institute for Advanced Materials, Physicochemical Processes, Nanotechnology and Microsystems, NCSR “Demokritos”, End Patriarchou Gregoriou Str., Aghia Paraskevi 15310 (Greece)

    2014-01-30

    To immobilize effectively oligonucleotide probes on SiO{sub 2} modified with (3-aminopropyl)triethoxysilane, four procedures based on streptavidin–biotin system are compared with Atomic Force Microscopy, Angle-Resolved X-ray Photoelectron Spectroscopy and Time-of-Flight Secondary Ion Mass Spectrometry. The first approach involves: adsorption of biotinylated Bovine Serum Albumin, blocking free surface sites with BSA, binding of streptavidin and biotinylated oligonucleotide (b-oligo). Final steps are exchanged in the second procedure with immobilization of preformed streptavidin–b-oligo conjugate. The third approach consists of streptavidin adsorption, blocking with BSA and b-oligo binding. Finally, streptavidin–b-oligo conjugate is immobilized directly within the fourth method. Surface coverage with biomolecules, determined from ARXPS, accords with average AFM height, and is anti-correlated with the intensity of Si+ ions. Higher biomolecular coverage was achieved during the last steps of the first (2.45(±0.38) mg/m{sup 2}) and second (1.31(±0.22) mg/m{sup 2}) approach, as compared to lower surface density resulting from the third (0.58(±0.20) mg/m{sup 2}) and fourth (0.41(±0.11) mg/m{sup 2}) method. Phosphorus atomic concentration indicates effectiveness of oligonucleotide immobilization. Secondary ions intensities, characteristic for oligonucleotides, streptavidin, BSA, and proteins, allow additional insight into overlayer composition. These measurements verify the ARXPS results and show the superiority of the first two immobilization approaches in terms of streptavidin and oligonucleotide density achieved onto the surface.

  2. Magnetic resonance imaging for radiotherapy planning of brain cancer patients using immobilization and surface coils

    Science.gov (United States)

    Hanvey, S.; Glegg, M.; Foster, J.

    2009-09-01

    This study investigated the compatibility of a head and neck immobilization device with magnetic resonance imaging (MRI). The immobilization device is used to position a patient in the same way as when receiving a computed tomography (CT) scan for radiotherapy planning and radiation treatment. The advantage of using immobilization in MR is improved accuracy in CT/MR image registration enabling greater confidence in the delineation of structures. The main practical difficulty in using an immobilization device in MRI is that physical constraints make their use incompatible with head imaging coils. Within this paper we describe a method for MR imaging of the brain which allows the use of head and neck immobilization devices. By a series of image quality tests we obtained the same or better image quality as a multi-channel head coil.

  3. Immobilization of BSA on ionic liquid functionalized magnetic Fe3O4nanoparticles for use in surface imprinting strategy.

    Science.gov (United States)

    Qian, Liwei; Sun, Jiexuan; Hou, Chen; Yang, Jinfan; Li, Yongwei; Lei, Dan; Yang, Miaoxiu; Zhang, Sufeng

    2017-06-01

    Combining template immobilization with surface imprinting technology is an effective strategy to overcome the difficulties associated with macromolecular template removal and to achieve high specific recognition ability. In this work, ionic liquid functionalized Fe 3 O 4 nanoparticles were prepared via a simple two-step modification process and were used as substrate to immobilize bovine serum albumin (BSA). The zeta potential study revealed immobilization of BSA on the nanoparticles through multiple interactions, and the immobilization capacity was about nine times higher compared with that of bare Fe 3 O 4 . Subsequently, dopamine was utilized as functional monomer to prepare BSA surface imprinted nanoparticles. Fourier transform infrared spectroscopy, thermo-gravimetric analysis and transmission electron microscopy verified the successful preparation of BSA imprinted nanoparticles with core-shell structure. The influence of imprinted layer thickness on recognition ability of imprinted nanoparticles was investigated, and the results suggested that 20nm was an optimum thickness to achieve the best recognition ability. The adsorption isotherm studies showed that the imprinted nanoparticles had a significantly higher adsorption capacity and stronger binding affinity than the non-imprinted ones. Furthermore, the selective as well as the competitive adsorption studies revealed higher selectivity and recognition ability of the imprinted nanoparticles for BSA. Therefore, the proposed strategy is an effective way to obtain protein imprinted polymers with high adsorption capacity and good recognition ability, thus would be beneficial for the further development and application of protein imprinting technology. Copyright © 2017 Elsevier B.V. All rights reserved.

  4. Immobilization of chitosan gel with cross-linking reagent on PNIPAAm gel/PP nonwoven composites surface

    International Nuclear Information System (INIS)

    Chen, K.-S.; Ku, Y.-A.; Lee, C.-H.; Lin, H.-R.; Lin, F.-H.; Chen, T.-M.

    2005-01-01

    This study was to immobilize chitosan (CS), which is a biodegradable and antibacterial polymer, on poly(N-isopropylacrylamide) (PNIPAAm) gel/polypropylene (PP) nonwoven composites surface for wound dressing applications. PP nonwoven has been extensively used due to its porosity, allowing ventilation, high surface area and excellent mechanical properties. However, the hydrophobic surface of PP nonwoven limits its applications; in this study, we used the plasma-activation treatment and subsequently UV-light graft polymerization of NIPAAm gel to improve its hydrophilicity. Chitosan was immobilized onto PNIPAAm gel/PP nonwoven composites surface using the cross-linking agent, glutaraldehyde (GA). This complex was characterized by scanning electron microscopy (SEM). The results indicated that the wettability of the composite was improved after plasma treatment and photo-induced graft polymerization and chitosan was successfully immobilized onto the surface of PNIPAAm gel/PP nonwoven composites through cross-linking process. Finally, the preliminary result shows that chitosan hydrogels displayed antibacterial ability to Escherichia coli and Staphylococcus aureus. The (3-(4, 5-dimethylthiazolyl-2)-2, 5-diphenyltetrazolium bromide) (MTT) method indicated that the porous chitosan sponge exhibited good biocompatibility to fibroblast cells

  5. Study on interactions of human IgG with immobilized anti-IgG or recombinant Staphylococcal protein A using surface plasmon resonance spectrometry

    Directory of Open Access Journals (Sweden)

    Bakhmachuk A. O.

    2016-02-01

    Full Text Available Aim. Comparison of the IgG-binding activity of recombinant Staphylococcal protein A with introduced C-terminal cysteine residue (SPA-Cys or goat anti-human IgG antibodies (anti-IgG after their immobilization on a gold sensor surface of surface plasmon resonance (SPR spectrometer. Methods. SPA-Cys or anti-IgG were immobilized on a gold sensor surface to form two variants of a bioselective element of the immunosensor. SPR spectrometry was used for the detection of IgG-binding activity of the immobilized proteins. Results.The SPR sensor response to the immobilization of anti-IgG was more than two times higher than that at the immobilization of SPA-Cys. However, there is almost the double advantage for SPA-Cys in the number of immobilized molecules. Moreover, the bioselective element of the immunosensor based on SPA-Cys showed a much better capability of binding Ig than bioselective element based on anti-IgG. Conclusions.The study on the immobilization of SPA-Cys or anti-IgG on the sensor surface of SPR spectrometer, and the interactions of immobilized proteins with human IgG demonstrated obvious advantages of SPA-Cys.

  6. Mechanism of the immobilization of surfactants on polymeric surfaces by means of an argon plasma treatment: influence of the chemical structure of surfactant substrate

    NARCIS (Netherlands)

    Lens, J.P.; Lens, J.P.; Terlingen, J.G.A.; Terlingen, J.G.A.; Engbers, G.H.M.; Feijen, Jan

    1998-01-01

    In this article, a study on the mechanism of the immobilization of surfactants on polymeric surfaces by means of an argon plasma treatment is described. The unsaturated surfactant sodium 10-undecenoate [C11(:)] and the saturated surfactant sodium dodecanoate (C12) were immobilized on poly(ethylene)

  7. Interfacial biocatalysis on charged and immobilized substrates: the roles of enzyme and substrate surface charge.

    Science.gov (United States)

    Feller, Bob E; Kellis, James T; Cascão-Pereira, Luis G; Robertson, Channing R; Frank, Curtis W

    2011-01-04

    An enzyme charge ladder was used to examine the role of electrostatic interactions involved in biocatalysis at the solid-liquid interface. The reactive substrate consisted of an immobilized bovine serum albumin (BSA) multilayer prepared using a layer-by-layer technique. The zeta potential of the BSA substrate and each enzyme variant was measured to determine the absolute charge in solution. Enzyme adsorption and the rate of substrate surface hydrolysis were monitored for the enzyme charge ladder series to provide information regarding the strength of the enzyme-substrate interaction and the rate of interfacial biocatalysis. First, each variant of the charge ladder was examined at pH 8 for various solution ionic strengths. We found that for positively charged variants the adsorption increased with the magnitude of the charge until the surface became saturated. For higher ionic strength solutions, a greater positive enzyme charge was required to induce adsorption. Interestingly, the maximum catalytic rate was not achieved at enzyme saturation but at an invariable intermediate level of adsorption for each ionic strength value. Furthermore, the maximum achievable reaction rate for the charge ladder was larger for higher ionic strength values. We propose that diffusion plays an important role in interfacial biocatalysis, and for strong enzyme-substrate interaction, the rate of diffusion is reduced, leading to a decrease in the overall reaction rate. We investigated the effect of substrate charge by varying the solution pH from 6.1 to 8.7 and by examining multiple ionic strength values for each pH. The same intermediate level of adsorption was found to maximize the overall reaction rate. However, the ionic strength response of the maximum achievable rate was clearly dependent on the pH of the experiment. We propose that this observation is not a direct effect of pH but is caused by the change in substrate surface charge induced by changing the pH. To prove this

  8. Reducing Escherichia coli growth on a composite biomaterial by a surface immobilized antimicrobial peptide

    Energy Technology Data Exchange (ETDEWEB)

    Buckholtz, Gavin A.; Reger, Nina A. [Department of Chemistry and Biochemistry, Duquesne University, Pittsburgh, PA 15282 (United States); Anderton, William D.; Schimoler, Patrick J. [Orthopaedic Biomechanics Research Laboratory, Allegheny General Hospital, Pittsburgh, PA 15212 (United States); Roudebush, Shana L.; Meng, Wilson S. [Division of Pharmaceutical Sciences, Duquesne University, Pittsburgh, PA 15282 (United States); Miller, Mark C. [Orthopaedic Biomechanics Research Laboratory, Allegheny General Hospital, Pittsburgh, PA 15212 (United States); Gawalt, Ellen S., E-mail: gawalte@duq.edu [Department of Chemistry and Biochemistry, Duquesne University, Pittsburgh, PA 15282 (United States); McGowan Institute for Regenerative Medicine, University of Pittsburgh, Pittsburgh, PA 15219 (United States)

    2016-08-01

    A new composite bioceramic consisting of calcium aluminum oxide (CaAlO) and hydroxyapatite (HA) was functionalized with the synthetic antimicrobial peptide Inverso-CysHHC10. CaAlO is a bioceramic that can be mold cast easily and quickly at room temperature. Improved functionality was previously achieved through surface reactions. Here, composites containing 0–5% HA (by mass) were prepared and the elastic modulus and modulus of rupture were mechanically similar to non-load bearing bone. The addition of hydroxyapatite resulted in increased osteoblast attachment (> 180%) and proliferation (> 140%) on all composites compared to 100% CaAlO. Antimicrobial peptide (AMP) immobilization was achieved using an interfacial alkene-thiol click reaction. The linked AMP persisted on the composite (> 99.6% after 24 h) and retained its activity against Escherichia coli based on N-phenylnaphthylamine uptake and bacterial turbidity tests. Overall, this simple scaffold system improves osteoblast activity and reduces bacterial activity. - Highlights: • Calcium aluminum oxide and hydroxyapatite were cast into a composite material. • Osteoblast attachment and proliferation were significantly increased on composites. • An active antimicrobial peptide was linked to and remained stable on the composite. • Bacterial turbidity and NPN uptake tests showed modified composites had an effect equal to a 10 μM Inverso-CysHHC10 solution. • Antimicrobial peptide linkage did not affect the increased osteoblast performance.

  9. Interaction force measurement between E. coli cells and nanoparticles immobilized surfaces by using AFM.

    Science.gov (United States)

    Zhang, Wen; Stack, Andrew G; Chen, Yongsheng

    2011-02-01

    To better understand environmental behaviors of nanoparticles (NPs), we used the atomic force microscopy (AFM) to measure interaction forces between E. coli cells and NPs immobilized on surfaces in an aqueous environment. The results showed that adhesion force strength was significantly influenced by particle size for both hematite (α-Fe(2)O(3)) and corundum (α-Al(2)O(3)) NPs whereas the effect on the repulsive force was not observed. The adhesion force decreased from 6.3±0.7nN to 0.8±0.4nN as hematite NPs increased from 26nm to 98nm in diameter. Corundum NPs exhibited a similar dependence of adhesion force on particle size. The Johnson-Kendall-Roberts (JKR) model was employed to estimate the contact area between E. coli cells and NPs, and based on the JKR model a new model that considers local effective contact area was developed. The prediction of the new model matched the size dependence of adhesion force in experimental results. Size effects on adhesion forces may originate from the difference in local effective contact areas as supported by our model. These findings provide fundamental information for interpreting the environmental behaviors and biological interactions of NPs, which barely have been addressed. Copyright © 2010 Elsevier B.V. All rights reserved.

  10. Study of Immobilization Procedure on Silver Nanolayers and Detection of Estrone with Diverged Beam Surface Plasmon Resonance (SPR Imaging

    Directory of Open Access Journals (Sweden)

    Ibrahim Abdulhalim

    2013-03-01

    Full Text Available An immobilization protocol was developed to attach receptors on smooth silver thin films. Dense and packed 11-mercaptoundecanoic acid (11-MUA was used to avoid uncontrolled sulfidization and harmful oxidation of silver nanolayers. N,N'-dicyclohexylcarbodiimide (DCC and N-hydroxysuccinimide (NHS were added to make the silver surfaces reactive. A comparative study was carried out with different immersion times of silver samples in 11-MUA solutions with different concentrations to find the optimum conditions for immobilization. The signals, during each step of the protocol, were analyzed with a refractometer based on the surface plasmon resonance (SPR effect and luminescence techniques. Molecular interactions at the surfaces between the probe and target at the surface nanolayer shift the SPR signal, thus indicating the presence of the substance. To demonstrate specific biosensing, rabbit anti-estrone polyclonal immunoglobulin G (IgG antibody was immobilized through a linker on 47 nm silver layer deposited on SF11 glass. At the final stage, the representative endocrine disruptor—estrone—was attached and detected in deionized water with a diverging beam SPR imaging sensor.

  11. Immobilization of gold nanoparticles on cell culture surfaces for safe and enhanced gold nanoparticle-mediated laser transfection

    Science.gov (United States)

    Kalies, Stefan; Heinemann, Dag; Schomaker, Markus; Gentemann, Lara; Meyer, Heiko; Ripken, Tammo

    2014-01-01

    Abstract. In comparison to standard transfection methods, gold nanoparticle-mediated laser transfection has proven to be a versatile alternative. This is based on its minor influence on cell viability and its high efficiency, especially for the delivery of small molecules like small interfering RNA. However, in order to transfer it to routine usage, a safety aspect is of major concern: The avoidance of nanoparticle uptake by the cells is desired. The immobilization of the gold nanoparticles on cell culture surfaces can address this issue. In this study, we achieved this by silanization of the appropriate surfaces and the binding of gold nanoparticles to them. Comparable perforation efficiencies to the previous approaches of gold nanoparticle-mediated laser transfection with free gold nanoparticles are demonstrated. The uptake of the immobilized particles by the cells is unlikely. Consequently, these investigations offer the possibility of bringing gold nanoparticle-mediated laser transfection closer to routine usage. PMID:25069006

  12. Immobilization of gold nanoparticles on cell culture surfaces for safe and enhanced gold nanoparticle-mediated laser transfection.

    Science.gov (United States)

    Kalies, Stefan; Heinemann, Dag; Schomaker, Markus; Gentemann, Lara; Meyer, Heiko; Ripken, Tammo

    2014-01-01

    In comparison to standard transfection methods, gold nanoparticle-mediated laser transfection has proven to be a versatile alternative. This is based on its minor influence on cell viability and its high efficiency, especially for the delivery of small molecules like small interfering RNA. However, in order to transfer it to routine usage, a safety aspect is of major concern: The avoidance of nanoparticle uptake by the cells is desired. The immobilization of the gold nanoparticles on cell culture surfaces can address this issue. In this study, we achieved this by silanization of the appropriate surfaces and the binding of gold nanoparticles to them. Comparable perforation efficiencies to the previous approaches of gold nanoparticle-mediated laser transfection with free gold nanoparticles are demonstrated. The uptake of the immobilized particles by the cells is unlikely. Consequently, these investigations offer the possibility of bringing gold nanoparticle-mediated laser transfection closer to routine usage.

  13. Development of thrombus-resistant and cell compatible crimped polyethylene terephthalate cardiovascular grafts using surface co-immobilized heparin and collagen

    Energy Technology Data Exchange (ETDEWEB)

    Al Meslmani, Bassam, E-mail: almeslmanib@yahoo.com [Department of Pharmaceutical Technology and Biopharmaceutics, Marburg University, Ketzerbach 63, 35037 Marburg (Germany); Mahmoud, Gihan, E-mail: mahmoudg@staff.uni-marburg.de [Department of Pharmaceutical Technology and Biopharmaceutics, Marburg University, Ketzerbach 63, 35037 Marburg (Germany); Department of Pharmaceutics and Industrial Pharmacy, Helwan University, Ain Helwan, 11795 Cairo (Egypt); Strehlow, Boris, E-mail: strehlo4@staff.uni-marburg.de [Department of Pharmaceutical Technology and Biopharmaceutics, Marburg University, Ketzerbach 63, 35037 Marburg (Germany); Mohr, Eva, E-mail: mohr@staff.uni-marburg.de [Department of Pharmaceutical Technology and Biopharmaceutics, Marburg University, Ketzerbach 63, 35037 Marburg (Germany); Leichtweiß, Thomas, E-mail: Thomas.leichtweiss@phys.chemie.uni-giessen.de [Institute of Physical Chemistry, Justus-Liebig-University, Heinrich-Buff-Ring 58, 35392 Giessen (Germany); Bakowsky, Udo, E-mail: ubakowsky@aol.com [Department of Pharmaceutical Technology and Biopharmaceutics, Marburg University, Ketzerbach 63, 35037 Marburg (Germany)

    2014-10-01

    Short-term patency of polyethylene terephthalate (PET) cardiovascular grafts is determined mainly by the inherent thrombogenicity and improper endothelialization following grafts implantation. The aim of the present study was to immobilize heparin to develop thrombus resistant grafts. Additionally, collagen was co-immobilized to enhance the host cell compatibility. The synthetic woven and knitted forms of crimped PET grafts were surface modified by Denier reduction to produce functional carboxyl groups. The produced groups were used as anchor sites for covalent immobilization of heparin or co-immobilization of heparin/collagen by the end-point method. The modified surface was characterized using Fourier transform infrared spectroscopy and X-ray photoelectron spectroscopy. The biological activity of immobilized molecules was investigated in vitro using direct blood coagulation test, and “platelet deposition under flow condition. Furthermore, the biocompatibility of modified grafts with host cells was assessed using L929 cell as model. All modified grafts showed significant resistance against fibrin and clot formation. The number of deposited platelets on heparin-immobilized woven and knitted grafts obviously decreased by 3 fold and 2.8 fold per unit surface area respectively, while the heparin/collagen co-immobilized grafts showed only a decrease by 1.7 and 1.8 fold compared to unmodified PET. Heparin-immobilized grafts reported no significant effect on L929 cells adhesion and growth (P > 0.05), conversely, collagen co-immobilization considerably increased cell adhesion almost ∼ 1.3 fold and 2 fold per unit surface area for woven and knitted grafts respectively. Our results emphasize that immobilization of heparin minimized the inherent thrombogenicity of the PET grafts. The simultaneous co-immobilization of collagen supported host cell adhesion and growth required for the grafts biocompatibility. - Highlight: • Heparin and collagen were co-immobilized on

  14. Self-assembled monolayers of 1-alkenes on oxidized platinum surfaces as platforms for immobilized enzymes for biosensing

    Energy Technology Data Exchange (ETDEWEB)

    Alonso, Jose Maria; Bielen, Abraham A.M. [Laboratory of Organic Chemistry, Wageningen University, Dreijenplein 8, 6703 HB, Wageningen (Netherlands); Olthuis, Wouter [BIOS Lab on a Chip Group, MESA+ and MIRA Institutes, University of Twente, P.O. Box 217, 7500 AE Enschede (Netherlands); Kengen, Servé W.M. [Laboratory of Microbiology, Wageningen University, 6703HB Wageningen (Netherlands); Zuilhof, Han, E-mail: han.zuilhof@wur.nl [Laboratory of Organic Chemistry, Wageningen University, Dreijenplein 8, 6703 HB, Wageningen (Netherlands); Department of Chemical and Materials Engineering, King Abdulaziz University, Jeddah 22254 (Saudi Arabia); Franssen, Maurice C.R., E-mail: maurice.franssen@wur.nl [Laboratory of Organic Chemistry, Wageningen University, Dreijenplein 8, 6703 HB, Wageningen (Netherlands)

    2016-10-15

    Highlights: • Three different oxidases are covalently attached to alkene based SAMs on PtOx. • Attached enzymes remain active and their activity is assessed by chronoamperometry. • Functionalized PtOx allows electron mediator free chronoamperometry measurements. • The thus formed enzyme electrodes are useful as biosensors for glucose and lactate. • Immobilization of human HAOX foresees in vivo lactate monitoring in humans. - Abstract: Alkene-based self-assembled monolayers grafted on oxidized Pt surfaces were used as a scaffold to covalently immobilize oxidase enzymes, with the aim to develop an amperometric biosensor platform. NH{sub 2}-terminated organic layers were functionalized with either aldehyde (CHO) or N-hydroxysuccinimide (NHS) ester-derived groups, to provide anchoring points for enzyme immobilization. The functionalized Pt surfaces were characterized by X-ray photoelectron spectroscopy (XPS), static water contact angle (CA), infrared reflection absorption spectroscopy (IRRAS) and atomic force microscopy (AFM). Glucose oxidase (GOX) was covalently attached to the functionalized Pt electrodes, either with or without additional glutaraldehyde crosslinking. The responses of the acquired sensors to glucose concentrations ranging from 0.5 to 100 mM were monitored by chronoamperometry. Furthermore, lactate oxidase (LOX) and human hydroxyacid oxidase (HAOX) were successfully immobilized onto the PtOx surface platform. The performance of the resulting lactate sensors was investigated for lactate concentrations ranging from 0.05 to 20 mM. The successful attachment of active enzymes (GOX, LOX and HAOX) on Pt electrodes demonstrates that covalently functionalized PtOx surfaces provide a universal platform for the development of oxidase enzyme-based sensors.

  15. Stabilization of a-glucosidase in organic solvents by immobilization on macroporous poly(GMA-co-EGDMA with different surface characteristics

    Directory of Open Access Journals (Sweden)

    RADIVOJE M. PRODANOVIC

    2006-04-01

    Full Text Available a-Glucosidase from baker’s yeast was immobilized on macroporous copolymers of ethylene glycol dimethacrylate and glycidyl methacrylate, poly(GMA-co-EGDMA, with various surface characteristics and pore sizes ranging from 44 nmto 270 nm. Immobilization was done by glutaraldehyde on the copolymer previously modified with 1,2-diaminoethane. The specific activity of the obtained immobilized enzyme varied from 27 to 81 U/g, depending on the employed copolymer. The half lives of the immobilized enzyme in cosolvents were influenced by the surface characteristics of the copolymer, ranging from 60 to 150 min in 35 % methanol and from 10 to 44 min in 45 % dimethyl sulphoxide (DMSO. The best stabilities were obtained when the enzyme was immobilized onto a copolymer having a pore size of 48 nm in methanol and 270 nm in DMSO.

  16. Immobilization of the iron on the surface of non-woven carbon fiber for use in a microbial fuel cell

    Directory of Open Access Journals (Sweden)

    Nichanan Phansroy

    2016-09-01

    Full Text Available Abstract Iron particles were immobilized onto non-woven carbon fiber via electroplating for use in a microbial fuel cell (MFC. Electroplating was performed under an applied voltage at a current of 0.2 A for 5, 10, and 15 min. The scanning electron microscope (SEM observations show that 5 min was not adequate for the particles to be immobilized, whereas 10 and 15 min of electroplating resulted in an adequate number of particles on the surface. To evaluate the strength of the binding of iron via electroplating on the surface of the fiber, the samples were washed with pure water and observed using an SEM. The 10 min electroplated sample has a larger surface area, which is suitable for the MFC anode, than the 15 min electroplated sample. According to X-ray photoelectron spectroscopy and X-ray diffraction analysis, the peaks corresponded to those of Fe2O3, and the sample dipped into tannic acid shows the peaks of Fe3O4. The amount of biofilm of Shewanella oneidensis MR-1 was evaluated using crystal violet staining, and living bacteria were counted as colony forming units. Electroplated Fe2O3 and Fe3O4 were found to be effective for producing biofilm and immobilizing S. oneidensis MR-1.

  17. Surface modification of polyacrylonitrile fiber for immobilization of antibodies and detection of analyte

    International Nuclear Information System (INIS)

    Jain, Swati; Chattopadhyay, Sruti; Jackeray, Richa; Singh, Harpal

    2009-01-01

    Pendent nitrile groups of multifilamentous polyacrylonitrile (PAN) fibers were reduced to amino groups using lithium aluminum hydride for different time of reduction and amine content was estimated by performing acid-base titrations. Attenuated total reflection-fourier transform infrared spectroscopy (ATR-FTIR) and Differential Scanning Calorimetry (DSC) were used for the characterization of the generated amino groups and thermal properties of the reduced fibers, respectively. The surface morphology of the fibers after reduction and immobilization was characterized using Scanning Electron Microscope (SEM). The newly formed amino groups of the fibers were activated by using glutaraldehyde for the covalent linking of Goat anti-Rabbit IgG-HRP (GAR-HRP) antibody enzyme conjugate. Modified PAN fibers were evaluated as a matrix for sandwich ELISA by using Goat anti-Rabbit antibody (GAR-IgG), Rabbit anti-Goat (RAG-IgG) as analyte and enzyme conjugate GAR-HRP. The fibers reduced for 24 h were able to detect the analyte RAG-IgG at a concentration as low as 3.75 ng mL -1 with 12% skimmed milk as blocking reagent for the optimized concentration of primary antibody GAR-IgG 3 μg mL -1 and peroxidase conjugate GAR-HRP dilution of 8000 fold. The sensitivity, specificity and reproducibility of the developed immunoassay was further established with antibodies present in human blood using Rabbit anti-Human (RAH-IgG) antibody and the corresponding HRP enzyme conjugate. As low as 0.1 μL of human blood was sufficient to perform the assay with the modified fibers.

  18. Surface modification of polyacrylonitrile fiber for immobilization of antibodies and detection of analyte.

    Science.gov (United States)

    Jain, Swati; Chattopadhyay, Sruti; Jackeray, Richa; Singh, Harpal

    2009-11-10

    Pendent nitrile groups of multifilamentous polyacrylonitrile (PAN) fibers were reduced to amino groups using lithium aluminum hydride for different time of reduction and amine content was estimated by performing acid-base titrations. Attenuated total reflection-fourier transform infrared spectroscopy (ATR-FTIR) and Differential Scanning Calorimetry (DSC) were used for the characterization of the generated amino groups and thermal properties of the reduced fibers, respectively. The surface morphology of the fibers after reduction and immobilization was characterized using Scanning Electron Microscope (SEM). The newly formed amino groups of the fibers were activated by using glutaraldehyde for the covalent linking of Goat anti-Rabbit IgG-HRP (GAR-HRP) antibody enzyme conjugate. Modified PAN fibers were evaluated as a matrix for sandwich ELISA by using Goat anti-Rabbit antibody (GAR-IgG), Rabbit anti-Goat (RAG-IgG) as analyte and enzyme conjugate GAR-HRP. The fibers reduced for 24h were able to detect the analyte RAG-IgG at a concentration as low as 3.75ng mL(-1) with 12% skimmed milk as blocking reagent for the optimized concentration of primary antibody GAR-IgG 3microg mL(-1) and peroxidase conjugate GAR-HRP dilution of 8000 fold. The sensitivity, specificity and reproducibility of the developed immunoassay was further established with antibodies present in human blood using Rabbit anti-Human (RAH-IgG) antibody and the corresponding HRP enzyme conjugate. As low as 0.1microL of human blood was sufficient to perform the assay with the modified fibers.

  19. Surface modification of polyacrylonitrile fiber for immobilization of antibodies and detection of analyte

    Energy Technology Data Exchange (ETDEWEB)

    Jain, Swati, E-mail: swatijain.iitd@gmail.com [Center for Biomedical Engineering, Indian Institute of Technology, New Delhi, 110016 (India); Chattopadhyay, Sruti, E-mail: srutic@hotmail.com [Center for Biomedical Engineering, Indian Institute of Technology, New Delhi, 110016 (India); Jackeray, Richa, E-mail: richajackeray.iitd@gmail.com [Center for Biomedical Engineering, Indian Institute of Technology, New Delhi, 110016 (India); Singh, Harpal, E-mail: harpal2000@yahoo.com [Center for Biomedical Engineering, Indian Institute of Technology, New Delhi, 110016 (India)

    2009-11-10

    Pendent nitrile groups of multifilamentous polyacrylonitrile (PAN) fibers were reduced to amino groups using lithium aluminum hydride for different time of reduction and amine content was estimated by performing acid-base titrations. Attenuated total reflection-fourier transform infrared spectroscopy (ATR-FTIR) and Differential Scanning Calorimetry (DSC) were used for the characterization of the generated amino groups and thermal properties of the reduced fibers, respectively. The surface morphology of the fibers after reduction and immobilization was characterized using Scanning Electron Microscope (SEM). The newly formed amino groups of the fibers were activated by using glutaraldehyde for the covalent linking of Goat anti-Rabbit IgG-HRP (GAR-HRP) antibody enzyme conjugate. Modified PAN fibers were evaluated as a matrix for sandwich ELISA by using Goat anti-Rabbit antibody (GAR-IgG), Rabbit anti-Goat (RAG-IgG) as analyte and enzyme conjugate GAR-HRP. The fibers reduced for 24 h were able to detect the analyte RAG-IgG at a concentration as low as 3.75 ng mL{sup -1} with 12% skimmed milk as blocking reagent for the optimized concentration of primary antibody GAR-IgG 3 {mu}g mL{sup -1} and peroxidase conjugate GAR-HRP dilution of 8000 fold. The sensitivity, specificity and reproducibility of the developed immunoassay was further established with antibodies present in human blood using Rabbit anti-Human (RAH-IgG) antibody and the corresponding HRP enzyme conjugate. As low as 0.1 {mu}L of human blood was sufficient to perform the assay with the modified fibers.

  20. IMMOBILIZATION OF MICROALGAE ON THE SURFACE OF NEW CROSS-LINKED POLYETHYLENIMINE-BASED SORBENTS.

    Science.gov (United States)

    Vasilieva, Svetlana; Shibzukhova, Karina; Morozov, Alexey; Solovchenko, Alexei; Bessonov, Ivan; Kopitsyna, Maria; Lukyanov, Alexander; Chekanov, Konstantin; Lobakova, Elena

    2018-04-11

    We report on the use of the polyethylenimine-based (PEI) sorbents for immobilization and harvesting of microalgae (MA) cells. Specific materials assessed were porous solid polymers from highly-branched PEI synthesized by cross-linking with epichlorohydrin (ECH) or diethylene glycol diglycidyl ether (DGDE). We estimated the effect of PEI/cross-linker ratio on the MA attachment and biocompatibility of the sorbents with the MA cells. A decrease in the cross-linker percentage resulted in the enhancement of the immobilization efficiency but impaired the cell viability as was manifested by inhibition of the photosynthetic activity of the MA cells. The rate of Chlorella vulgaris cell attachment to the sorbents with ECH was faster as compared to that of the PEI-DGDE-based polymers. The cells immobilized on the PEI-ECH sorbents showed a more profound decline in their viability (assessed via photosynthetic activity). The sorbents with 60% of DGDE were characterized by high immobilization efficiency. These sorbents supported a prolonged cultivation of the immobilized MA without impairing their viability and metabolic activity. We conclude that the sorbents with a lower percentage of DGDE (<30%) and sorbents with ECH are suitable for harvesting of the MA cells intended for immediate downstream processing, potentially without the cell desorption. To the best of our knowledge, this is the first report on successful application of PEI-based sorbents in microalgal biotechnology. Copyright © 2018 Elsevier B.V. All rights reserved.

  1. Covalent and Oriented Surface Immobilization of Antibody Using Photoactivatable Antibody Fc-Binding Protein Expressed in Escherichia coli.

    Science.gov (United States)

    Lee, Yeolin; Jeong, Jiyun; Lee, Gabi; Moon, Jeong Hee; Lee, Myung Kyu

    2016-10-04

    Fc-specific antibody binding proteins (FcBPs) with the minimal domain of protein G are widely used for immobilization of well-oriented antibodies onto solid surfaces, but the noncovalently bound antibodies to FcBPs are unstable in sera containing large amounts of antibodies. Here we report novel photoactivatable FcBPs with photomethionine (pMet) expressed in E. coli, which induce Fc-specific photo-cross-linking with antibodies upon UV irradiation. Unfortunately, pMet did not support protein expression in the native E. coli system, and therefore we also developed an engineered methionyl tRNA synthetase (MRS5m). Coexpression of MRS5m proteins successfully induced photoactivatable FcBP overexpression in methionine-auxotroph E. coli cells. The photoactivatable FcBPs could be easily immobilized on beads and slides via their N-terminal cysteine residues and 6xHis tag. The antibodies photo-cross-linked onto the photoactivatable FcBP-beads were resistant from serum-antibody mediated dissociation and efficiently captured antigens in human sera. Furthermore, photo-cross-linked antibody arrays prepared using this system allowed sensitive detection of antigens in human sera by sandwich immunoassay. The photoactivatable FcBPs will be widely applicable for well-oriented antibody immobilization on various surfaces of microfluidic chips, glass slides, and nanobeads, which are required for development of sensitive immunosensors.

  2. Enzymes immobilized in Langmuir-Blodgett films: Why determining the surface properties in Langmuir monolayer is important?

    Directory of Open Access Journals (Sweden)

    LUCIANO CASELI

    2018-01-01

    Full Text Available ABSTRACT In this review we discuss about the immobilization of enzymes in Langmuir-Blodgett films in order to determine the catalytic properties of these biomacromolecules when adsorbed on solid supports. Usually, the conformation of enzymes depends on the environmental conditions imposed to them, including the chemical composition of the matrix, and the morphology and thickness of the film. In this review, we show an outline of manuscripts that report the immobilization of enzymes as LB films since the 1980’s, and also some examples of how the surface properties of the floating monolayer prepared previously to the transfer to the solid support are important to determine the efficiency of the resulting device.

  3. The effect of the shape of single, sub-ms voltage pulses on the rates of surface immobilization and hybridization of DNA

    International Nuclear Information System (INIS)

    Cabeca, R; Rodrigues, M; Chu, V; Conde, J P; Prazeres, D M F

    2009-01-01

    Electric fields generated by single square and sinusoidal voltage pulses with amplitudes below 2 V were used to assist the covalent immobilization of single-stranded, thiolated DNA probes, onto a chemically functionalized SiO 2 surface and to assist the specific hybridization of single-stranded DNA targets with immobilized complementary probes. The single-stranded immobilized DNA probes were either covalently immobilized (chemisorption) or electrostatically adsorbed (physisorption) to a chemically functionalized surface. Comparing the speed of electric field assisted immobilization and hybridization with the corresponding control reactions (without electric field), an increase of several orders of magnitude is observed, with the reaction timescaled down from 1 to 2 h to a range between 100 ns and 1 ms. The influence of the shape of the voltage pulse (square versus sinusoidal) and its duration were studied for both immobilization and hybridization reactions. The results show that pulsed electric fields are a useful tool to achieve temporal and spatial control of surface immobilization and hybridization reactions of DNA.

  4. Soluble and immobilized graphene oxide activates complement system differently dependent on surface oxidation state

    DEFF Research Database (Denmark)

    Wibroe, Peter Popp; Petersen, Søren Vermehren; Bovet, Nicolas Emile

    2016-01-01

    Graphene oxide (GO) is believed to become applicable in biomedical products and medicine, thereby necessitating appropriate safety evaluation dependent on their applications and the route of administration. We have examined the effect of GO form (in solution versus immobilized) and oxidation stat...

  5. Surface Immobilization of Human Arginase-1 with an Engineered Ice Nucleation Protein Display System in E. coli.

    Directory of Open Access Journals (Sweden)

    Zhen Zhang

    Full Text Available Ice nucleation protein (INP is frequently used as a surface anchor for protein display in gram-negative bacteria. Here, MalE and TorA signal peptides, and three charged polypeptides, 6×Lys, 6×Glu and 6×Asp, were anchored to the N-terminus of truncated INP (InaK-N to improve its surface display efficiency for human Arginase1 (ARG1. Our results indicated that the TorA signal peptide increased the surface translocation of non-protein fused InaK-N and human ARG1 fused InaK-N (InaK-N/ARG1 by 80.7% and 122.4%, respectively. Comparably, the MalE signal peptide decreased the display efficiencies of both the non-protein fused InaK-N and InaK-N/ARG1. Our results also suggested that the 6×Lys polypeptide significantly increased the surface display efficiency of K6-InaK-N/ARG1 by almost 2-fold, while also practically abolishing the surface translocation of non-protein fused InaK-N, indicating the interesting roles of charged polypeptides in bacteria surface display systems. Cell surface-immobilized K6-InaK-N/ARG1 presented an arginase activity of 10.7 U/OD600 under the optimized conditions of 40°C, pH 10.0 and 1 mM Mn2+, which could convert more than 95% of L-Arginine (L-Arg to L-Ornithine (L-Orn in 16 hours. The engineered InaK-Ns expanded the INP surface display system, which aided in the surface immobilization of human ARG1 in E. coli cells.

  6. New insights into the effectiveness of alpha-amylase enzyme presentation on the Bacillus subtilis spore surface by adsorption and covalent immobilization.

    Science.gov (United States)

    Gashtasbi, Fatemeh; Ahmadian, Gholamreza; Noghabi, Kambiz Akbari

    2014-10-01

    Most of the studies in the field of enzyme immobilization have sought to develop a simple, efficient and cost-effective immobilization system. In this study, probiotic Bacillus spores were used as a matrix for enzyme immobilization, because of their inherent resistance to extreme temperatures, UV irradiation, solvents and drying. Above all, their preparation is cost-effective. The alpha-amylase enzyme was immobilized on the spore surface by the covalent and adsorption methods. For the covalent method, 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) and N hydroxysulfosuccinimide (NHS) were used. The maximum concentration of the alpha-amylase immobilized by the two methods onto the spore surface was 360 μg/1.2×10(11) spore. However, maximum activity was achieved at an enzyme concentration of approximately 60 μg/.4×10(10), corresponding to an estimated activity of 8×10(3) IU mg(-1)/1.2×10(11) spore for covalent immobilization and 8.53×10(3) for the adsorption method. After washing the enzyme with 1M NaCl and 0.5% Triton X-100, the enzyme immobilization yield was estimated to be 77% and 20.07% for the covalent and adsorption methods, respectively. The alpha-amylase immobilized by both methods, displayed improved activity in the basic pH range. The optimum pH for the free enzyme was 5 while it shifted to 8 for the immobilized enzyme. The optimum temperatures for the free and immobilized enzymes were 60 °C and 80 °C, respectively. The covalently-immobilized alpha-amylase retained 65% of its initial activity, even after 10 times of recycling. The Km and Vmax values were determined by the GraphPad Prism software, which showed that the Vmax value decreased moderately after immobilization. This is the first study which reports the covalent immobilization of an enzyme on the spore surface. Copyright © 2014 Elsevier Inc. All rights reserved.

  7. Immobilization of ionophore and surface characterization studies of the titanium(III) ion in a PVC-membrane sensor.

    Science.gov (United States)

    Rezayi, Majid; Heng, Lee Yook; Kassim, Anuar; Ahmadzadeh, Saeid; Abdollahi, Yadollah; Jahangirian, Hossein

    2012-01-01

    Novel ionophores comprising various hydroxide and amine structures were immobilized onto poly(vinyl chloride) (PVC) matrices, and these were examined to determine Ti(III) selectivity. To predict the selectivity of Ti(III), a PVC membrane was used to investigate the binding of Ti(III) to c-methylcalix[4]resorcinarene (CMCR). The study showed that the chelating ligand, CMCR, was coordinated selectively to Ti(III) at eight coordination sites involving the oxygen atoms at the interface of the membrane/solution. The membrane was prepared, based on CMCR as an ionophore, sodium tetrakis(4-fluorophenyl) borate (NaTFPB) as a lipophilic ionic additive, and dioctylphthalate (DOP) as a plasticizer. The immobilization of the ionophore and surface characterization studies revealed that the performance of CMCR-immobilized PVC was equivalent to that of mobile ionophores in supported liquid membranes (SLMs). The strengths of the ion-ionophore (CMCR-Ti(OH)(OH(2))(5) (2+)) interactions and the role of ionophores on membranes were studied via UV-Vis, Fourier transform infrared spectroscopy (FT-IR), scanning electron microscopy (SEM) and and X-ray diffraction (XRD).

  8. Cobalt(II phthalocyanine bonded to 3-n-propylimidazole immobilized on silica gel surface: preparation and electrochemical properties

    Directory of Open Access Journals (Sweden)

    Fujiwara Sergio T.

    1999-01-01

    Full Text Available Co-Phthalocyanine complex was immobilized on 3-n-propylimidazole groups grafted on a porous SiO2 surface (specific surface area S BET = 500 m² g-1 and efficiently electrocatalyzed the oxalic acid oxidation on a carbon paste electrode surface made of this material. Intermolecular interactions of the complex species which can normally interfere in the redox process practically are not observed in the present case because of a low average surface density, delta = 4.7 x 10-13 mol cm-2 (delta = Nf/S BET, where Nf is the amount of adsorbed Co-phtalocyanine per gram of modified silica gel of the complex species material prepared. The linear response of the electrode to oxalic acid concentration, between 6.5 x 10-4 and 3.2 x 10-3 mol L-1, associated with its high chemical stability makes the covalently immobilized Co-phtalocyanine complex material very attractive in preparing a new class of chemical sensors.

  9. Ultra-small rhenium nanoparticles immobilized on DNA scaffolds: An excellent material for surface enhanced Raman scattering and catalysis studies.

    Science.gov (United States)

    Anantharaj, S; Sakthikumar, K; Elangovan, Ayyapan; Ravi, G; Karthik, T; Kundu, Subrata

    2016-12-01

    Highly Sensitive and ultra-small Rhenium (Re) metal nanoparticles (NPs) were successfully stabilized in water by the staging and fencing action of the versatile biomolecule DNA that resulted in two distinct aggregated chain-like morphologies with average grain sizes of 1.1±0.1nm and 0.7±0.1nm for the very first time within a minute of reaction time. Re NPs are formed by the borohydride reduction of ammonium perrhenate (NH4ReO4) in the presence of DNA at room temperature (RT) under stirring. The morphologies were controlled by carefully monitoring the molar ratio of NH4ReO4 and DNA. The synthesized material was employed in two potential applications: as a substrate for surface enhanced Raman scattering (SERS) studies and as a catalyst for the reduction of aromatic nitro compounds. SERS study was carried out by taking methylene blue (MB) as the probe and the highest SERS enhancement factor (EF) of 2.07×10(7) was found for the aggregated chain-like having average grain size of 0.7±0.1nm. Catalytic reduction of 4-nitro phenol (4-NP), 2-nitro phenol (2-NP) and 4-nitroaniline (4-NA) with a rate constant value of 6×10(-2)min(-1), 33.83×10(-2)min(-1) and 37.4×10(-2)min(-1) have testified the excellent catalytic performance of our Re NPs immobilized on DNA. The overall study have revealed the capability of DNA in stabilizing the highly reactive Re metal at nanoscale and made them applicable in practice. The present route can also be extended to prepare one dimensional (1-D), self-assembled NPs of other reactive metals, mixed metals or even metal oxides for specific applications in water based solutions. Copyright © 2016 Elsevier Inc. All rights reserved.

  10. Supramolecular protein immobilization on lipid bilayers

    NARCIS (Netherlands)

    Bosmans, R.P.G.; Hendriksen, W.E.; Verheijden, Mark Lloyd; Eelkema, R.; Jonkheijm, Pascal; van Esch, J.H.; Brunsveld, Luc

    2015-01-01

    Protein immobilization on surfaces, and on lipid bilayers specifically, has great potential in biomolecular and biotechnological research. Of current special interest is the immobilization of proteins using supramolecular noncovalent interactions. This allows for a reversible immobilization and

  11. Immobilization, hybridization, and oxidation of synthetic DNA on gold surface: Electron transfer investigated by electrochemistry and scanning tunneling microscopy

    Energy Technology Data Exchange (ETDEWEB)

    McEwen, Gerald D.; Chen Fan [Biological Engineering Program, Department of Biological and Irrigation Engineering, Utah State University, 4105 Old Main Hill, Logan, UT 84322-4105 (United States); Zhou Anhong, E-mail: Anhong.Zhou@usu.edu [Biological Engineering Program, Department of Biological and Irrigation Engineering, Utah State University, 4105 Old Main Hill, Logan, UT 84322-4105 (United States)

    2009-06-08

    Fundamental understanding of interfacial electron transfer (ET) among electrolyte/DNA/solid-surface will facilitate the design for electrical detection of DNA molecules. In this report, the electron transfer characteristics of synthetic DNA (sequence from pathogenic Cryptosporidium parvum) self-assembled on a gold surface was electrochemically studied. The effects of immobilization order on the interface ET related parameters such as diffusion coefficient (D{sub 0}), surface coverage ({theta}{sub R}), and monolayer thickness (d{sub i}) were determined by cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). DNA surface density ({Gamma}{sub DNA}) was determined by the integration of the charge of the electro-oxidation current peaks during the initial cyclic voltammetry scans. It was found that the DNA surface densities at different modifications followed the order: {Gamma}{sub DNA} (dsS-DNA/Au) > {Gamma}{sub DNA} (MCH/dsS-DNA/Au) > {Gamma}{sub DNA} (dsS-DNA/MCH/Au). It was also revealed that the electro-oxidation of the DNA modified gold surface would involve the oxidation of nucleotides (guanine and adenine) with a 5.51 electron transfer mechanism and the oxidative desorption of DNA and MCH molecules by a 3 electron transfer mechanism. STM topography and current image analysis indicated that the surface conductivity after each surface modification followed the order: dsS-DNA/Au < MCH/dsS-DNA/Au < oxidized MCH/dsS-DNA/Au < Hoechst/oxidized MCH/dsS-DNA/Au. The results from this study suggested a combination of variations in immobilization order may provide an alternative approach for the optimization of DNA hybridization and the further development for electrical detection of DNA.

  12. Surface properties and in vitro analyses of immobilized chitosan onto polypropylene non-woven fabric surface using antenna-coupling microwave plasma.

    Science.gov (United States)

    Tyan, Yu-Chang; Liao, Jiunn-Der; Lin, Shu-Ping

    2003-09-01

    Antenna coupling microwave plasma enables a highly efficient and oxidative treatment of the outermost surface of polypropylene (PP) non-woven fabric within a short time period. Subsequently, grafting copolymerization with acrylic acid (AAc) makes the plasma-treated fabric durably hydrophilic and excellent in water absorbency. With high grafting density and strong water affinity, the pAAc-grafted fabric greatly becomes feasible as an intensive absorbent and as a support to promote chitosan-immobilization through amide bonds. Experimental result demonstrated that surface analyses by FTIR-ATR have shown that R-CONH-R', amide binding were emerged between pAAc and chitosan. The XPS measurements on C(1s) 286.0 eV (C-OH), 286.5 eV (C-N) and 288.1 eV (O=C-NH) also could be found. Bioactivity assessments on the chitosan-immobilized surfaces were anticipated by activated partial thromboplastin time (aPTT), thrombin time (TT), and fibrinogen concentration. By means of cell counter we counted the ratio of blood cell adhesion on the modified fabric matrix. After human plasma incubated with the chitosan-immobilized PP fabrics, the required time for aPTT and blood cell adhesion increased significantly, while fibrinogen concentration and TT did not change. Due to the capability of anticoagulation and cell adhesion, the chitosan-immobilized PP fabric can be used as the substrate for cell culturing and then developed the wound-dressing substitute for second-degree burn.

  13. Tyrosinase-catalyzed site-specific immobilization of engineered C-phycocyanin to surface

    Science.gov (United States)

    Faccio, Greta; Kämpf, Michael M.; Piatti, Chiara; Thöny-Meyer, Linda; Richter, Michael

    2014-06-01

    Enzymatic crosslinking of proteins is often limited by the steric availability of the target residues, as of tyrosyl side chains in the case of tyrosinase. Carrying an N-terminal peptide-tag containing two tyrosine residues, the fluorescent protein C-phycocyanin HisCPC from Synechocystis sp. PCC6803 was crosslinked to fluorescent high-molecular weight forms with tyrosinase. Crosslinking with tyrosinase in the presence of L-tyrosine produced non fluorescent high-molecular weight products. Incubated in the presence of tyrosinase, HisCPC could also be immobilized to amino-modified polystyrene beads thus conferring a blue fluorescence. Crosslinking and immobilization were site-specific as both processes required the presence of the N-terminal peptide in HisCPC.

  14. Supply and cost factors for metals in the Canadian nuclear fuel waste immobilization program

    International Nuclear Information System (INIS)

    McConnell, D.B.

    1982-11-01

    Estimates have been made of the demand for immobilization containers to accommodate the irradiated fuel bundles arising from Canadian nuclear generating stations to the year 2020. The resulting estimates for container shells and container-filling alloys were compared to estimates for Canadian and Western World production of the candiate metals. The results indicate that, among the container shell metals, supply difficulties might arise only for Grade 7 titanium. Among the filling metals, only lead-antimony alloy might present supply problems. Current cost figures for plate made of each shell metal, and bulk quantities of filling metals, were compared. Materials costs would be least for a supported shell of stainless steel, followed by copper, titanium alloys Grades 2, 12 and 7, and Inconel 625. Aluminum-silicon is the lowest-cost filling matrix, followed by zinc, lead, and lead-antimony. Container durability, vault conditions, groundwater composition and other factors may play an overriding role in the final selection of materials for container construction

  15. SU-E-T-348: Effect of Treatment Table and Immobilization Devices On Surface Dose When Using a GRID Technique

    Energy Technology Data Exchange (ETDEWEB)

    Gajdos, S; Donaghue, J [Akron General Medical Center, Akron, OH (United States)

    2015-06-15

    Purpose: To determine the increase of surface dose of MLC-designed GRID therapy in the presence of immobilization devices and treatment table. Methods: To create a GRID field, our facility utilizes an MLC consisting of four millimeter wide leaves. The field is designed to have aperture sizes of 0.8 cm X 0.8 cm with inter-aperture distance of 3.2 cm. Gafchromic EBT3 film was placed between the surface of a solid water phantom and the immobilization device. The treatment table was also present within the beam path. The devices consist of carbon fiber exterior shell. A piece of film was also placed at maximal depth for the photon energy of 10 MV. Image files were converted to dose per a calibration curve based on the selected red channel. The surface dose to maximum dose was established by comparing the ratio of seven centrally located aperture regions-of-interest and four adjacent inter-aperture regions-of-interest were measured with the available software tools. Results: With no devices present in beam path, the ratio of surface dose to maximum dose was 11.5% ± 0.3% for aperture region and 7.0% ± 0.1% for inter-aperture region. When devices are present, the ratio of surface dose to maximum dose was 45.2% ± 0.5% and 33.8% ± 1.1%, respectively. Due to the presence of devices, the surface dose increases in aperture region by 3.8 times or in the inter-aperture region by 4.7 times. Conclusion: The purpose of using GRID technique is to deliver a single fractional dose in range of 15–20 Gy to a bulky lesion while also preserving skin tolerance. The increase of surface dose due to devices placed in beam path may increase the chance of skin toxicity in GRID therapy. Care should be used to determine best manageable patient immobilization while considering skin dose especially for posteriorly located lesions.

  16. Protein immobilization on the surface of polydimethylsiloxane and polymethyl methacrylate microfluidic devices.

    Science.gov (United States)

    Khnouf, Ruba; Karasneh, Dina; Albiss, Borhan Aldeen

    2016-02-01

    PDMS and PMMA are two of the most used polymers in the fabrication of lab-on-chip or microfluidic devices. In order to use these polymers in biological applications, it is sometimes essential to be able to bind biomolecules such as proteins and DNA to the surface of these materials. In this work, we have evaluated a number of processes that have been developed to bind protein to PDMS surfaces which include passive adsorption, passive adsorption with glutaraldehyde cross-linking, (3-aminopropyl) triethoxysilane functionalization followed by glutaraldehyde or 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride cross-linkers. It has been shown that the latter technique--using 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride--results in more than twice the bonding of protein to the surface of PDMS microchannels than proteins binding passively. We have also evaluated a few techniques that have been tested for the functionalization of PMMA microchannels where we have found that the use of polyethyleneimine (PEI) has led to the strongest protein-PMMA microchannel bond. We finally demonstrated the effect of PDMS curing methodology on protein adsorption to its surface, and showed that increased curing time is the factor that reduces passive adsorption the most. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  17. Enhancing oil removal from water by immobilizing multi-wall carbon nanotubes on the surface of polyurethane foam.

    Science.gov (United States)

    Keshavarz, Alireza; Zilouei, Hamid; Abdolmaleki, Amir; Asadinezhad, Ahmad

    2015-07-01

    A surface modification method was carried out to enhance the light crude oil sorption capacity of polyurethane foam (PUF) through immobilization of multi-walled carbon nanotube (MWCNT) on the foam surface at various concentrations. The developed sorbent was characterized using scanning electron microscopy, Fourier transform infrared spectroscopy, thermogravimetric analysis, and tensile elongation test. The results obtained from thermogravimetric and tensile elongation tests showed the improvement of thermal and mechanical resistance of surface-modified foam. The experimental data also revealed that the immobilization of MWCNT on PUF surface enhanced the sorption capacity of light crude oil and reduced water sorption. The highest oil removal capacity was obtained for 1 wt% MWCNT on PUF surface which was 21.44% enhancement in light crude oil sorption compared to the blank PUF. The reusability of surface modified PUF was determined through four cycles of chemical regeneration using petroleum ether. The adsorption of light crude oil with 30 g initial mass showed that 85.45% of the initial oil sorption capacity of this modified sorbent was remained after four regeneration cycles. Equilibrium isotherms for adsorption of oil were analyzed by the Freundlich, Langmuir, Temkin, and Redlich-Peterson models through linear and non-linear regression methods. Results of equilibrium revealed that Langmuir isotherm is the best fitting model and non-linear method is a more accurate way to predict the parameters involved in the isotherms. The overall findings suggested the promising potentials of the developed sorbent in order to be efficiently used in large-scale oil spill cleanup. Copyright © 2015 Elsevier Ltd. All rights reserved.

  18. The immobilization of a direct thrombin inhibitor to a polyurethane as a nonthrombogenic surface coating for extracorporeal circulation

    Science.gov (United States)

    Yu, Jane; Brisbois, Elizabeth; Handa, Hitesh; Annich, Gail; Meyerhoff, Mark; Bartlett, Robert; Major, Terry

    2016-01-01

    A biomaterial with both antithrombin and antiplatelet properties is the ideal surface for use in extracorporeal circulation (ECC) as it targets both fibrin generation and platelet adhesion. A hemocompatible surface coating avoids the need for systemic anticoagulation by providing a local anticoagulant effect at the polymer-blood interface. Previous work has demonstrated the potential use of argatroban, a direct thrombin inhibitor, as a nonthrombogenic material for extracorporeal devices. The work reported here focuses on the characterization of argatroban linked to a polyurethane-silicone polymer, CarboSil®. Chemical immobilization, the amount of argatroban, incubation times, and saturation point were evaluated to achieve maximal antithrombin activity at the polymer surface. Cross-linked polymer coatings reacted with 10 and 30 µmole of argatroban were prepared and tested. These coatings resulted in argatroban activity levels of 0.131 µM and 0.446 µM, respectively. After refining the cross-linking process, argatroban activity increased by approximately 3.6 fold. Maintenance of activity and leaching from the polymer surface were also evaluated. Using the refined process for linking argatroban to polymer, the resulting polymer was applied as a surface coating to the inner lumen of poly(vinyl chloride) ECC circuit tubing and its antithrombin effect evaluated using a 4 h rabbit ECC model. Following 4 h of blood exposure, the argatroban circuit demonstrated significantly less thrombus formation compared to the control CarboSil® coating with a 4.1 cm2 thrombus average area for the control coating compared to 1.2 cm2 for the argatroban coating (n=4). There was no significant change in thrombin time from baseline in plasma from animals in which the argatroban coated circuit was used, with a thrombin time of 16.2 s at t=0 and 14.5 s after 4 h. These results demonstrate the potential efficacy of immobilized argatroban as a hemocompatible biomaterial for extracorporeal

  19. Cell-surface interactions involving immobilized magnetite nanoparticles on flat magnetic substrates.

    Science.gov (United States)

    Loichen, Juliane; Hartmann, Uwe

    2009-09-01

    A new method to affect cells by cell-surface interaction is introduced. Biocompatible magnetic nanobeads are deposited onto a biocompatible magnetic thin layer. The particles are composed of small magnetite crystals embedded in a matrix which can be functionalized by different molecules, proteins or growth factors. The magnetic interaction between surface and beads prevents endocytosis if the setup is utilized for cell culturing. The force acting between particles and magnetic layer is calculated by a magnetostatic approach. Biocompatibility is ensured by using garnet layers which turned out to be nontoxic and stable under culturing conditions. The garnet thin films exhibit spatially and temporally variable magnetic domain configurations in changing external magnetic fields and depending on their thermal pretreatment. Several patterns and bead deposition methods as well as the cell-surface interactions were analyzed. In some cases the cells show directed growth. Theoretical considerations explaining particular cell behavior on this magnetic material involve calculations of cell growth on elastic substrates and bending of cell membranes.

  20. Cross-Linked ZnO Nanowalls Immobilized onto Bamboo Surface and Their Use as Recyclable Photocatalysts

    Directory of Open Access Journals (Sweden)

    Chunde Jin

    2014-01-01

    Full Text Available A novel recyclable photocatalyst was fabricated by hydrothermal method to immobilize the cross-linked ZnO nanowalls on the bamboo surface. The resultant samples were characterized by using scanning electron microscopy (SEM, X-ray diffraction (XRD, energy dispersive spectroscopy (EDS, and Fourier transformation infrared (FTIR techniques. FTIR spectra demonstrated that the cross-linked wurtzite ZnO nanowalls and bamboo surface were interconnected with each other by hydrogen bonds. Meanwhile, the cross-linked ZnO nanowalls modified bamboo (CZNB presented a superior photocatalytic ability and could be recycled at least 3 times with a photocatalytic efficiency up to 70%. The current research provides a new opportunity for the development of a portable and recycled biomass-based photocatalysts which can be an efficiently degraded pollutant solution and reused several times.

  1. Modulation of Protein Fouling and Interfacial Properties at Carbon Surfaces via Immobilization of Glycans Using Aryldiazonium Chemistry

    Science.gov (United States)

    Zen, Federico; Angione, M. Daniela; Behan, James A.; Cullen, Ronan J.; Duff, Thomas; Vasconcelos, Joana M.; Scanlan, Eoin M.; Colavita, Paula E.

    2016-01-01

    Carbon materials and nanomaterials are of great interest for biological applications such as implantable devices and nanoparticle vectors, however, to realize their potential it is critical to control formation and composition of the protein corona in biological media. In this work, protein adsorption studies were carried out at carbon surfaces functionalized with aryldiazonium layers bearing mono- and di-saccharide glycosides. Surface IR reflectance absorption spectroscopy and quartz crystal microbalance were used to study adsorption of albumin, lysozyme and fibrinogen. Protein adsorption was found to decrease by 30–90% with respect to bare carbon surfaces; notably, enhanced rejection was observed in the case of the tested di-saccharide vs. simple mono-saccharides for near-physiological protein concentration values. ζ-potential measurements revealed that aryldiazonium chemistry results in the immobilization of phenylglycosides without a change in surface charge density, which is known to be important for protein adsorption. Multisolvent contact angle measurements were used to calculate surface free energy and acid-base polar components of bare and modified surfaces based on the van Oss-Chaudhury-Good model: results indicate that protein resistance in these phenylglycoside layers correlates positively with wetting behavior and Lewis basicity. PMID:27108562

  2. A surface immobilization method of endoglucanase from Cellulomonas biazotea mutant improved catalytic properties of biocatalyst during processing.

    Science.gov (United States)

    Rajoka, Muhammad Ibrahim; Zia, Yasmin

    2007-01-01

    Purified endoglucanase from C. biazotea mutant 51SM(r) was successfully immobilized on Eudragit L-100, with 75 % yield of immobilization. This method improved the kinetic and thermodynamic properties of the enzyme. Immobilization significantly decreased entropy and enthalpy of inactivation of biocatalyst and made it functionally and thermodynamically more stable and reusable compared to free one.

  3. Research of the influencing factors of the immobilized redox mediators accelerating Cr(Ⅵ reduction by Escherichia coli BL21

    Directory of Open Access Journals (Sweden)

    Jiapeng PING

    2017-10-01

    Full Text Available In order to study the influencing factors of non-dissolved redox mediators on Cr (Ⅵ bioreduction, the accelerating effect of non-dissolved redox mediators immobilized by cellulose acetate (CA on the reduction of Cr(Ⅵ by Escherichia coli BL21 is investigated. The results show that the accelerating order is 1-chloroanthraquinone>1,5-dichloroanthraquinone>2-chloroanthraquinone>1,8-dichloroanthraquinone>anthraquinone>1,4,5,8-tetrachloroanthtaquinone; the optimal concentration of 1-chloroanthraquinone pH and initial Cr(Ⅵ concentration are 0.048 mol/L, 7.00 and 30 mg/L, respectively; the removal rate increases with the increasing temperature when it is in the range of 20~60 ℃; after 6 times of recycling experiments, and the Cr(Ⅵ bioreduction rate with the immobilized 1-chloroanthraquinone maintains above 5 times of margin bacterium's. It indicates that non-dissolved redox mediators immobilized by CA can effectively accelerate the reduction rate of Cr(Ⅵ by Escherichia coli BL21, which has favourable application value.

  4. Tailoring the surface properties of polypropylene films through cold atmospheric pressure plasma (CAPP) assisted polymerization and immobilization of biomolecules for enhancement of anti-coagulation activity

    Energy Technology Data Exchange (ETDEWEB)

    Navaneetha Pandiyaraj, K., E-mail: dr.knpr@gmail.com [Surface Engineering Laboratory, Department of Physics, Sri Shakthi Institute of Engineering and Technology, L& T By Pass, Chinniyam Palayam (Post), Coimbatore 641062 (India); Ram Kumar, M.C.; Arun Kumar, A. [Surface Engineering Laboratory, Department of Physics, Sri Shakthi Institute of Engineering and Technology, L& T By Pass, Chinniyam Palayam (Post), Coimbatore 641062 (India); Padmanabhan, P.V.A. [PSN College of Engineering and Technology, Tirunelveli 627 152 (India); Deshmukh, R.R. [Department of Physics, Institute of Chemical Technology, Matunga, Mumbai 400 019 (India); Bah, M.; Ismat Shah, S. [Department of Physics and Astronomy, Department of Materials Science and Engineering, University of Delaware, 208 Dupont Hall, Newark (United States); Su, Pi-Guey [Department of Chemistry, Chinese Culture University, Taipei 111, Taiwan (China); Halleluyah, M.; Halim, A.S. [School of Medical Sciences, Health Campus, Universiti Sains Malaysia, 16150 Kubang Kerian, Kelantan (Malaysia)

    2016-05-01

    Graphical abstract: - Highlights: • Developed low cost cold atmospheric plasma reactor for plasma polymerization technique. • Surface of the PP film was modified by grafting of AAc and PEG by CAPP polymerization. • Biomolecules of chitosan, insulin and heparin were immobilized on surface of PEG-AAc grafted PP films. • The surface modified PP films were characterized by various techniques. • The plasma polymerized and immobilized film reveals substantial blood compatibility. - Abstract: Enhancement of anti-thrombogenic properties of polypropylene (PP) to avert the adsorption of plasma proteins (fibrinogen and albumin), adhesion and activation of the platelets are very important for vast biomedical applications. The cold atmospheric pressure plasma (CAPP) assisted polymerization has potential to create the specific functional groups such as O−C=O, C=O, C−N and S−S. on the surface of polymeric films using selective precursor in vapour phase to enhance anti-thrombogenic properties. Such functionalized polymeric surfaces would be suitable for various biomedical applications especially to improve the blood compatibility. The eventual aspiration of the present investigation is to develop the biofunctional coating onto the surface of PP films using acrylic acid (AAc) and polyethylene glycol (PEG) as a precursor in a vapour phase by incorporating specific functional groups for immobilization of biomolecules such as heparin (HEP), chitosan (CHI) and insulin (INS) on the surface of plasma modified PP films. The surface properties such as hydrophilicity, chemical composition, surface topography of the surface modified PP films were analyzed by contact angle (CA), Fourier transform infrared spectroscopy (FTIR), X-ray photo electron spectroscopy (XPS) and atomic force microscopy (AFM). Furthermore the anti-thrombogenic properties of the surface modified PP films were studied by in vitro tests which include platelet adhesion and protein adsorption analysis. It was

  5. Tailoring the surface properties of polypropylene films through cold atmospheric pressure plasma (CAPP) assisted polymerization and immobilization of biomolecules for enhancement of anti-coagulation activity

    International Nuclear Information System (INIS)

    Navaneetha Pandiyaraj, K.; Ram Kumar, M.C.; Arun Kumar, A.; Padmanabhan, P.V.A.; Deshmukh, R.R.; Bah, M.; Ismat Shah, S.; Su, Pi-Guey; Halleluyah, M.; Halim, A.S.

    2016-01-01

    Graphical abstract: - Highlights: • Developed low cost cold atmospheric plasma reactor for plasma polymerization technique. • Surface of the PP film was modified by grafting of AAc and PEG by CAPP polymerization. • Biomolecules of chitosan, insulin and heparin were immobilized on surface of PEG-AAc grafted PP films. • The surface modified PP films were characterized by various techniques. • The plasma polymerized and immobilized film reveals substantial blood compatibility. - Abstract: Enhancement of anti-thrombogenic properties of polypropylene (PP) to avert the adsorption of plasma proteins (fibrinogen and albumin), adhesion and activation of the platelets are very important for vast biomedical applications. The cold atmospheric pressure plasma (CAPP) assisted polymerization has potential to create the specific functional groups such as O−C=O, C=O, C−N and S−S. on the surface of polymeric films using selective precursor in vapour phase to enhance anti-thrombogenic properties. Such functionalized polymeric surfaces would be suitable for various biomedical applications especially to improve the blood compatibility. The eventual aspiration of the present investigation is to develop the biofunctional coating onto the surface of PP films using acrylic acid (AAc) and polyethylene glycol (PEG) as a precursor in a vapour phase by incorporating specific functional groups for immobilization of biomolecules such as heparin (HEP), chitosan (CHI) and insulin (INS) on the surface of plasma modified PP films. The surface properties such as hydrophilicity, chemical composition, surface topography of the surface modified PP films were analyzed by contact angle (CA), Fourier transform infrared spectroscopy (FTIR), X-ray photo electron spectroscopy (XPS) and atomic force microscopy (AFM). Furthermore the anti-thrombogenic properties of the surface modified PP films were studied by in vitro tests which include platelet adhesion and protein adsorption analysis. It was

  6. Pinning of the Contact Line during Evaporation on Heterogeneous Surfaces: Slowdown or Temporary Immobilization? Insights from a Nanoscale Study.

    Science.gov (United States)

    Zhang, Jianguo; Müller-Plathe, Florian; Leroy, Frédéric

    2015-07-14

    The question of the effect of surface heterogeneities on the evaporation of liquid droplets from solid surfaces is addressed through nonequilibrium molecular dynamics simulations. The mechanism behind contact line pinning which is still unclear is discussed in detail on the nanoscale. Model systems with the Lennard-Jones interaction potential were employed to study the evaporation of nanometer-sized cylindrical droplets from a flat surface. The heterogeneity of the surface was modeled through alternating stripes of equal width but two chemical types. The first type leads to a contact angle of 67°, and the other leads to a contact angle of 115°. The stripe width was varied between 2 and 20 liquid-particle diameters. On the surface with the narrowest stripes, evaporation occurred at constant contact angle as if the surface was homogeneous, with a value of the contact angle as predicted by the regular Cassie-Baxter equation. When the width was increased, the contact angle oscillated during evaporation between two boundaries whose values depend on the stripe width. The evaporation behavior was thus found to be a direct signature of the typical size of the surface heterogeneity domains. The contact angle both at equilibrium and during evaporation could be predicted from a local Cassie-Baxter equation in which the surface composition within a distance of seven fluid-particle diameters around the contact line was considered, confirming the local nature of the interactions that drive the wetting behavior of droplets. More importantly, we propose a nanoscale explanation of pinning during evaporation. Pinning should be interpreted as a drastic slowdown of the contact line dynamics rather than a complete immobilization of it during a transition between two contact angle boundaries.

  7. Immobilization of heparin on the surface of polypropylene non-woven fabric for improvement of the hydrophilicity and blood compatibility.

    Science.gov (United States)

    Li, Rong; Wang, Hengdong; Wang, Wenfeng; Ye, Yin

    2013-01-01

    A polypropylene non-woven fabric (PPNWF) was exposed to oxygen plasma to produce peroxides on its surface. These peroxides were used to initiate graft polymerization of acrylic acid (AA) on the surface of PPNWF. Direct heparinization was accomplished via a reaction between heparin and PP-PAA (AA grafted PPNWF) which was activated by EDC (N-ethyl-N'-[3-(dimethylamino)propyl] carbodiimide). Indirect heparinized PPNWF was prepared by grafting poly(ethylene oxide) (PEO) on a PP-PAA surface to form PP-PAA-PEO, followed by reaction with heparin which was activated by EDC before use. The surface modified PPNWFs were characterized by attenuated total reflection Fourier transform infrared (ATR-FT-IR) spectroscopy, electron spectroscopy for chemical analysis (ESCA) and contact angle goniometry. It was found that hydrophilicity was greatly improved, as indicated by the decrease of the water contact angle from 142 to 33°. In vitro blood compatibility evaluation of modified PPNWFs, including hemolysis rate, platelet adhesion, plasma protein adsorption and activated partial thromboplastin time (APTT) was investigated. The results suggested that both heparinized PPNWFs showed lower hemolysis rates and better platelet anti-adhesion than non-heparinized controls. Furthermore, PPNWF obtained via indirect immobilization of heparin showed better hydrophilicity and blood compatibility than direct heparinization of PPNWF.

  8. Discrete complexes immobilized onto click-SBA-15 silica: controllable loadings and the impact of surface coverage on catalysis.

    Science.gov (United States)

    Nakazawa, Jun; Smith, Brian J; Stack, T Daniel P

    2012-02-08

    Azidopropyl functionalized mesoporous silica SBA-15 were prepared with variable azide loadings of 0.03-0.7 mmol g(-1) (~2-50% of maximal surface coverage) through a direct synthesis, co-condensation approach. These materials are functionalized selectively with ethynylated organic moieties through a copper-catalyzed azide alkyne cycloaddition (CuAAC) or "click" reaction. Specific loading within a material can be regulated by either the azide loading or limiting the alkyne reagent relative to the azide loading. The immobilization of ferrocene, pyrene, tris(pyridylmethyl)amine (TPA), and iron porphyrin (FeTPP) demonstrates the robust nature and reproducibility of this two-step synthetic attachment strategy. Loading-sensitive pyrene fluorescence correlates with a theoretically random surface distribution, rather than a uniform one; site-isolation of tethered moieties ~15 Å in length occurs at loadings less than 0.02 mmol g(-1). The effect of surface loading on reactivity is observed in the oxygenation of SBA-15-[Cu(I)(TPA)]. SBA-15-[Mn(II)(TPA)]-catalyzed epoxidation exhibits a systematic dependence on surface loading. A comparison of homogeneous, site-isolated and site-dense complexes provides insight into catalyst speciation and ligand activity.

  9. Porous Alpha-Tricalcium Phosphate with Immobilized Basic Fibroblast Growth Factor Enhances Bone Regeneration in a Canine Mandibular Bone Defect Model

    Directory of Open Access Journals (Sweden)

    Nobuhiro Kobayashi

    2016-10-01

    Full Text Available The effect of porous alpha-tricalcium phosphate (α-TCP with immobilized basic fibroblast growth factor (bFGF on bone regeneration was evaluated in a canine mandibular bone defect model. Identical bone defects were made in the canine mandible; six defects in each animal were filled with porous α-TCP with bFGF bound via heparin (bFGF group, whereas the other was filled with unmodified porous α-TCP (control group. Micro-computed tomography and histological evaluation were performed two, four and eight weeks after implantation. The bone mineral density of the bFGF group was higher than that of the control group at each time point (p < 0.05, and the bone mineral content of the bFGF group was higher than that of the control group at four and eight weeks (p < 0.05. Histological evaluation two weeks after implantation revealed that the porous α-TCP had degraded and bone had formed on the surface of α-TCP particles in the bFGF group. At eight weeks, continuous cortical bone with a Haversian structure covered the top of bone defects in the bFGF group. These findings demonstrate that porous α-TCP with immobilized bFGF can promote bone regeneration.

  10. Iron oxidation kinetics and phosphorus immobilization at the groundwater-surface water interface

    NARCIS (Netherlands)

    van der Grift, Bas|info:eu-repo/dai/nl/373433484; Rozemeijer, Joachim|info:eu-repo/dai/nl/304838403; Griffioen, Jasper|info:eu-repo/dai/nl/091129265; van der Velde, Ype

    2014-01-01

    Eutrophication of freshwater environments following diffuse nutrient loads is a widely recognized water quality problem in catchments. Fluxes of non-point P sources to surface waters originate from surface runoff and flow from soil water and groundwater into surface water. The availability of P in

  11. An inexpensive and simple method for thermally stable immobilization of DNA on an unmodified glass surface: UV linking of poly(T)10-poly(C)10-tagged DNA probes

    DEFF Research Database (Denmark)

    Guðnason, Haukur; Dufva, Hans Martin; Bang, Dang Duong

    2008-01-01

    the hybridization performance of the immobilized probes on the amino-silane surface, indicating a general benefit of adding a TC tag to DNA probes. In conclusion, our results show that using TC-tagged DNA probes immobilized on an unmodified glass surface is a robust, heat-stable, very simple, and inexpensive method...

  12. Immobilization of serum albumin and peptide aptamer for EPC on polydopamine coated titanium surface for enhanced in-situ self-endothelialization

    Energy Technology Data Exchange (ETDEWEB)

    Chen, Zhuoyue, E-mail: 362947953@qq.com [Key Lab. of Advanced Technology for Materials of Education Ministry, Southwest Jiaotong University, Chengdu, 610031 (China); RegeMed Lab of Tissue Engineering, Faculty of Life Science, Northwest University, Xi' an, 710069 (China); Li, Quanli [College of Stomology, Anhui Medical University, Hefei, 230032 (China); Chen, Jialong [Key Lab. of Advanced Technology for Materials of Education Ministry, Southwest Jiaotong University, Chengdu, 610031 (China); College of Stomology, Anhui Medical University, Hefei, 230032 (China); Luo, Rifang [Key Lab. of Advanced Technology for Materials of Education Ministry, Southwest Jiaotong University, Chengdu, 610031 (China); Maitz, Manfred F. [Key Lab. of Advanced Technology for Materials of Education Ministry, Southwest Jiaotong University, Chengdu, 610031 (China); Leibniz Institute of Polymer Research Dresden, Max Bergmann Center of Biomaterials, Dresden (Germany); Huang, Nan, E-mail: huangnan1956@163.com [Key Lab. of Advanced Technology for Materials of Education Ministry, Southwest Jiaotong University, Chengdu, 610031 (China)

    2016-03-01

    Restenosis and thrombosis are two major complications associated with vascular stents and grafts. The homing of circulating endothelial progenitor cells (EPCs) onto implant surfaces brings a new strategy to solve these problems by accelerating self -endothelialization in situ. Peptide aptamers with high affinity and specific recognition of EPCs can be immobilized to capture EPCs from the circulating blood. In this study, a biotinylated peptide aptamer (TPSLEQRTVYAK-GGGC-K-Biotin) for EPC, and bovine serum albumin (BSA) were co-immobilized onto titanium surface through avidin–biotin recognition to endow the surface with specific affinity for EPC and anti-platelet adhesion properties. Quartz crystal microbalance with dissipation (QCM-D), X-ray photoelectron spectroscopy (XPS), scanning electron microscopy (SEM) and water contact angle measuring were adopted for coating characterization. EPC affinity and hemocompatibility of the coating were also investigated in vitro. The results demonstrated that aptamer and BSA co-immobilized surface significantly reduced platelet adhesion and fibrinogen adsorption/activation. Besides, such functional surface could remarkably enhance EPC adhesion, without affecting the behavior of endothelial cells (ECs) and smooth muscle cells (SMCs) obviously. The result shows the possibility of utilizing such a multifunctional surface in cardiovascular implants. - Highlights: • We construct a multifunctional surface based on immobilization of BSA and aptamer. • It can significantly reduce platelet adhesion and fibrinogen adsorption/activation. • Such functional surface could remarkably enhance EPC adhesion in vitro. • It can induce rapid self-endothelialization of the implant surface in situ in vivo. • It is possible to use such a multifunctional surface in cardiovascular implants.

  13. Antibacterial and antibiofilm surfaces through Polydopamine-assisted immobilization of Lysostaphin as an antibacterial enzyme

    Science.gov (United States)

    Antibiotic resistance and the colonization of bacteria on surfaces, often as biofilms, prolong hospitalization periods, increase mortality, and are thus major concerns for health care providers. There is an urgent need for antimicrobial and antibiofilm surface treatments that are semi-permanent, can...

  14. Bio-functionalization of electro-synthesized polypyrrole surface by heme enzyme using a mixture of Nafion and glutaraldehyde as synergetic immobilization matrix: Conformational characterization and electrocatalytic studies

    Energy Technology Data Exchange (ETDEWEB)

    ElKaoutit, Mohammed, E-mail: elkaoutit@uca.es [Departamento de Quimica Analitica, Facultad de Ciencias, Universidad de Cadiz, 11510 Puerto Real, Cadiz (Spain); Naranjo-Rodriguez, Ignacio [Departamento de Quimica Analitica, Facultad de Ciencias, Universidad de Cadiz, 11510 Puerto Real, Cadiz (Spain); Dominguez, Manuel [Departamento de Fisica de la Materia Condensada, Facultad de Ciencias, Universidad de Cadiz, 11510 Puerto Real, Cadiz (Spain); Hidalgo-Hidalgo-de-Cisneros, Jose Luis [Departamento de Quimica Analitica, Facultad de Ciencias, Universidad de Cadiz, 11510 Puerto Real, Cadiz (Spain)

    2011-10-01

    Use of a mixture of Nafion and glutaraldehyde as new immobilization matrix was described. The percentage of Nafion was optimized to prevent denaturation of horseradish peroxidase enzyme after its crosslinkage with glutaraldehyde on electro-synthesized polypyrrole surface. Topographic study by Atomic Force Microscopy (AFM) shows that the enzyme seems to have been introduced inside the ionic cluster of Nafion. The characterization of the resulting bio-interfaces by UV-vis and FT-IR shows that the intra-crosslinkage phenomena caused by the use of glutaraldehyde can be eliminated by the optimization of the concentration of Nafion additive. The secondary structure contents of native and immobilized enzyme were analyzed by a Gaussian curve fitting of the respective FT-IR spectra in the amide I region. Immobilized enzyme presented notable increasing percentages of globular and short helical structure compared with native enzyme. This indicates that immobilized enzyme was folded which is in accordance with AFM studies and supports the enzyme entrance inside ionic clutter of Nafion. Thanks to synergic effects of the polypyrrole conducting polymer and the perfluorosulfonic acid polymer Nafion, HRP enzyme was immobilized in its 'native' state, the resulting biosensor was able to sense peroxide without any chemical mediator and can be categorized as third generation.

  15. Bio-functionalization of electro-synthesized polypyrrole surface by heme enzyme using a mixture of Nafion and glutaraldehyde as synergetic immobilization matrix: Conformational characterization and electrocatalytic studies

    International Nuclear Information System (INIS)

    ElKaoutit, Mohammed; Naranjo-Rodriguez, Ignacio; Dominguez, Manuel; Hidalgo-Hidalgo-de-Cisneros, Jose Luis

    2011-01-01

    Use of a mixture of Nafion and glutaraldehyde as new immobilization matrix was described. The percentage of Nafion was optimized to prevent denaturation of horseradish peroxidase enzyme after its crosslinkage with glutaraldehyde on electro-synthesized polypyrrole surface. Topographic study by Atomic Force Microscopy (AFM) shows that the enzyme seems to have been introduced inside the ionic cluster of Nafion. The characterization of the resulting bio-interfaces by UV-vis and FT-IR shows that the intra-crosslinkage phenomena caused by the use of glutaraldehyde can be eliminated by the optimization of the concentration of Nafion additive. The secondary structure contents of native and immobilized enzyme were analyzed by a Gaussian curve fitting of the respective FT-IR spectra in the amide I region. Immobilized enzyme presented notable increasing percentages of globular and short helical structure compared with native enzyme. This indicates that immobilized enzyme was folded which is in accordance with AFM studies and supports the enzyme entrance inside ionic clutter of Nafion. Thanks to synergic effects of the polypyrrole conducting polymer and the perfluorosulfonic acid polymer Nafion, HRP enzyme was immobilized in its 'native' state, the resulting biosensor was able to sense peroxide without any chemical mediator and can be categorized as third generation.

  16. Study of the surface crystallization and resistance to dissolution of niobium phosphate glasses for nuclear waste immobilization

    International Nuclear Information System (INIS)

    Vieira, Heveline

    2008-01-01

    The surface crystallization and the dissolution rate of three phosphate glass compositions containing different amounts of niobium oxide were studied. The glasses were named Nb30, Nb37, and Nb44 according to the nominal content of niobium oxide in the glass composition. The three compositions were evaluated keeping the P 2 O 5 /K 2 O ratio constant and varying the amount of Nb 2 O 5 . These glasses were produced by melting appropriate chemical compounds at 1500 deg C for 0.5 hour. The crystalline phases which were nucleated on the glass surface after heat treatment were determined by X-ray diffraction. The crystalline structures depend on the amount of niobium oxide in the glass composition. The crystal morphologies were observed by using an optical microscope, and their characteristics are specific for each kind of crystalline phase. The crystal growth rate and the surface nuclei density were determined for each glass composition, and they depend on each crystalline phase nucleated on the surface. From the differential thermal analysis curves it was determined that the Nb44 glass containing 46.5 mol por cent of niobium oxide is the most thermally stable against crystallization when compared to the Nb30 and Nb37 glasses. According to the activation energies determined for crystal growth on the surface of each glass type, the Nb44 glass can also be considered the most resistant one against crystallization. The dissolution rate for the Nb44 glass after 14 days immersed in an aqueous solution with pH equals to 7 at 90 deg C is the lowest (9.0 x 10 -7 g. cm -2 . day -1 ) when compared to the other two glass compositions. The dissolution rates in acidic and neutral solutions of all studied glasses meet the international standards for materials which can be used in the immobilization of nuclear wastes. (author)

  17. The Design of Simple Bacterial Microarrays: Development towards Immobilizing Single Living Bacteria on Predefined Micro-Sized Spots on Patterned Surfaces.

    Directory of Open Access Journals (Sweden)

    Nina Bjørk Arnfinnsdottir

    Full Text Available In this paper we demonstrate a procedure for preparing bacterial arrays that is fast, easy, and applicable in a standard molecular biology laboratory. Microcontact printing is used to deposit chemicals promoting bacterial adherence in predefined positions on glass surfaces coated with polymers known for their resistance to bacterial adhesion. Highly ordered arrays of immobilized bacteria were obtained using microcontact printed islands of polydopamine (PD on glass surfaces coated with the antiadhesive polymer polyethylene glycol (PEG. On such PEG-coated glass surfaces, bacteria were attached to 97 to 100% of the PD islands, 21 to 62% of which were occupied by a single bacterium. A viability test revealed that 99% of the bacteria were alive following immobilization onto patterned surfaces. Time series imaging of bacteria on such arrays revealed that the attached bacteria both divided and expressed green fluorescent protein, both of which indicates that this method of patterning of bacteria is a suitable method for single-cell analysis.

  18. Dynamic Factor Models for the Volatility Surface

    DEFF Research Database (Denmark)

    van der Wel, Michel; Ozturk, Sait R.; Dijk, Dick van

    The implied volatility surface is the collection of volatilities implied by option contracts for different strike prices and time-to-maturity. We study factor models to capture the dynamics of this three-dimensional implied volatility surface. Three model types are considered to examine desirable...

  19. Mass spectrometric investigation of synthetic glycoside of muramyl dipeptide immobilized on fumed silica surface

    International Nuclear Information System (INIS)

    Kulik, Tetiana V.; Azizova, Liana R.; Palyanytsya, Borys B.; Zemlyakov, Alexander E.; Tsikalova, Victoria N.

    2010-01-01

    N-Acetylmuramyl-L-alanyl-D-isoglutamine or muramyl dipeptide is a cleavage product of peptidoglycan by lysozyme. This study explored the use of the temperature-programmed desorption mass spectrometry (TPDMS) in analysis of glycoside of muramyl dipeptide: O-{(4-tert-butylcyclohexyl)-2-acetamido-2, 3-dideoxy-β-D-glucopyranoside-3-yl}-D-lactoyl-L-alanyl-D-isoglutamine (MDP) on the surface of fumed silica. Stages of pyrolysis of MDP in condensed state and on the silica surface have been determined. Three stages have been clear identified under pyrolysis of MDP on the silica surface. Kinetic parameters of thermal reactions on the fumed silica surface and in the condensed state have been calculated.

  20. Surface modification of polysulfone membranes applied for a membrane reactor with immobilized alcohol dehydrogenase

    DEFF Research Database (Denmark)

    Hoffmann, Christian; Silau, Harald; Pinelo, Manuel

    2018-01-01

    Commercially available polysulfone (PSf) membranes with a polypropylene backing are used across a broad range of applications. However, the natural properties of the PSf surface sometimes limit their application. Here we present, how the surface of supported membranes can be heterogeneously...... activated by lithiation followed by functionalization with acid chlorides at 0 °C, permitting modification of commercial PSf membranes without compromising the mechanical integrity of the membrane. Post-functionalization polymer grafting was illustrated through both, a “grafting from” approach by surface...... initiated atom transfer radical polymerization (SI-ATRP) and by a “grafting to” approach exploiting Cu(I) catalyzed 1,3-cycloadditions of alkynes with azides (CuAAC) introducing hydrophilic polymers onto the membrane surface. Poly(1-vinyl imidazole) (pVim) grafted membranes were exploited as support...

  1. Immobilized Rhizopus oryzae lipase catalyzed synthesis of palm stearin and cetyl alcohol wax esters: Optimization by Response Surface Methodology

    Directory of Open Access Journals (Sweden)

    Gargouri Youssef

    2011-06-01

    Full Text Available Abstract Background Waxes are esters of long-chain fatty acids and long-chain alcohols. Their principal natural sources are animals (sperm whale oil and vegetables (jojoba which are expensive and not easily available. Wax esters synthesized by enzymatic transesterification, using palm stearin as raw material, can be considered as an alternative to natural ones. Results Palm stearin is a solid fraction obtained by fractionation of palm oil. Palm stearin was esterified with cetyl alcohol to produce a mixture of wax esters. A non-commercial immobilized lipase from Rhizopus oryzae was used as biocatalyst. Response surface methodology was employed to determine the effects of the temperature (30-50°C, the enzyme concentration (33.34-300 IU/mL, the alcohol/palm stearin molar ratio (3-7 mol/mol and the substrate concentration (0.06-0.34 g/mL on the conversion yield of palm stearin. Under optimal conditions (temperature, 30°C; enzyme concentration, 300 IU/mL; molar ratio 3 and substrate concentration 0.21 g/mL a high conversion yield of 98.52% was reached within a reaction time of 2 h. Conclusions Response surface methodology was successfully applied to determine the optimum operational conditions for synthesis of palm stearin based wax esters. This study may provide useful tools to develop economical and efficient processes for the synthesis of wax esters.

  2. Immobilized Rhizopus oryzae lipase catalyzed synthesis of palm stearin and cetyl alcohol wax esters: optimization by response surface methodology.

    Science.gov (United States)

    Sellami, Mohamed; Aissa, Imen; Frikha, Fakher; Gargouri, Youssef; Miled, Nabil

    2011-06-17

    Waxes are esters of long-chain fatty acids and long-chain alcohols. Their principal natural sources are animals (sperm whale oil) and vegetables (jojoba) which are expensive and not easily available. Wax esters synthesized by enzymatic transesterification, using palm stearin as raw material, can be considered as an alternative to natural ones. Palm stearin is a solid fraction obtained by fractionation of palm oil. Palm stearin was esterified with cetyl alcohol to produce a mixture of wax esters. A non-commercial immobilized lipase from Rhizopus oryzae was used as biocatalyst. Response surface methodology was employed to determine the effects of the temperature (30-50 °C), the enzyme concentration (33.34-300 IU/mL), the alcohol/palm stearin molar ratio (3-7 mol/mol) and the substrate concentration (0.06-0.34 g/mL) on the conversion yield of palm stearin. Under optimal conditions (temperature, 30 °C; enzyme concentration, 300 IU/mL; molar ratio 3 and substrate concentration 0.21 g/mL) a high conversion yield of 98.52% was reached within a reaction time of 2 h. Response surface methodology was successfully applied to determine the optimum operational conditions for synthesis of palm stearin based wax esters. This study may provide useful tools to develop economical and efficient processes for the synthesis of wax esters. © 2011 Sellami et al; licensee BioMed Central Ltd.

  3. Immobilized Rhizopus oryzae lipase catalyzed synthesis of palm stearin and cetyl alcohol wax esters: Optimization by Response Surface Methodology

    Science.gov (United States)

    2011-01-01

    Background Waxes are esters of long-chain fatty acids and long-chain alcohols. Their principal natural sources are animals (sperm whale oil) and vegetables (jojoba) which are expensive and not easily available. Wax esters synthesized by enzymatic transesterification, using palm stearin as raw material, can be considered as an alternative to natural ones. Results Palm stearin is a solid fraction obtained by fractionation of palm oil. Palm stearin was esterified with cetyl alcohol to produce a mixture of wax esters. A non-commercial immobilized lipase from Rhizopus oryzae was used as biocatalyst. Response surface methodology was employed to determine the effects of the temperature (30-50°C), the enzyme concentration (33.34-300 IU/mL), the alcohol/palm stearin molar ratio (3-7 mol/mol) and the substrate concentration (0.06-0.34 g/mL) on the conversion yield of palm stearin. Under optimal conditions (temperature, 30°C; enzyme concentration, 300 IU/mL; molar ratio 3 and substrate concentration 0.21 g/mL) a high conversion yield of 98.52% was reached within a reaction time of 2 h. Conclusions Response surface methodology was successfully applied to determine the optimum operational conditions for synthesis of palm stearin based wax esters. This study may provide useful tools to develop economical and efficient processes for the synthesis of wax esters. PMID:21682865

  4. Mechanism of the immobilization of surfactants on polymeric surfaces by means of an argon plasma treatment: Influence of UV radiation

    NARCIS (Netherlands)

    Lens, J.P.; Spaay, B.; Terlingen, J.G.A.; Engbers, G.H.M.; Feijen, Jan

    1999-01-01

    The mechanism of the immobilization of the surfactant sodium 10-undecenoate (C11(:)) on poly(ethylene) (PE) by means of an argon plasma treatment has been investigated. In particular, the influence of the vacuum ultraviolet (UV) radiation emitted by the argon plasma on the immobilization was

  5. Optimization of the production of biodiesel by a commercial immobilized lipase in a solvent-free system using a response surface methodology

    Directory of Open Access Journals (Sweden)

    ZORICA KNEZEVIC

    2008-02-01

    Full Text Available Response surface methodology was used for the evaluation of the effects of various factors on the synthesis of biodiesel catalyzed with immobilized lipase from Rhizomucor miehei in a solvent-free system. The production of biodiesel was optimized and model response equations were obtained, enabling the prediction of biodiesel production from the values of the four main factors. It would seem that the reaction temperature and the amount of water predominantly determined the conversion process while the methanol/oil molar ratio had no significant influence on the reaction rate. The temperature and amount of water showed negative interactive effects on the observed reaction rate per amount of enzyme. However, there were no significant interactions among the other variables according to the test of statistical significance. The highest yield of 10.15 mol kg-1 enzyme was observed at 45 °C with a 6:1 methanol to oil molar ratio and with no added water in the system.

  6. Electrospun regenerated cellulose nanofibrous membranes surface-grafted with polymer chains/brushes via the atom transfer radical polymerization method for catalase immobilization.

    Science.gov (United States)

    Feng, Quan; Hou, Dayin; Zhao, Yong; Xu, Tao; Menkhaus, Todd J; Fong, Hao

    2014-12-10

    In this study, an electrospun regenerated cellulose (RC) nanofibrous membrane with fiber diameters of ∼200-400 nm was prepared first; subsequently, 2-hydroxyethyl methacrylate (HEMA), 2-dimethylaminoethyl methacrylate (DMAEMA), and acrylic acid (AA) were selected as the monomers for surface grafting of polymer chains/brushes via the atom transfer radical polymerization (ATRP) method. Thereafter, four nanofibrous membranes (i.e., RC, RC-poly(HEMA), RC-poly(DMAEMA), and RC-poly(AA)) were explored as innovative supports for immobilization of an enzyme of bovine liver catalase (CAT). The amount/capacity, activity, stability, and reusability of immobilized catalase were evaluated, and the kinetic parameters (Vmax and Km) for immobilized and free catalase were determined. The results indicated that the respective amounts/capacities of immobilized catalase on RC-poly(HEMA) and RC-poly(DMAEMA) nanofibrous membranes reached 78 ± 3.5 and 67 ± 2.7 mg g(-1), which were considerably higher than the previously reported values. Meanwhile, compared to that of free CAT (i.e., 18 days), the half-life periods of RC-CAT, RC-poly(HEMA)-CAT, RC-poly(DMAEMA)-CAT, and RC-poly(AA)-CAT were 49, 58, 56, and 60 days, respectively, indicating that the storage stability of immobilized catalase was also significantly improved. Furthermore, the immobilized catalase exhibited substantially higher resistance to temperature variation (tested from 5 to 70 °C) and lower degree of sensitivity to pH value (tested from 4.0 and 10.0) than the free catalase. In particular, according to the kinetic parameters of Vmax and Km, the nanofibrous membranes of RC-poly(HEMA) (i.e., 5102 μmol mg(-1) min(-1) and 44.89 mM) and RC-poly(DMAEMA) (i.e., 4651 μmol mg(-1) min(-1) and 46.98 mM) had the most satisfactory biocompatibility with immobilized catalase. It was therefore concluded that the electrospun RC nanofibrous membranes surface-grafted with 3-dimensional nanolayers of polymer chains/brushes would be

  7. Surface-immobilized hydrogel patterns on length scales from micrometer to nanometer

    Science.gov (United States)

    Zeira, Assaf

    The present work concentrates on the study of pattern generation and transfer processes of monolayer covered surfaces, deriving from the basic working concept of Constructive Lithography. As an advancement of constructive lithography, we developed a direct, one-step printing (contact electrochemical printing, CEP) and replication (contact electrochemical replication, CER) of hydrophilic organic monolayer patterns surrounded by a hydrophobic monolayer background. In addition, we present a process of transfer of metal between two contacting solid surfaces to predefined monolayer template pattern sites (contact electrochemical transfer, CET). This thesis shows that CEP, CER, and CET may be implemented under a variety of different experimental conditions, regardless of whether the initial "master" pattern was created by a parallel (fast) or serial (slow) patterning process. CEP and CER also posses the unique attractive property that each replica may equally function as master stamp in the fabrication of additional replicas. Moreover, due to a mechanism of selfcorrection patterned surfaces produced these process are often free of defects that the initial "master" stamp may had. We finally show that the electrochemical patterning of OTS monolayers on silicon can be further extended to flexible polymeric substrate materials as well as to a variety of chemical manipulations, allowing the fabrication of tridimensional (3D) composite structures made on the basis of readily available OTS compound. The results obtained suggest that such contact electrochemical processes could be used to rapidly generate multiple copies of surface patterns spanning variable length scales, this basic approach being applicable to rigid as well as flexible substrate materials.

  8. Tissue factor-expressing tumor cells can bind to immobilized recombinant tissue factor pathway inhibitor under static and shear conditions in vitro.

    Directory of Open Access Journals (Sweden)

    Sara P Y Che

    Full Text Available Mammary tumors and malignant breast cancer cell lines over-express the coagulation factor, tissue factor (TF. High expression of TF is associated with a poor prognosis in breast cancer. Tissue factor pathway inhibitor (TFPI, the endogenous inhibitor of TF, is constitutively expressed on the endothelium. We hypothesized that TF-expressing tumor cells can bind to immobilized recombinant TFPI, leading to arrest of the tumor cells under shear in vitro. We evaluated the adhesion of breast cancer cells to immobilized TFPI under static and shear conditions (0.35 - 1.3 dyn/cm2. We found that high-TF-expressing breast cancer cells, MDA-MB-231 (with a TF density of 460,000/cell, but not low TF-expressing MCF-7 (with a TF density of 1,400/cell, adhered to recombinant TFPI, under static and shear conditions. Adhesion of MDA-MB-231 cells to TFPI required activated factor VII (FVIIa, but not FX, and was inhibited by a factor VIIa-blocking anti-TF antibody. Under shear, adhesion to TFPI was dependent on the TFPI-coating concentration, FVIIa concentration and shear stress, with no observed adhesion at shear stresses greater than 1.0 dyn/cm2. This is the first study showing that TF-expressing tumor cells can be captured by immobilized TFPI, a ligand constitutively expressed on the endothelium, under low shear in vitro. Based on our results, we hypothesize that TFPI could be a novel ligand mediating the arrest of TF-expressing tumor cells in high TFPI-expressing vessels under conditions of low shear during metastasis.

  9. Novel procedure to enhance PLA surface properties by chitosan irreversible immobilization

    Energy Technology Data Exchange (ETDEWEB)

    Stoleru, Elena; Dumitriu, Raluca Petronela [Petru Poni Institute of Macromolecular Chemistry, Grigore Ghica Voda Alley, 41A, 700487 Iasi (Romania); Munteanu, Bogdanel Silvestru [“Al. I. Cuza” University, Faculty of Physics, 11 Carol I Blvd., 700506 Iasi (Romania); Zaharescu, Traian [INCDIE ICPE CA, Bucharest (Romania); Tănase, Elisabeta Elena; Mitelut, Amalia [Industrial Biotechnology Department, Faculty of Biotechnology – USAMV Bucharest (Romania); Ailiesei, Gabriela-Liliana [Petru Poni Institute of Macromolecular Chemistry, Grigore Ghica Voda Alley, 41A, 700487 Iasi (Romania); Vasile, Cornelia, E-mail: cvasile@icmpp.ro [Petru Poni Institute of Macromolecular Chemistry, Grigore Ghica Voda Alley, 41A, 700487 Iasi (Romania)

    2016-03-30

    Graphical abstract: - Highlights: • PLA requires functionalization prior to surface attaching chitosan. • Chitosan with different molecular weights was grafted onto PLA surface. • Antibacterial, antifungal, antioxidant PLA-based materials are obtained. • Nano-fibers coatings obtained by electrospinning of high molecular weight chitosan. - Abstract: A novel two step procedure was applied for poly(lactic acid) (PLA) functionalization consisting in the exposure to cold radiofrequency plasma in nitrogen atmosphere or to gamma irradiation followed by “grafting to” of a chitosan layer using carbodiimide chemistry. The adhesion and stability of the deposited surface layer was assured by plasma/gamma irradiation treatment while the chitosan layer offers antifungal/antibacterial/antioxidant activities. Chitosan with different viscosities/deacetylation degree was deposited by electrospinning or immersion methods. Correlations between rheological behavior of chitosan solutions and chitosan layer deposition conditions are made. The PLA surface properties were investigated by water contact angle measurements, ATR-FTIR spectroscopy, AFM, chemiluminiscence, etc. It has been established that the surface roughness increases direct proportional with cold plasma duration and gamma irradiation dose and further increases by chitosan coating which at its turn depends on chitosan characteristics (viscosity and deacetylation degree) and method of deposition. Nano-fibers with relatively homogeneous and reproducible features are obtained by electrospinning of highly viscous chitosan while with the other two types of chitosan both microparticles and nano-fibers are formed. The chitosan coating obtained by immersion is more homogenous and compact and has a better antibacterial activity than the electrospun layer as fiber meshes.

  10. The Effect of Surface Functionalization on the Immobilization of Gold Nanoparticles on Graphene Sheets

    OpenAIRE

    Song, Min; Xu, Juan; Wu, Changzi

    2012-01-01

    In our study, graphene oxide is synthesized by Hummers method. And then, carboxylic acid functionalized graphene (graphene-COOH), thiol-functionalized graphene (graphene-SH), and highly dispersive graphene are prepared by chemical modification of respective groups on the graphene surface. Furthermore, we explore a solution-based approach to prepare three differently functionalized graphene-gold composites by one-step chemical reduction of AuCl4 - ions in respective functionalized graphene sus...

  11. Fabricating a reactive surface on the fibroin film by a room-temperature plasma jet array for biomolecule immobilization

    Science.gov (United States)

    Chen, Guang-Liang; Zheng, Xu; Lü, Guo-Hua; Zhang, Zhao-Xia; Sylvain, Massey; Wilson, Smith; Michael, Tatoulian; Yang, Si-Ze

    2012-10-01

    A simple dielectric barrier discharge (DBD) jet array was designed with a liquid electrode and helium gas. The characteristics of the jet array discharge and the preliminary polymerization with acrylic acid (AA) monomer were presented. The plasma reactor can produce a cold jet array with a gas temperature lower than 315 K, using an applied discharge power between 6 W and 30 W (Vdis × Idis). A silk fibroin film (SFF) was modified using the jet array and AA monomer, and the treated SFF samples were characterized by atomic force microscopy (AFM), scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR), and contact angle (CA). The deposition rate of the poly acrylic acid (PAA) was able to reach 300 nm/min, and the surface roughness and energy increased with the AA flow rate. The FTIR results indicate that the modified SFF had more carboxyl groups (-COOH) than the original SFF. This latter characteristic allowed the modified SFF to immobilize more quantities of antimicrobial peptide (AP, LL-37) which inhibited the Escherichia coli (E. Coli) effectively.

  12. Photochemical cleaving of allochthonous organic-metal complexes contributes to phosphorus immobilization in surface waters

    Czech Academy of Sciences Publication Activity Database

    Porcal, Petr; Frejlachová, K.; Kopáček, Jiří; Nedoma, Jiří; Šavrdová, T.

    2017-01-01

    Roč. 167, JAN (2017), s. 374-381 ISSN 0045-6535 R&D Projects: GA ČR GA15-09721S Institutional support: RVO:60077344 Keywords : photodegradation * dom * phosphorus * aluminum * iron Subject RIV: DA - Hydrology ; Limnology OBOR OECD: Environmental sciences (social aspects to be 5.7) Impact factor: 4.208, year: 2016

  13. Immobilization of enzymes using non-ionic colloidal liquid aphrons (CLAs): Surface and enzyme effects.

    Science.gov (United States)

    Ward, Keeran; Xi, Jingshu; Stuckey, David C

    2015-12-01

    The use of non-ionic colloidal liquid aphrons (CLAs) as a support for enzyme immobilisation was investigated. Formulation required the mixing of an aqueous-surfactant solution with a relatively non-polar solvent-surfactant solution, forming a solvent droplet surrounded by a thin stabilised aqueous film (soapy shell). Studies utilising anionic surfactants have showed increased retention, however, very little have been understood about the forces governing immobilisation. This study seeks to determine the effects of enzyme properties on CLA immobilisation by examining a non-ionic/non-polar solvent system comprised of two non-ionic surfactants, Tween 20 and 80, mineral oil and the enzymes lipase, aprotinin and α-chymotrypsin. From these results it was deduced that hydrophobic interactions strongly governed immobilisation. Confocal Scanning Laser Microscopy (CSLM) revealed that immobilisation was predominantly achieved by surface adsorption attributed to hydrophobic interactions between the enzyme and the CLA surface. Enzyme surface affinity was found to increase when added directly to the formulation (pre-manufacture addition), as opposed to the bulk continuous phase (post-manufacture addition), with α-chymotrypsin and aprotinin being the most perturbed, while lipase was relatively unaffected. The effect of zeta potential on immobilisation showed that enzymes adsorbed better closer to their pI, indicating that charge minimisation was necessary for immobilisation. Finally, the effect of increasing enzyme concentration in the aqueous phase resulted in an increase in adsorption for all enzymes due to cooperativity between protein molecules, with saturation occurring faster at higher adsorption rates. Copyright © 2015 Elsevier B.V. All rights reserved.

  14. Immobilization of poly(MPC) brushes onto titanium surface by combining dopamine self-polymerization and ATRP: Preparation, characterization and evaluation of hemocompatibility in vitro

    Energy Technology Data Exchange (ETDEWEB)

    Ma, Wenyong; Yang, Ping; Li, Jingan; Li, Shiqi; Li, Peichuang; Zhao, Yuancong, E-mail: Zhaoyc7320@163.com; Huang, Nan

    2015-09-15

    Graphical abstract: The functional surface containing rich amino and hydroxyl groups was obtained by simple and easily dopamine self-polymerization. Poly (MPC) brushes were successfully immobilized on titanium surface by combining acylation reaction and ATRP. This chemical and biomimetic modified titanium surface effectively inhibits platelet adhesion and activation. - Highlights: • Polydopamine coating provides amino and hydroxyl groups for second reactivity. • Poly(MPC) brushes covalently immobilized on coating by surface initiated ATRP. • In vitro hemocompatibility of biomimetic modified Ti was better than unmodified. - Abstract: Poly(2-methacryloyloxyethyl phosphorylcholine(MPC)) has been studied in many biomedical fields because of good biocompatibility, such as hemocompatibility, inhibiting protein adhesion, antifouling, and so on. To achieve good hemocompatibility of titanium (Ti) surface, bio-inspired poly(MPC) brushes were grafted from Ti substrate covalently. In this work, the surface of Ti was first coated with polydopamine, and got a surface bearing −NH{sub 2} and −OH groups which could be bridged with poly(MPC) via atom transfer radical polymerization. Water contact angle decreased to 51.3° when titanium grafted with poly(MPC) brushes. The data of Infrared Spectroscopy and X-ray photoelectron spectroscopy (XPS) indicated that poly(MPC) was successfully grafted onto the surface of titanium. Platelet-rich plasma (PRP) adhesion test and lactate dehydrogenase (LDH) assay showed that the number of platelets adhered on the surface of modified-titanium was much less than that of unmodified titanium and platelets did not aggregate and distort. Thus, the simple and chemical method of immobilization of poly(MPC) brushes has potential application for improving hemocompatibility for cardiovascular stent and some other biomaterials.

  15. In situ immobilized lipase on the surface of intracellular polyhydroxybutyrate granules: preparation, characterization, and its promising use for the synthesis of fatty acid alkyl esters.

    Science.gov (United States)

    Yang, Taek Ho; Kwon, Min-A; Lee, Ji Young; Choi, Ji-Eun; Oh, Joon Young; Song, Jae Kwang

    2015-12-01

    Photobacterium lipolyticum M37 lipase (LipM37) was immobilized on the surface of intracellular polyhydroxybutyrate (PHB) granules in Escherichia coli. LipM37 was genetically fused to Cupriavidus necator PHA synthase (PhaC Cn ), and the engineered PHB operon containing the lip M37 -phaC Cn successfully mediated the accumulation of PHB granules (85 wt.%) inside E. coli cells. The PHB granules were isolated from the crude cell extract, and the immobilized LipM37 was comparable with the free form of LipM37 except for a favorable increase in thermostability. The immobilized LipM37 was used to synthesize oleic acid methyl ester (biodiesel) and oleic acid dodecyl ester (wax ester), and yielded 98.0 % conversion in esterification of oleic acid and dodecanol. It was suggested that the LipM37-PhaCCn fusion protein successfully exhibited bifunctional activities in E. coli and that in situ immobilization of lipase to the intracellular PHB could be a promising approach for expanding the biocatalytic toolbox for industrial chemical synthesis.

  16. Noncovalent immobilization of Pectinesterase (Prunus armeniaca L.) onto bentonite.

    Science.gov (United States)

    Karakuş, Emine; Ozler, Aynur; Pekyardimci, Sule

    2008-01-01

    In this work, pectinesterase isolated from Malatya apricot was immobilized onto acid-treated bentonite surface by simple adsorption at pH 9.0. The properties of free and immobilized enzyme were defined. The effect of various factors such as pH, temperature, heat, and storage stability on immobilized enzyme were investigated. Optimum pH and temperature were determined to be 9.0 and 50 degrees C, respectively. Kinetic parameters of the immobilized enzyme (Km and Vmax values) were also determined as 0.51 mM of the Km and 14.6 micromol min(-1) mg(-1) of the Vmax. No drastic change was observed in the Km value after immobilization. The Vmax value of immobilized enzyme was 8.4-fold bigger than those of free enzyme. Thermal and storage stability experiments were carried out. The patterns of heat stability indicated that the immobilization process tends to stabilize the enzyme. The properties of the immobilized enzyme were compared to those of the free enzyme.

  17. Immobilization of poly(acrylamide) brushes onto poly(caprolactone) surface by combining ATRP and “click” chemistry: Synthesis, characterization and evaluation of protein adhesion

    Energy Technology Data Exchange (ETDEWEB)

    Ma, Yuhao; Bian, Xinxiu; He, Liu; Cai, Mengtan; Xie, Xiaoxiong [College of Polymer Science and Engineering, Sichuan University, Chengdu 610065 (China); Luo, Xianglin, E-mail: luoxl@scu.edu.cn [College of Polymer Science and Engineering, Sichuan University, Chengdu 610065 (China); State Key Laboratory of Polymer Material and Engineering, Sichuan University, Chengdu 610065 (China)

    2015-02-28

    Highlights: • Poly(caprolacone) (PCL) film surface was chemically modified by a novel method through combining ATRP and “click” chemistry. • Poly(acrylamide) (PAAm) of tailored chain length were synthesized and “clicked” onto PCL surface. • The modified PCL surface showed reduced BSA and Fg adsorption, and the protein resist ability in terms of chain length through its impact on grafting reaction and modified surface was investigated. - Abstract: Developments of poly(caprolactone) in blood-contacting applications are often restricted due to its intrinsic hydrophobicity. One common way to improve its hemocompatibility is to attach hydrophilic polymers. Here we developed a non-destructive method to graft hydrophilic poly(acrylamide) (PAAm) onto poly(caprolactone) (PCL) surface. In this strategy, azido-ended PCL with low molecular weights was synthesized and blended with PCL to create a surface with “clickable” property. Alkyne-ended poly(acrylamide)s with controlled chain lengths were then synthesized by atom transfer radical polymerization (ATRP), and finally were immobilized onto PCL surface by “click” reaction. The occurrence of immobilization was verified qualitatively by water contact angle measurement and quantitatively by X-ray photoelectron spectroscopy (XPS). The PAAm grafted surface exhibited fouling resistant properties, as demonstrated by reduced bovine serum albumin (BSA) and fibrinogen (Fg) adhesion.

  18. Tailoring the surface properties of polypropylene films through cold atmospheric pressure plasma (CAPP) assisted polymerization and immobilization of biomolecules for enhancement of anti-coagulation activity

    Science.gov (United States)

    Navaneetha Pandiyaraj, K.; Ram Kumar, M. C.; Arun Kumar, A.; Padmanabhan, P. V. A.; Deshmukh, R. R.; Bah, M.; Ismat Shah, S.; Su, Pi-Guey; Halleluyah, M.; Halim, A. S.

    2016-05-01

    Enhancement of anti-thrombogenic properties of polypropylene (PP) to avert the adsorption of plasma proteins (fibrinogen and albumin), adhesion and activation of the platelets are very important for vast biomedical applications. The cold atmospheric pressure plasma (CAPP) assisted polymerization has potential to create the specific functional groups such as Osbnd Cdbnd O, Cdbnd O, Csbnd N and Ssbnd S. on the surface of polymeric films using selective precursor in vapour phase to enhance anti-thrombogenic properties. Such functionalized polymeric surfaces would be suitable for various biomedical applications especially to improve the blood compatibility. The eventual aspiration of the present investigation is to develop the biofunctional coating onto the surface of PP films using acrylic acid (AAc) and polyethylene glycol (PEG) as a precursor in a vapour phase by incorporating specific functional groups for immobilization of biomolecules such as heparin (HEP), chitosan (CHI) and insulin (INS) on the surface of plasma modified PP films. The surface properties such as hydrophilicity, chemical composition, surface topography of the surface modified PP films were analyzed by contact angle (CA), Fourier transform infrared spectroscopy (FTIR), X-ray photo electron spectroscopy (XPS) and atomic force microscopy (AFM). Furthermore the anti-thrombogenic properties of the surface modified PP films were studied by in vitro tests which include platelet adhesion and protein adsorption analysis. It was found that the anti-thrombogenic properties of the PP films are effectively controlled by the CAPP grafting of AAc and PEG followed by immobilization of biomolecules of heparin, chitosan and insulin. The grafting and immobilization was confirmed by FTIR and XPS through the recognition of specific functional groups such as COOH, Csbnd O, Ssbnd S and Csbnd N. on the surface of PP film. Furthermore, the surface morphology and hydrophilic nature of the PP films also tailored

  19. Surface functionalization of superparamagnetic nanoparticles encapsulated by chitosan for protein immobilization; Funcionalizacao da superficie de nanoparticulas superparamagneticas encapsuladas por quitosana para a imobilizacao de proteinas

    Energy Technology Data Exchange (ETDEWEB)

    Sousa, Jose Silva de

    2010-07-01

    Nanoscience and nanotechnology have opened up numerous developments of devices and systems on the nanometer scale, with new molecular organization, properties and functions. In this context, the polymeric magnetic nanoparticles are composites formed by magnetic materials with a particle size between 1 and 100 nm combined with functional polymers. They are well-known and have been widely studied because of its applications in various technology areas. Applications on the biological and medical areas include separation and immobilization of enzymes and proteins, improved techniques of magnetic resonance imaging and diagnostic systems for controlled drug delivery. In this work, proteins were immobilized on the surface of a biopolymer combined with superparamagnetic particles of magnetite. The biopolymer chitosan was used, cross-linked and functionalized with glutaraldehyde, applicable to the biological assays. Three types of magnetic composites were obtained, which were called QM1Glu, QM2NaGlu and QM3Glu. They were characterized by X-ray diffraction, scanning electron microscopy, vibrating sample magnetometry, differential scanning calorimetry, thermogravimetry and infrared spectroscopy. They were evaluated concerning the immobilization of the proteins bovine serum albumin (BSA), collagen and trypsin. The study showed that the immobilization of proteins on the biopolymer occurred in 30 min of incubation. The magnetic composite of non functionalized chitosan (QM3) was also evaluated. For trypsin, it was found that the immobilization potential of QM3 was higher than that observed for QM3Glu. After 30 days, the trypsin of the QM3-Trip and QM3Glu-Trip was still with activity. The activity and the enzyme kinetics of the QM3Glu-Trip with the substrate BApNA were demonstrated. (author)

  20. Novel approach for a PTX/VEGF dual drug delivery system in cardiovascular applications-an innovative bulk and surface drug immobilization.

    Science.gov (United States)

    Wulf, Katharina; Teske, Michael; Matschegewski, Claudia; Arbeiter, Daniela; Bajer, Dalibor; Eickner, Thomas; Schmitz, Klaus-Peter; Grabow, Niels

    2018-03-12

    The successive incorporation of several drugs into the polymeric bulk of implants mostly results in loss of considerable quantity of one drug, and/or the loss in quality of the coating and also in changes of drug release time points. A dual drug delivery system (DDDS) based on poly-L-lactide (PLLA) copolymers combining the effective inhibition of smooth muscle cell proliferation while simultaneously promoting re-endothelialization was successfully developed. To overcome possible antagonistic drug interactions and the limitation of the polymeric bulk material as release system for dual drugs, a novel concept which combines the bulk and surface drug immobilization for a DDDS was investigated. The advantage of this DDDS is that the bulk incorporation of fluorescein diacetate (FDAc) (model drug for paclitaxel (PTX)) via spray coating enhanced the subsequent cleavable surface coupling of vascular endothelial growth factor (VEGF) via the crosslinker bissulfosuccinimidyl suberate (BS 3 ). In the presence of the embedded FDAc, the VEGF loading and release are about twice times higher than in absence. Furthermore, the DDDS combines the diffusion drug delivery (FDAc or PTX) and the chemical controlled drug release, VEGF via hydrolysable ester bonds, without loss in quantity and quality of the drug release curves. Additionally, the performed in vitro biocompatibility study showed the bimodal influences of PTX and VEGF on human endothelial EA.hy926 cells. In conclusion, it was possible to show the feasibility to develop a novel DDDS which has a high potential for the medical application due to the possible easy and short modification of a polymer-based PTX delivery system.

  1. Decreased expression of extracellular matrix proteins and trophic factors in the amygdala complex of depressed mice after chronic immobilization stress

    Directory of Open Access Journals (Sweden)

    Jung Soonwoong

    2012-06-01

    Full Text Available Abstract Background The amygdala plays an essential role in controlling emotional behaviors and has numerous connections to other brain regions. The functional role of the amygdala has been highlighted by various studies of stress-induced behavioral changes. Here we investigated gene expression changes in the amygdala in the chronic immobilization stress (CIS-induced depression model. Results Eight genes were decreased in the amygdala of CIS mice, including genes for neurotrophic factors and extracellular matrix proteins. Among these, osteoglycin, fibromodulin, insulin-like growth factor 2 (Igf2, and insulin-like growth factor binding protein 2 (Igfbp2 were further analyzed for histological expression changes. The expression of osteoglycin and fibromodulin simultaneously decreased in the medial, basolateral, and central amygdala regions. However, Igf2 and Igfbp2 decreased specifically in the central nucleus of the amygdala. Interestingly, this decrease was found only in the amygdala of mice showing higher immobility, but not in mice displaying lower immobility, although the CIS regimen was the same for both groups. Conclusions These results suggest that the responsiveness of the amygdala may play a role in the sensitivity of CIS-induced behavioral changes in mice.

  2. Bioactive support for cell cultivation and potential grafting. Part1. Surface modification of 2-hydroxyethyl methacrylate hydrogels for avidin immobilization

    Czech Academy of Sciences Publication Activity Database

    Hobzová, Radka; Přádný, Martin; Zhunusbekova, Nazym M.; Širc, Jakub; Guryča, Vilém; Michálek, Jiří

    -, 043 (2011), s. 1-17 ISSN 1618-7229 R&D Projects: GA AV ČR 1QS400500558 Institutional research plan: CEZ:AV0Z40500505 Keywords : hydrogel * poly(2-hydroxyethyl methacrylate) * surface modification Subject RIV: CD - Macromolecular Chemistry Impact factor: 0.515, year: 2011 http://www.e-polymers.org/journal/abstract.cfm?abstract_Id=3616

  3. Probing the orientation of surface-immobilized protein G B1 using ToF-SIMS, sum frequency generation, and NEXAFS spectroscopy.

    Science.gov (United States)

    Baugh, Loren; Weidner, Tobias; Baio, J E; Nguyen, Phuong-Cac T; Gamble, Lara J; Stayton, Patrick S; Castner, David G

    2010-11-02

    The ability to orient active proteins on surfaces is a critical aspect of many medical technologies. An important related challenge is characterizing protein orientation in these surface films. This study uses a combination of time-of-flight secondary ion mass spectrometry (ToF-SIMS), sum frequency generation (SFG) vibrational spectroscopy, and near-edge X-ray absorption fine structure (NEXAFS) spectroscopy to characterize the orientation of surface-immobilized Protein G B1, a rigid 6 kDa domain that binds the Fc fragment of IgG. Two Protein G B1 variants with a single cysteine introduced at either end were immobilized via the cysteine thiol onto maleimide-oligo(ethylene glycol)-functionalized gold and bare gold substrates. X-ray photoelectron spectroscopy was used to measure the amount of immobilized protein, and ToF-SIMS was used to measure the amino acid composition of the exposed surface of the protein films and to confirm covalent attachment of protein thiol to the substrate maleimide groups. SFG and NEXAFS were used to characterize the ordering and orientation of peptide or side chain bonds. On both substrates and for both cysteine positions, ToF-SIMS data showed enrichment of mass peaks from amino acids located at the end of the protein opposite to the cysteine surface position as compared with nonspecifically immobilized protein, indicating end-on protein orientations. Orientation on the maleimide substrate was enhanced by increasing pH (7.0-9.5) and salt concentration (0-1.5 M NaCl). SFG spectral peaks characteristic of ordered α-helix and β-sheet elements were observed for both variants but not for cysteine-free wild type protein on the maleimide surface. The phase of the α-helix and β-sheet peaks indicated a predominantly upright orientation for both variants, consistent with an end-on protein binding configuration. Polarization dependence of the NEXAFS signal from the N 1s to π* transition of β-sheet peptide bonds also indicated protein ordering

  4. Probing the Orientation of Surface Immobilized Protein G B1 using ToF SIMS, Sum Frequency Generation, and NEXAFS Spectroscopy

    Science.gov (United States)

    Baugh, Loren; Weidner, Tobias; Baio, J.E.; Nguyen, Phuong Cac; Gamble, Lara J.; Stayton, Patrick S.; Castner, David G.

    2010-01-01

    The ability to orient active proteins on surfaces is a critical aspect of many medical technologies. An important related challenge is characterizing protein orientation in these surface films. This study uses a combination of time-of-flight secondary ion mass spectrometry (ToF-SIMS), sum frequency generation (SFG) vibrational spectroscopy, and near edge x-ray absorption fine structure (NEXAFS) spectroscopy to characterize the orientation of surface-immobilized Protein G B1, a rigid 6 kDa domain that binds the Fc fragment of IgG. Two Protein G B1 variants with a single cysteine introduced at either end were immobilized via the cysteine thiol onto maleimide-oligo(ethylene glycol)-functionalized gold and bare gold substrates. X-ray photoelectron spectroscopy was used to measure the amount of immobilized protein and ToF-SIMS was used to measure the amino acid composition of the exposed surface of the protein films and to confirm covalent attachment of protein thiol to the substrate maleimide groups. SFG and NEXAFS were used to characterize the ordering and orientation of peptide or side chain bonds. On both substrates and for both cysteine positions, ToF-SIMS data showed enrichment of mass peaks from amino acids located at the end of the protein opposite the cysteine surface position compared with nonspecifically immobilized protein, indicating end-on protein orientations. Orientation on the maleimide substrate was enhanced by increasing pH (7.0 to 9.5) and salt concentration (0 to 1.5 M NaCl). SFG spectral peaks characteristic of ordered α-helix and β-sheet elements were observed for both variants but not for cysteine-free wild type protein on the maleimide surface. The phase of the α-helix and β-sheet peaks indicated a predominantly upright orientation for both variants, consistent with an end-on protein binding configuration. Polarization dependence of the NEXAFS signal from the N 1s toπ* transition of β-sheet peptide bonds also indicated protein ordering

  5. Microorganism immobilization

    Science.gov (United States)

    Compere, Alicia L.; Griffith, William L.

    1981-01-01

    Live metabolically active microorganisms are immobilized on a solid support by contacting particles of aggregate material with a water dispersible polyelectrolyte such as gelatin, crosslinking the polyelectrolyte by reacting it with a crosslinking agent such as glutaraldehyde to provide a crosslinked coating on the particles of aggregate material, contacting the coated particles with live microorganisms and incubating the microorganisms in contact with the crosslinked coating to provide a coating of metabolically active microorganisms. The immobilized microorganisms have continued growth and reproduction functions.

  6. Immobilization of surface active compounds on polymer supports using glow discharge processess. 1. Sodium dodecyl sulfate on poly(propylene)

    NARCIS (Netherlands)

    Terlingen, J.G.A.; Terlingen, Johannes G.A.; Feijen, Jan; Hoffman, Allan S.

    1993-01-01

    A new method has been developed in which a reversibly adsorbed layer of a surfactant (sodium dodecyl sulfate, SDS) is covalently immobilized in one step onto a hydrophobic substrate (poly(propylene), PP) by applying an argon plasma treatment. The adsorption of SDS from aqueous solutions onto PP

  7. Comparison of artificial neural network (ANN) and response surface methodology (RSM) in optimization of the immobilization conditions for lipase from Candida rugosa on Amberjet(®) 4200-Cl.

    Science.gov (United States)

    Fatiha, Benamia; Sameh, Bouchagra; Youcef, Saihi; Zeineddine, Djeghaba; Nacer, Rebbani

    2013-01-01

    Candida rugosa lipase (CRL) is an important industrial enzyme that is successfully utilized in a variety of hydrolysis and esterification reactions. This work describes the optimization of immobilization conditions (enzyme/support ratio, immobilization temperature, and buffer concentration) of CRL on the anionic resin Amberjet® 4200-Cl, using enantioselectivity (E) as the reference parameter. The model reaction used for this purpose is the acylation of (R,S)-1-phenylethanol. Optimal conditions for immobilization have been investigated through a response surface methodology (RSM) and artificial neural network (ANN). The coefficient of determination (R(2)) and the root mean square error (RMSE) values between the calculated and estimated responses were respectively equal to 0.99 and 0.06 for the ANN training set, 0.97 and 0.2 for the ANN testing set, and 0.94 and 0.4 for the RSM training set. Both models provided good quality predictions, yet the ANN showed a clear superiority over RSM for both data fitting and estimation capabilities.

  8. Application of concrete surfaces as novel substrate for immobilization of TiO2 nano powder in photocatalytic treatment of phenolic water.

    Science.gov (United States)

    Delnavaz, Mohammad; Ayati, Bita; Ganjidoust, Hossein; Sanjabi, Sohrab

    2015-01-01

    In this study, concrete application as a substrate for TiO2 nano powder immobilization in heterogeneous photocatalytic process was evaluated. TiO2 immobilization on the pervious concrete surface was done by different procedures containing slurry method (SM), cement mixed method (CMM) and different concrete sealer formulations. Irradiation of TiO2 was prepared by UV-A and UV-C lamps. Phenolic wastewater was selected as a pollutant and efficiency of the process was determined in various operation conditions including influent phenol concentration, pH, TiO2 concentration, immobilization method and UV lamp intensity. The removal efficiency of photocatalytic process in 4 h irradiation time and phenol concentration ranges of 25-500 mg/L was more than 80 %. Intermediates were identified by GC/Mass and spectrophotometric analysis. According to the results, photocatalytic reactions followed the pseudo-first-order kinetics and can effectively treate phenol under optimal conditions.

  9. Spectroelectrochemical study of the [NiFe] hydrogenase from Desulfovibrio vulgaris Miyazaki F in solution and immobilized on biocompatible gold surfaces.

    Science.gov (United States)

    Millo, Diego; Pandelia, Maria-Eirini; Utesch, Tillmann; Wisitruangsakul, Nattawadee; Mroginski, Maria A; Lubitz, Wolfgang; Hildebrandt, Peter; Zebger, Ingo

    2009-11-19

    The catalytic cycle of the anaerobic [NiFe] hydrogenase from Desulfovibrio vulgaris Miyazaki F (DvMF) both in solution and immobilized on an Au electrode was studied by IR spectroscopic and electrochemical methods. IR spectroelectrochemistry in solution at different pH values allows the identification of the various redox-states of the active site and the determination of the midpoint potentials, as well as their acid-base equilibria. The spectroscopic characterization was based on the unique marker bands of the CN and CO stretching modes of the Ni-Fe center and served as reference for the surface-enhanced IR absorption (SEIRA) study of the immobilized enzyme. Using structural models of hydrogenases from DvMF and Desulfovibrio gigas , dipole moment calculations were carried out to guide the immobilization strategy. In view of the high dipole moment of about 1100 D pointing through the negatively charged area surrounding the distal [FeS] cluster, the Au electrode was coated by a self-assembled monolayer of amino-terminated mercaptanes which, due to the positively charged head groups, permit a durable electrostatic binding of the protein. SEIRA spectroscopy revealed a structurally and functionally intact active site as demonstrated by the reversible activation and inactivation under hydrogen and argon, respectively. Cyclic voltammetry on the immobilized enzyme demonstrate a reversible anaerobic inactivation upon changing the applied potential. The "switch" potential (E(switch)) associated with the reductive reactivation was determined to be -33 mV (vs normal hydrogen electrode). However, the catalytic current decreased on the time scale of hours during continuous cycling. SEIRA experiments demonstrate that the loss of catalytic activity is not due to protein desorption but is rather related to a slow degradation of the active site, possibly initiated by the attack of reactive species electrochemically generated from residual traces of oxygen in solution.

  10. Immobilization of Rhizobial Exopolysaccharides and Nod Factors Provides a Novel Platform for Interaction with Proteins

    DEFF Research Database (Denmark)

    Hjuler, Christian Toftegaard

    if LysM-RLKs from the model legumes L. japonicus and M. truncatula were able to bind Nod factors from rhizobia that they do not naturally form symbiosis with. These data could be useful in exploring the exact nature of the symbiosis. For this experiment proof of concept was obtained, but further work...

  11. Osteostatin improves the osteogenic activity of fibroblast growth factor-2 immobilized in Si-doped hydroxyapatite in osteoblastic cells.

    Science.gov (United States)

    Lozano, Daniel; Feito, María José; Portal-Núñez, Sergio; Lozano, Rosa María; Matesanz, María Concepción; Serrano, María Concepción; Vallet-Regí, María; Portolés, María Teresa; Esbrit, Pedro

    2012-07-01

    Si-doped hydroxyapatite (Si-HA) is a suitable ceramic for the controlled release of agents to improve bone repair. We recently showed that parathyroid hormone-related protein (PTHrP) (107-111) (osteostatin) has remarkable osteogenic features in various in vitro and in vivo systems. Fibroblast growth factor (FGF)-2 modulates osteoblastic function and induces angiogenesis, and can promote osteoblast adhesion and proliferation after immobilization on Si-HA. In the present study we examined whether osteostatin might improve the biological efficacy of FGF-2-coated Si-HA in osteoblastic MC3T3-E1 cells in vitro. We found that Si-HA/FGF-2 in the presence or absence of osteostatin (100 nM) similarly increased cell growth (by about 50%). However, addition of the latter peptide to Si-HA/FGF-2 significantly enhanced gene expression of Runx2, osteocalcin, vascular endothelial growth factor (VEGF) and the VEGF receptors 1 and 2, without significantly affecting that of FGF receptors in these cells. Moreover, secreted VEGF in the MC3T3-E1 cell conditioned medium, which induced the proliferation of pig endothelial-like cells, was also enhanced by these combined factors. The synergistic action of osteostatin and Si-HA/FGF-2 on the VEGF system was abrogated by a mitogen-activated protein kinase inhibitor (U0126) and by the calcium antagonist verapamil. This action was related to an enhancement of alkaline phosphatase activity and matrix mineralization in MC3T3-E1 cells, and also in primary human osteoblastic cells. These in vitro data show that osteostatin increases the osteogenic efficacy of a Si-HA/FGF-2 biomaterial by a mechanism involving mitogen-activated protein kinases and intracellular Ca(2+). These findings provide an attractive strategy for bone tissue engineering. Copyright © 2012 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

  12. Multiple epitope presentation and surface density control enabled by chemoselective immobilization lead to enhanced performance in IgE-binding fingerprinting on peptide microarrays.

    Science.gov (United States)

    Gori, Alessandro; Cretich, Marina; Vanna, Renzo; Sola, Laura; Gagni, Paola; Bruni, Giulia; Liprino, Marta; Gramatica, Furio; Burastero, Samuele; Chiari, Marcella

    2017-08-29

    Multiple ligand presentation is a powerful strategy to enhance the affinity of a probe for its corresponding target. A promising application of this concept lies in the analytical field, where surface immobilized probes interact with their corresponding targets in the context of complex biological samples. Here we investigate the effect of multiple epitope presentation (MEP) in the challenging context of IgE-detection in serum samples using peptide microarrays, and evaluate the influence of probes surface density on the assay results. Using the milk allergen alpha-lactalbumin as a model, we have synthesized three immunoreactive epitope sequences in a linear, branched and tandem form and exploited a chemoselective click strategy (CuAAC) for their immobilization on the surface of two biosensors, a microarray and an SPR chip both modified with the same clickable polymeric coating. We first demonstrated that a fine tuning of the surface peptide density plays a crucial role to fully exploit the potential of oriented and multiple peptide display. We then compared the three multiple epitope presentations in a microarray assay using sera samples from milk allergic patients, confirming that a multiple presentation, in particular that of the tandem construct, allows for a more efficient characterization of IgE-binding fingerprints at a statistically significant level. To gain insights on the binding parameters that characterize antibody/epitopes affinity, we selected the most reactive epitope of the series (LAC1) and performed a Surface Plasmon Resonance Imaging (SPRi) analysis comparing different epitope architectures (linear versus branched versus tandem). We demonstrated that the tandem peptide provides an approximately twofold increased binding capacity with respect to the linear and branched peptides, that could be attributed to a lower rate of dissociation (K d ). Copyright © 2017 Elsevier B.V. All rights reserved.

  13. Covalent co-immobilization of heparin/laminin complex that with different concentration ratio on titanium surface for selectively direction of platelets and vascular cells behavior

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Jian; Chen, Yuan; Liu, Tao; Wang, Xue; Liu, Yang; Wang, Yuan; Chen, Junying, E-mail: chenjy@263.net; Huang, Nan

    2014-10-30

    Highlights: • Extracellular matrix inspired surface modification with fibronectin, heparin and VEGF to construct a favorable microenvironment for selectively anticoagulant and promote endothelialization. • Take the advantage of specific intermolecular interaction, the bioactivity of above biomolecules was more efficiently maintained in compared with the common used covalent immobilization method. • Poly-l-lysine was used as a novel interlayer for surface amination, and in comparison, PLL coating was more feasible and the degradation product had no harm to human body. - Abstract: Surface biofunctional modification of coronary artery stent to improve the hemocompatibility and selectively accelerate endothelium regeneration but prevent restenosis have been become a new hotspot. For this, a novel method was developed in this work by co-immobilization of Ln and heparin complex on poly-L-lysine modified Ti surface. Take the advantage of the specific interaction between Ln and heparin, Ln and heparin complexes with different concentration ratios were set up for creating different exposure density of these two types of biomolecules. According to biocompatibility evaluation results, the Hep/Ln complexes modified surface displayed less platelet adhesion and activation. Especially, on L(150)H and L(200)H surface, the AT III binding quantity, APTT value and anti-coagulation property of modified surface were significantly promoted. Furthermore, the adherent density and proliferation activity of ECs and EPCs were positively correlated with Ln concentration. Notably, the proliferation of both ECs and EPCs on L(100)H, L(150)H and L(200)H surface were greatly promoted. Another hand, the proliferation activity of SMCs was significantly inhibited on Hep/Ln modified surfaces, which was considered mainly due to the inhibitory effect of heparin to SMCs. According to the existing results, this study demonstrated that in a certain range of heparin and laminin concentration ratio

  14. Covalent Immobilization of Enoxacin onto Titanium Implant Surfaces for Inhibiting Multiple Bacterial Species Infection and In Vivo Methicillin-Resistant Staphylococcus aureus Infection Prophylaxis.

    Science.gov (United States)

    Nie, Bin'en; Long, Teng; Ao, Haiyong; Zhou, Jianliang; Tang, Tingting; Yue, Bing

    2017-01-01

    Infection is one of the most important causes of titanium implant failure in vivo A developing prophylactic method involves the immobilization of antibiotics, especially vancomycin, onto the surface of the titanium implant. However, these methods have a limited effect in curbing multiple bacterial infections due to antibiotic specificity. In the current study, enoxacin was covalently bound to an amine-functionalized Ti surface by use of a polyethylene glycol (PEG) spacer, and the bactericidal effectiveness was investigated in vitro and in vivo The titanium surface was amine functionalized with 3-aminopropyltriethoxysilane (APTES), through which PEG spacer molecules were covalently immobilized onto the titanium, and then the enoxacin was covalently bound to the PEG, which was confirmed by X-ray photoelectron spectrometry (XPS). A spread plate assay, confocal laser scanning microscopy (CLSM), and scanning electron microscopy (SEM) were used to characterize the antimicrobial activity. For the in vivo study, Ti implants were inoculated with methicillin-resistant Staphylococcus aureus (MRSA) and implanted into the femoral medullary cavity of rats. The degree of infection was assessed by radiography, micro-computed tomography, and determination of the counts of adherent bacteria 3 weeks after surgery. Our data demonstrate that the enoxacin-modified PEGylated Ti surface effectively prevented bacterial colonization without compromising cell viability, adhesion, or proliferation in vitro Furthermore, it prevented MRSA infection of the Ti implants in vivo Taken together, our results demonstrate that the use of enoxacin-modified Ti is a potential approach to the alleviation of infections of Ti implants by multiple bacterial species. Copyright © 2016 American Society for Microbiology.

  15. Enhanced starch hydrolysis using α-amylase immobilized on cellulose ultrafiltration affinity membrane.

    Science.gov (United States)

    Konovalova, Viktoriia; Guzikevich, Kateryna; Burban, Anatoliy; Kujawski, Wojciech; Jarzynka, Karolina; Kujawa, Joanna

    2016-11-05

    In order to prepare ultrafiltration membranes possessing biocatalytic properties, α-amylase has been immobilized on cellulose membranes. Enzyme immobilization was based on a covalent bonding between chitosan and a surface of cellulose membrane, followed by an attachment of Cibacron Blue F3G-A dye as affinity ligand. Various factors affecting the immobilization process, such as enzyme concentration, pH of modifying solution, zeta-potential of membrane surface, and stability of immobilized enzyme were studied. The applicability of immobilized α-amylase has been investigated in ultrafiltration processes. The immobilization of α-amylase on membrane surface allows to increase the value of mass transfer coefficient and to decrease the concentration polarization effect during ultrafiltration of starch solutions. The enzyme layer on the membrane surface prevents a rapid increase of starch concentration due to the amylase hydrolysis of starch in the boundary layer. The presented affinity immobilization technique allows also for the regeneration of membranes from inactivated enzyme. Copyright © 2016 Elsevier Ltd. All rights reserved.

  16. Immobilization of Lipases on Alkyl Silane Modified Magnetic Nanoparticles: Effect of Alkyl Chain Length on Enzyme Activity

    Science.gov (United States)

    Wang, Jiqian; Meng, Gang; Tao, Kai; Feng, Min; Zhao, Xiubo; Li, Zhen; Xu, Hai; Xia, Daohong; Lu, Jian R.

    2012-01-01

    Background Biocatalytic processes often require a full recycling of biocatalysts to optimize economic benefits and minimize waste disposal. Immobilization of biocatalysts onto particulate carriers has been widely explored as an option to meet these requirements. However, surface properties often affect the amount of biocatalysts immobilized, their bioactivity and stability, hampering their wide applications. The aim of this work is to explore how immobilization of lipases onto magnetite nanoparticles affects their biocatalytic performance under carefully controlled surface modification. Methodology/Principal Findings Magnetite nanoparticles, prepared through a co-precipitation method, were coated with alkyl silanes of different alkyl chain lengths to modulate their surface hydrophobicity. Candida rugosa lipase was then directly immobilized onto the modified nanoparticles through hydrophobic interaction. Enzyme activity was assessed by catalytic hydrolysis of p-nitrophenyl acetate. The activity of immobilized lipases was found to increase with increasing chain length of the alkyl silane. Furthermore, the catalytic activities of lipases immobilized on trimethoxyl octadecyl silane (C18) modified Fe3O4 were a factor of 2 or more than the values reported from other surface immobilized systems. After 7 recycles, the activities of the lipases immobilized on C18 modified nanoparticles retained 65%, indicating significant enhancement of stability as well through hydrophobic interaction. Lipase immobilized magnetic nanoparticles facilitated easy separation and recycling with high activity retaining. Conclusions/Significance The activity of immobilized lipases increased with increasing alkyl chain length of the alkyl trimethoxy silanes used in the surface modification of magnetite nanoparticles. Lipase stability was also improved through hydrophobic interaction. Alkyl silane modified magnetite nanoparticles are thus highly attractive carriers for enzyme immobilization

  17. Immobilization of lipases on alkyl silane modified magnetic nanoparticles: effect of alkyl chain length on enzyme activity.

    Directory of Open Access Journals (Sweden)

    Jiqian Wang

    Full Text Available BACKGROUND: Biocatalytic processes often require a full recycling of biocatalysts to optimize economic benefits and minimize waste disposal. Immobilization of biocatalysts onto particulate carriers has been widely explored as an option to meet these requirements. However, surface properties often affect the amount of biocatalysts immobilized, their bioactivity and stability, hampering their wide applications. The aim of this work is to explore how immobilization of lipases onto magnetite nanoparticles affects their biocatalytic performance under carefully controlled surface modification. METHODOLOGY/PRINCIPAL FINDINGS: Magnetite nanoparticles, prepared through a co-precipitation method, were coated with alkyl silanes of different alkyl chain lengths to modulate their surface hydrophobicity. Candida rugosa lipase was then directly immobilized onto the modified nanoparticles through hydrophobic interaction. Enzyme activity was assessed by catalytic hydrolysis of p-nitrophenyl acetate. The activity of immobilized lipases was found to increase with increasing chain length of the alkyl silane. Furthermore, the catalytic activities of lipases immobilized on trimethoxyl octadecyl silane (C18 modified Fe(3O(4 were a factor of 2 or more than the values reported from other surface immobilized systems. After 7 recycles, the activities of the lipases immobilized on C18 modified nanoparticles retained 65%, indicating significant enhancement of stability as well through hydrophobic interaction. Lipase immobilized magnetic nanoparticles facilitated easy separation and recycling with high activity retaining. CONCLUSIONS/SIGNIFICANCE: The activity of immobilized lipases increased with increasing alkyl chain length of the alkyl trimethoxy silanes used in the surface modification of magnetite nanoparticles. Lipase stability was also improved through hydrophobic interaction. Alkyl silane modified magnetite nanoparticles are thus highly attractive carriers for

  18. Immobilization of lipases on alkyl silane modified magnetic nanoparticles: effect of alkyl chain length on enzyme activity.

    Science.gov (United States)

    Wang, Jiqian; Meng, Gang; Tao, Kai; Feng, Min; Zhao, Xiubo; Li, Zhen; Xu, Hai; Xia, Daohong; Lu, Jian R

    2012-01-01

    Biocatalytic processes often require a full recycling of biocatalysts to optimize economic benefits and minimize waste disposal. Immobilization of biocatalysts onto particulate carriers has been widely explored as an option to meet these requirements. However, surface properties often affect the amount of biocatalysts immobilized, their bioactivity and stability, hampering their wide applications. The aim of this work is to explore how immobilization of lipases onto magnetite nanoparticles affects their biocatalytic performance under carefully controlled surface modification. Magnetite nanoparticles, prepared through a co-precipitation method, were coated with alkyl silanes of different alkyl chain lengths to modulate their surface hydrophobicity. Candida rugosa lipase was then directly immobilized onto the modified nanoparticles through hydrophobic interaction. Enzyme activity was assessed by catalytic hydrolysis of p-nitrophenyl acetate. The activity of immobilized lipases was found to increase with increasing chain length of the alkyl silane. Furthermore, the catalytic activities of lipases immobilized on trimethoxyl octadecyl silane (C18) modified Fe(3)O(4) were a factor of 2 or more than the values reported from other surface immobilized systems. After 7 recycles, the activities of the lipases immobilized on C18 modified nanoparticles retained 65%, indicating significant enhancement of stability as well through hydrophobic interaction. Lipase immobilized magnetic nanoparticles facilitated easy separation and recycling with high activity retaining. The activity of immobilized lipases increased with increasing alkyl chain length of the alkyl trimethoxy silanes used in the surface modification of magnetite nanoparticles. Lipase stability was also improved through hydrophobic interaction. Alkyl silane modified magnetite nanoparticles are thus highly attractive carriers for enzyme immobilization enabling efficient enzyme recovery and recycling.

  19. Development and Validation of an On-Line Water Toxicity Sensor with Immobilized Luminescent Bacteria for On-Line Surface Water Monitoring

    Directory of Open Access Journals (Sweden)

    Marjolijn Woutersen

    2017-11-01

    Full Text Available Surface water used for drinking water production is frequently monitored in The Netherlands using whole organism biomonitors, with for example Daphnia magna or Dreissena mussels, which respond to changes in the water quality. However, not all human-relevant toxic compounds can be detected by these biomonitors. Therefore, a new on-line biosensor has been developed, containing immobilized genetically modified bacteria, which respond to genotoxicity in the water by emitting luminescence. The performance of this sensor was tested under laboratory conditions, as well as under field conditions at a monitoring station along the river Meuse in The Netherlands. The sensor was robust and easy to clean, with inert materials, temperature control and nutrient feed for the reporter organisms. The bacteria were immobilized in sol-gel on either an optical fiber or a glass slide and then continuously exposed to water. Since the glass slide was more sensitive and robust, only this setup was used in the field. The sensor responded to spikes of genotoxic compounds in the water with a minimal detectable concentration of 0.01 mg/L mitomycin C in the laboratory and 0.1 mg/L mitomycin C in the field. With further optimization, which should include a reduction in daily maintenance, the sensor has the potential to become a useful addition to the currently available biomonitors.

  20. Response surface optimization for the transesterification of karanja oil using immobilized whole cells of Rhizopus oryzae in n-hexane system

    Energy Technology Data Exchange (ETDEWEB)

    Ganesan, Devanesan; Rajendran, Aravindan; Thangavelu, Viruthagiri [Annamalai University, Department of Chemical Engineering, Faculty of Engineering and Technology, Biochemical Engineering Laboratory, Annamalai Nagar, Tamil Nadu (India)

    2012-03-15

    Non-edible oils represent one of the most viable alternative feed stocks for the production of large volumes of biodiesel at cheaper cost in tropical countries. The objective of the present study is to investigate the ability of the immobilized whole cells of Rhizopus oryzae MTCC 262 to catalyze the biodiesel production from karanja oil in n-hexane system. Response surface methodology was employed to evaluate the effects of synthesis parameters, such as molar ratio of oil to alcohol, reaction temperature and reaction time on percentage biodiesel (methyl esters) yield. Transesterification was performed in shake flasks containing immobilized cells in the reaction mixture with 10% oil weight of n-hexane. The quadratic effects of molar ratio of oil to alcohol and reaction time proved to be the significant at 1% and 5% levels, respectively. The optimum synthesis conditions were found to be: molar ratio of oil to alcohol 1:2.73, reaction temperature 41.39 C and reaction time 73.97 h. Biodiesel yield (methyl ester) was 75.98 (wt.%) under the optimal conditions and the subsequent verification experiments with biodiesel yield of 78.0 (wt.%) confirmed the validity of the proposed model. (orig.)

  1. Linker-free covalent immobilization of heparin, SDF-1α, and CD47 on PTFE surface for antithrombogenicity, endothelialization and anti-inflammation.

    Science.gov (United States)

    Gao, Ang; Hang, Ruiqiang; Li, Wan; Zhang, Wei; Li, Penghui; Wang, Guomin; Bai, Long; Yu, Xue-Feng; Wang, Huaiyu; Tong, Liping; Chu, Paul K

    2017-09-01

    Small-diameter vascular grafts made of biomedical polytetrafluoroethylene (PTFE) suffer from the poor long-term patency rate originating from thrombosis and intimal hyperplasia, which can be ascribed to the insufficient endothelialization and chronic inflammation of the materials. Hence, bio-functionalization of PTFE grafts is highly desirable to circumvent these disadvantages. In this study, a versatile "implantation-incubation" approach in which the biomedical PTFE is initially modified by plasma immersion ion implantation (PIII) is described. After the N 2 PIII treatment, the surface of biomedical PTFE is roughened with nanostructures and more importantly, the abundant free radicals generated underneath the surface continuously migrate to the surface and react with environmental molecules. Taking advantage of this mechanism, various biomolecules with different functions can be steadily immobilized on the surface of PTFE by simple solution immersion. As examples, three typical biomolecules, heparin, SDF-1α, and CD47, are covalently grafted onto the PTFE. In addition to retaining the bioactivity, the surface-functionalized PTFE exhibits reduced thrombogenicity, facilitates the recruitment of endothelial progenitor cells, and even alleviates the inflammatory immune responses of monocytes-macrophages and is thus promising to the development of small-diameter prosthetic vascular grafts with good long-term patency. Copyright © 2017 Elsevier Ltd. All rights reserved.

  2. Self-Immobilization of Car9 Fusion Proteins within High Surface Area Silica Sol-Gels and Dynamic Control of Protein Release.

    Science.gov (United States)

    Yang, Wenlan; Hellner, Brittney; Baneyx, François

    2016-10-19

    Protein entrapment within silica matrices during sol-gel formation is an effective way of producing biocatalysts with high load, activity retention, and minimal leaching. On the other hand, mesoporous silica materials have been favored for diffusional control of protein delivery because of their regular pore size and morphology and in spite of the drawback of requiring post-synthesis loading with cargo proteins. Here, we describe a hybrid technology in which fusion of the silica-binding Car9 dodecapeptide to model fluorescent proteins allows for their simultaneous entrapment and surface immobilization within sol-gel monoliths that can be fabricated in air and oil phases. Spherical particles produced by injecting a mixture of silicic acid and Car9-tagged proteins in silicone oil exhibit high surface area (>400 m 2 /g), 15-nm-diameter mean pore size and homogeneous protein loading. Incubation in arginine-containing buffer disrupts the interaction between Car9 extensions and silica surfaces and triggers the continuous or discontinuous (on/off) release of cargo proteins with pH-tunable kinetics. This simple approach for producing hybrid silica materials that stably encapsulate and release one or more Car9-tagged proteins in a single step may prove useful for applications requiring dynamic control of protein concentration.

  3. Covalent immobilization of metal–organic frameworks onto the surface of nylon—a new approach to the functionalization and coloration of textiles

    Science.gov (United States)

    Yu, Ming; Li, Wanxin; Wang, Ziqiang; Zhang, Bowu; Ma, Hongjuan; Li, Linfan; Li, Jingye

    2016-01-01

    The prevention of refractory organic pollution caused by conventional dyeing and the development of new fabrics with various functions are two issues to be solved urgently in the field of textile fabrication. Here, we report a new environmentally friendly route for the simultaneous coloration and functionalization of textiles by the covalent immobilization of a metal–organic framework, Cr-based MIL-101(Cr), onto the surfaces of nylon fabrics by co-graft polymerization with 2-hydroxyethyl acrylate initiated by γ-ray irradiation. The Cr(III) clusters color the nylon fabric, and the color intensity varies with the MIL-101 content, providing a “green” textile coloration method that is different from conventional dyeing processes. An X-ray diffraction (XRD) analysis shows that the nanoporous structure of the original MIL-101 particles is retained during radiation-induced graft polymerization. Numerous nanopores are introduced onto the surface of the nylon fabric, which demonstrated better sustained-release-of-aroma performance versus pristine nylon fabric in tests. The modified fabrics exhibit laundering durability, with MIL-101 nanoparticles intact on the nylon surface after 30 h of dry cleaning. PMID:26948405

  4. Hybrid response surface methodology-genetic algorithm optimization of ultrasound-assisted transesterification of waste oil catalysed by immobilized lipase on mesoporous silica/iron oxide magnetic core-shell nanoparticles.

    Science.gov (United States)

    Karimi, Mahmoud; Keyhani, Alireza; Akram, Asadolah; Rahman, Masoud; Jenkins, Bryan; Stroeve, Pieter

    2013-01-01

    The production ofbiodiesel by transesterification of waste cooking oil (WCO) to partially substitute petroleum diesel is one of the measures for solving the twin problems of environment pollution and energy demand. An environmentally benign process for the enzymatic transesterification using immobilized lipase has attracted considerable attention for biodiesel production. Here, a superparamagnetic, high surface area substrate for lipase immobilization is evaluated. These immobilization substrates are composed of mesoporous silica/superparamagnetic iron oxide core-shell nanoparticles. The effects of methanol ratio to WCO, lipase concentration, water content and reaction time on the synthesis of biodiesel were analysed by utilizing the response surface methodology (RSM). A quadratic response surface equation for calculating fatty acid methyl ester (FAME) content as the objective function was established based on experimental data obtained in accordance with the central composite design. The RSM-based model was then used as the fitness function for genetic algorithm (GA) to optimize its input space. Hybrid RSM-GA predicted the maximum FAME content (91%) at the optimum level of medium variables: methanol ratio to WCO, 4.34; lipase content, 43.6%; water content, 10.22%; and reaction time, 6h. Moreover, the immobilized lipase could be used for four times without considerable loss of the activity.

  5. Anhydride-functional silane immobilized onto titanium surfaces induces osteoblast cell differentiation and reduces bacterial adhesion and biofilm formation

    Energy Technology Data Exchange (ETDEWEB)

    Godoy-Gallardo, Maria, E-mail: maria.godoy.gallardo@upc.edu [Biomaterials, Biomechanics and Tissue Engineering Group, Department of Materials Science and Metallurgy, Technical University of Catalonia (UPC), ETSEIB, Av. Diagonal 647, 08028 Barcelona (Spain); Centre for Research in NanoEngineering (CRNE) — UPC, C/ Pascual i Vila 15, 08028 Barcelona (Spain); Guillem-Marti, Jordi, E-mail: jordi.guillem.marti@upc.edu [Biomaterials, Biomechanics and Tissue Engineering Group, Department of Materials Science and Metallurgy, Technical University of Catalonia (UPC), ETSEIB, Av. Diagonal 647, 08028 Barcelona (Spain); Centre for Research in NanoEngineering (CRNE) — UPC, C/ Pascual i Vila 15, 08028 Barcelona (Spain); Sevilla, Pablo, E-mail: psevilla@euss.es [Department of Mechanics, Escola Universitària Salesiana de Sarrià (EUSS), C/ Passeig de Sant Bosco, 42, 08017 Barcelona (Spain); Manero, José M., E-mail: jose.maria.manero@upc.edu [Biomaterials, Biomechanics and Tissue Engineering Group, Department of Materials Science and Metallurgy, Technical University of Catalonia (UPC), ETSEIB, Av. Diagonal 647, 08028 Barcelona (Spain); Centre for Research in NanoEngineering (CRNE) — UPC, C/ Pascual i Vila 15, 08028 Barcelona (Spain); Gil, Francisco J., E-mail: francesc.xavier.gil@upc.edu [Biomaterials, Biomechanics and Tissue Engineering Group, Department of Materials Science and Metallurgy, Technical University of Catalonia (UPC), ETSEIB, Av. Diagonal 647, 08028 Barcelona (Spain); Centre for Research in NanoEngineering (CRNE) — UPC, C/ Pascual i Vila 15, 08028 Barcelona (Spain); and others

    2016-02-01

    Bacterial infection in dental implants along with osseointegration failure usually leads to loss of the device. Bioactive molecules with antibacterial properties can be attached to titanium surfaces with anchoring molecules such as silanes, preventing biofilm formation and improving osseointegration. Properties of silanes as molecular binders have been thoroughly studied, but research on the biological effects of these coatings is scarce. The aim of the present study was to determine the in vitro cell response and antibacterial effects of triethoxysilypropyl succinic anhydride (TESPSA) silane anchored on titanium surfaces. X-ray photoelectron spectroscopy confirmed a successful silanization. The silanized surfaces showed no cytotoxic effects. Gene expression analyses of Sarcoma Osteogenic (SaOS-2) osteoblast-like cells cultured on TESPSA silanized surfaces reported a remarkable increase of biochemical markers related to induction of osteoblastic cell differentiation. A manifest decrease of bacterial adhesion and biofilm formation at early stages was observed on treated substrates, while favoring cell adhesion and spreading in bacteria–cell co-cultures. Surfaces treated with TESPSA could enhance a biological sealing on implant surfaces against bacteria colonization of underlying tissues. Furthermore, it can be an effective anchoring platform of biomolecules on titanium surfaces with improved osteoblastic differentiation and antibacterial properties. - Highlights: • TESPSA silane induces osteoblast differentiation. • TESPSA reduces bacterial adhesion and biofilm formation. • TESPSA is a promising anchoring platform of biomolecules onto titanium.

  6. Anhydride-functional silane immobilized onto titanium surfaces induces osteoblast cell differentiation and reduces bacterial adhesion and biofilm formation

    International Nuclear Information System (INIS)

    Godoy-Gallardo, Maria; Guillem-Marti, Jordi; Sevilla, Pablo; Manero, José M.; Gil, Francisco J.

    2016-01-01

    Bacterial infection in dental implants along with osseointegration failure usually leads to loss of the device. Bioactive molecules with antibacterial properties can be attached to titanium surfaces with anchoring molecules such as silanes, preventing biofilm formation and improving osseointegration. Properties of silanes as molecular binders have been thoroughly studied, but research on the biological effects of these coatings is scarce. The aim of the present study was to determine the in vitro cell response and antibacterial effects of triethoxysilypropyl succinic anhydride (TESPSA) silane anchored on titanium surfaces. X-ray photoelectron spectroscopy confirmed a successful silanization. The silanized surfaces showed no cytotoxic effects. Gene expression analyses of Sarcoma Osteogenic (SaOS-2) osteoblast-like cells cultured on TESPSA silanized surfaces reported a remarkable increase of biochemical markers related to induction of osteoblastic cell differentiation. A manifest decrease of bacterial adhesion and biofilm formation at early stages was observed on treated substrates, while favoring cell adhesion and spreading in bacteria–cell co-cultures. Surfaces treated with TESPSA could enhance a biological sealing on implant surfaces against bacteria colonization of underlying tissues. Furthermore, it can be an effective anchoring platform of biomolecules on titanium surfaces with improved osteoblastic differentiation and antibacterial properties. - Highlights: • TESPSA silane induces osteoblast differentiation. • TESPSA reduces bacterial adhesion and biofilm formation. • TESPSA is a promising anchoring platform of biomolecules onto titanium.

  7. Surface water quality assessment using factor analysis

    African Journals Online (AJOL)

    2006-01-16

    Jan 16, 2006 ... factors extracted by the Centroid method, rotated by Varimax rotation (Ahmed et al., 2005). Calculated eigenvalues, per cent. Figure 1. Sketch map of water quality monitoring stations in Buyuk. Menderes Basin in Turkey. TABLE 1. Descriptive statistics of water quality data under low-flow conditions. Variable.

  8. Strong and Reversible Monovalent Supramolecular Protein Immobilization

    NARCIS (Netherlands)

    Young, Jacqui F.; Nguyen, Hoang D.; Yang, Lanti; Huskens, Jurriaan; Jonkheijm, Pascal; Brunsveld, Luc

    2010-01-01

    Proteins with an iron clasp: Site-selective incorporation of a ferrocene molecule into a protein allows for easy, strong, and reversible supramolecular protein immobilization through a selective monovalent interaction of the ferrocene with a cucurbit[7]uril immobilized on a gold surface. The

  9. Factors influencing graphene growth on metal surfaces

    International Nuclear Information System (INIS)

    Loginova, E; Bartelt, N C; McCarty, K F; Feibelman, P J

    2009-01-01

    Graphene forms from a relatively dense, tightly bound C-adatom gas when elemental C is deposited on or segregates to the Ru(0001) surface. Nonlinearity of the graphene growth rate with C-adatom density suggests that growth proceeds by addition of C atom clusters to the graphene edge. The generality of this picture has now been studied by use of low-energy electron microscopy (LEEM) to observe graphene formation when Ru(0001) and Ir(111) surfaces are exposed to ethylene. The finding that graphene growth velocities and nucleation rates on Ru have precisely the same dependence on adatom concentration as for elemental C deposition implies that hydrocarbon decomposition only affects graphene growth through the rate of adatom formation. For ethylene, that rate decreases with increasing adatom concentration and graphene coverage. Initially, graphene growth on Ir(111) is like that on Ru: the growth velocity is the same nonlinear function of adatom concentration (albeit with much smaller equilibrium adatom concentrations, as we explain with DFT calculations of adatom formation energies). In the later stages of growth, graphene crystals that are rotated relative to the initial nuclei nucleate and grow. The rotated nuclei grow much faster. This difference suggests firstly, that the edge-orientation of the graphene sheets relative to the substrate plays an important role in the growth mechanism, and secondly, that attachment of the clusters to the graphene is the slowest step in cluster addition, rather than formation of clusters on the terraces.

  10. Chondrogenesis and hypertrophy in response to aggregate behaviors of human mesenchymal stem cells on a dendrimer-immobilized surface.

    Science.gov (United States)

    Wongin, Sopita; Ogawa, Yuuki; Kim, Mee-Hae; Viravaidya-Pasuwat, Kwanchanok; Kino-Oka, Masahiro

    2017-08-01

    To investigate the behaviors of aggregates of human mesenchymal stem cells (hMSCs) on chondrogenesis and chondrocyte hypertrophy using spatiotemporal expression patterns of chondrogenic (type II collagen) and hypertrophic (type X collagen) markers during chondrogenesis. hMSCs were cultured on either a polystyrene surface or polyamidoamine dendrimer surface with a fifth generation (G5) dendron structure in chondrogenic medium and growth medium. At day 7, cell aggregates without stress fibers formed on the G5 surface and triggered differentiation of hMSCs toward the chondrogenic fate, as indicated by type II collagen being observed while type X collagen was undetectable. In contrast, immunostaining of hMSCs cultured on polystyrene, which exhibited abundant stress fibers and did not form aggregates, revealed no evidence of either type II and or type X collagen. At day 21, the morphological changes of the cell aggregates formed on the G5 surface were suppressed as a result of stress fiber formation. Type II collagen was observed throughout the aggregates whereas type X collagen was detected only at the basal side of the aggregates. Change of cell aggregate behaviors derived from G5 surface alone regulated chondrogenesis and hypotrophy, and this was enhanced by chondrogenic medium. Incubation of hMSCs affects the expression of type II and X collagens via effects on cell aggregate behavior and stress fiber formation.

  11. Osseous response on linear and cyclic RGD-peptides immobilized on titanium surfaces in vitro and in vivo.

    Science.gov (United States)

    Heller, M; Kumar, V V; Pabst, A; Brieger, J; Al-Nawas, B; Kämmerer, P W

    2018-02-01

    Biomimetic surface modifications of titanium (Ti) implants using the Arg-Gly-Asp-sequence (RGD) are promising to accelerate bone healing in cases of medical implants. Therefore, we compared the impact of linear and cyclic RGD (l- and c-RGD) covalently coupled onto Ti surfaces on the osseous response in vitro and in vivo. In vitro, osteoblasts' behavior on different surfaces (unmodified, amino-silanized [APTES], l- and c-RGD) was analysed regarding adhesion (fluorescence microscopy), proliferation (resazurin stain) and differentiation (reverse transcription polymerase chain reaction on alkaline phosphatase and osteocalcin). In vivo, osteosynthesis screws (unmodified n = 8, l-RGD n = 8, c-RGD n = 8) were inserted into the proximal tibiae of 12 rabbits and evaluated for bone growth parameters (bone implant contact [%] and vertical bone apposition [VBA;%]) at 3 and 6 weeks. In vitro, c- as well as l-RGD surfaces stimulated osteoblasts' adherence, proliferation and differentiation in a similar manner, with only subtle evidence of superiority of the c-RGD modifications. In vivo, c-RGD-modifications led to a significantly increased VBA after 3 and 6 weeks. Thus, coating with c-RGD appears to play an important role influencing osteoblasts' behaviour in vitro but especially in vivo. These findings can be applied prospectively to implantable biomaterials with hypothetically improved survival and success rates. © 2017 Wiley Periodicals Inc. J Biomed Mater Res Part A: 106A: 419-427, 2018. © 2017 Wiley Periodicals, Inc.

  12. Kinetic analysis of inhibition of glucoamylase and active site mutants via chemoselective oxime immobilization of acarbose on SPR chip surfaces

    DEFF Research Database (Denmark)

    Sauer, Jørgen; Abou Hachem, Maher; Svensson, Birte

    2013-01-01

    We here report a quantitative study on the binding kinetics of inhibition of the enzyme glucoamylase and how individual active site amino acid mutations influence kinetics. To address this challenge, we have developed a fast and efficient method for anchoring native acarbose to gold chip surfaces...

  13. Functional immobilization of recombinant alkaline phosphatases bearing a glutamyl donor substrate peptide of microbial transglutaminase.

    Science.gov (United States)

    Kamiya, Noriho; Doi, Satoshi; Tanaka, Yusuke; Ichinose, Hirofumi; Goto, Masahiro

    2007-09-01

    Covalent and site-specific protein immobilization catalyzed by microbial transglutaminase (MTG) was investigated using recombinant Escherichia coli alkaline phosphatase (AP) tagged with a glutamyl donor substrate peptide (MLAQGS) of MTG. A polystyrene surface physically coated with beta-casein or bovine serum albumin (BSA) was employed as an MTG-specific surface displaying reactive lysine residues. MTG-mediated protein immobilization through catalytic epsilon-(gamma-glutamyl)lysine bond formation between the peptide tag of recombinant APs and beta-casein- or BSA-coated surface was verified by the detection of AP activity on the surface. It was found that the length and the insertion position of the peptide tag did not significantly affect the efficacy of enzymatic immobilization of the recombinant APs. On the other hand, pH and ionic strength in the reaction media had crucial effects on the immobilization yields. Interestingly, the optimum pH range of MTG-mediated protein immobilization differed markedly from that for an MTG-catalyzed reaction in aqueous solution. The results suggest that the concentration of reactive species due to electrostatic interaction between the enzyme-substrate intermediate and the protein-adsorbed surface is a key factor governing MTG catalysis at a solid surface.

  14. Preparation and Electrocatalytic Activity of Gold Nanoparticles Immobilized on the Surface of 4-Mercaptobenzoyl-Functionalized Multiwalled Carbon Nanotubes

    Science.gov (United States)

    2010-12-20

    the surface of CNTs are most probably the sites that deactivate transition metal catalysts.13 We have developed an efficient method to introduce...like precipitate was collected using suction filtration and washed with distilled water. The product was Soxhlet -extracted with water overnight to...anchoring sites for the uniform dispersion and stabilization of GNPs. TEM element mapping was used as an additional method to characterize the uniformity

  15. Voltammetric Detection of S100B Protein Using His-Tagged Receptor Domains for Advanced Glycation End Products (RAGE Immobilized onto a Gold Electrode Surface

    Directory of Open Access Journals (Sweden)

    Edyta Mikuła

    2014-06-01

    Full Text Available In this work we report on an electrochemical biosensor for the determination of the S100B protein. The His-tagged VC1 domains of Receptors for Advanced Glycation End (RAGE products used as analytically active molecules were covalently immobilized on a monolayer of a thiol derivative of pentetic acid (DPTA complex with Cu(II deposited on a gold electrode surface. The recognition processes between the RAGE VC1 domain and the S100B protein results in changes in the redox activity of the DPTA-Cu(II centres which were measured by Osteryoung square-wave voltammetry (OSWV. In order to verify whether the observed analytical signal originates from the recognition process between the His6–RAGE VC1 domains and the S100B protein, the electrode modified with the His6–RAGE C2 and His6–RAGE VC1 deleted domains which have no ability to bind S100B peptides were applied. The proposed biosensor was quite sensitive, with a detection limit of 0.52 pM recorded in the buffer solution. The presence of diluted human plasma and 10 nM Aβ1-40 have no influence on the biosensor performance.

  16. A simple fabrication of plasmonic surface-enhanced Raman scattering (SERS) substrate for pesticide analysis via the immobilization of gold nanoparticles on UF membrane

    Science.gov (United States)

    Hong, Jangho; Kawashima, Ayato; Hamada, Noriaki

    2017-06-01

    In this study, we developed a facile fabrication method to access a highly reproducible plasmonic surface enhanced Raman scattering substrate via the immobilization of gold nanoparticles on an Ultrafiltration (UF) membrane using a suction technique. This was combined with a simple and rapid analyte concentration and detection method utilizing portable Raman spectroscopy. The minimum detectable concentrations for aqueous thiabendazole standard solution and thiabendazole in orange extract are 0.01 μg/mL and 0.125 μg/g, respectively. The partial least squares (PLS) regression plot shows a good linear relationship between 0.001 and 100 μg/mL of analyte, with a root mean square error of prediction (RMSEP) of 0.294 and a correlation coefficient (R2) of 0.976 for the thiabendazole standard solution. Meanwhile, the PLS plot also shows a good linear relationship between 0.0 and 2.5 μg/g of analyte, with an RMSEP value of 0.298 and an R2 value of 0.993 for the orange peel extract. In addition to the detection of other types of pesticides in agricultural products, this highly uniform plasmonic substrate has great potential for application in various environmentally-related areas.

  17. Titanium surfaces immobilized with the major antimicrobial fragment FK-16 of human cathelicidin LL-37 are potent against multiple antibiotic-resistant bacteria.

    Science.gov (United States)

    Mishra, Biswajit; Wang, Guangshun

    2017-08-01

    Infections on implanted medical devices are a challenging problem, especially when bacteria form difficult-to-treat biofilms. Antimicrobial peptides are considered to be a solution due to their potency against antibiotic-resistant superbugs. Previously, the authors' laboratory demonstrated the prevention of staphylococcal biofilm formation in an animal catheter model by injecting merecidin (formerly known as 17BIPHE2), a peptide engineered based on the only human cathelicidin. This study documents an alternative solution via covalent immobilization of FK-16, amino acid sequence FKRIVQRIKDFLRNLV-amide, which corresponds to the major antimicrobial region (residues 17-32) of LL-37. FK-16 is superior to the longer peptide LL-37 in terms of synthesis cost and the shorter peptide KR-12 in terms of activity spectrum. Indeed, the FK16-coated titanium surface showed a broad-spectrum activity against the ESKAPE pathogens, including Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter species. It also demonstrated anti-adhesion and biofilm inhibition capabilities against both S. aureus and E. coli.

  18. Immobilized molybdenum–thiosemicarbazide Schiff base complex on the surface of magnetite nanoparticles as a new nanocatalyst for the epoxidation of olefins

    Energy Technology Data Exchange (ETDEWEB)

    Mohammadikish, M., E-mail: mohammadikish@yahoo.com [Department of Chemistry, Faculty of Sciences, Kermanshah Branch, Islamic Azad University, Kermanshah (Iran, Islamic Republic of); Masteri-Farahani, M.; Mahdavi, S. [Faculty of Chemistry, Kharazmi University, Tehran (Iran, Islamic Republic of)

    2014-03-15

    In this work, a new magnetically recoverable nanocatalyst was developed by immobilization of thiosemicarbazide ligand on the surface of silica coated magnetite nanoparticles (SCMNPs) through Schiff base condensation and followed complexation with MoO{sub 2}(acac){sub 2}. Characterization of the prepared nanocatalyst was performed with different physicochemical methods such as Fourier transform infrared (FT-IR) and atomic absorption spectroscopies, X-ray diffraction (XRD), vibrating sample magnetometry (VSM), thermogravimetric analysis (TGA), field-emission scanning electron microscopy (FE-SEM), and transmission electron microscopy (TEM). The prepared catalyst catalyzed the epoxidation of olefins and allyl alcohols with tert-butyl hydroperoxide (TBHP) and cumene hydroperoxide (CHP) quantitatively with excellent selectivity toward the corresponding epoxides under mild reaction conditions. - Highlights: • Silica coated magnetite nanoparticles were modified with a thiosemicarbazide-Schiff base ligand. • Complexation of the supported ligand with molybdenum resulted in preparation of a new hybrid nanomaterial. • The prepared hybrid nanomaterial acted as an efficient and reusable catalyst in the epoxidation of olefins.

  19. Effective immobilization of Ru(bpy)32+ by functional composite phosphomolybdic acid anion on an electrode surface for solid-state electrochemiluminescene to sensitive determination of NADH

    International Nuclear Information System (INIS)

    Li Yali; Yang Xiurong; Yang Fan; Wang Yingping; Zheng Peihua; Liu Xiaoxu

    2012-01-01

    Phosphomolybdic acid anion ([PMo 12 O 40 ] 3− ) was used for the immobilization of ruthenium(II) tris(bipyridine) (Ru(bpy) 3 2+ ) on an electrode surface to yield a sensitive solid-state electrogenerated chemiluminescence (ECL) sensor. [PMo 12 O 40 ] 3− anion in the prepared sensor had catalytic ability to the NADH oxidation. The ECL signal of the Ru(bpy) 3 2+ /[PMo 12 O 40 ] 3− film was about 3-fold enhancement than that for the Ru(bpy) 3 2+ /Nafion film to NADH determination. The resulting ECL sensor exhibited a wide linear range from 2.5 × 10 −7 to 5.0 × 10 −3 M (R = 0.99) with the detection limit of 1.67 × 10 −8 M (S/N = 3). In addition, it had good reproducibility and excellent long-term stability, and the relative average deviation was 0.77% of ECL intensity–time curve under continuous potential scanning for 21 cycles; after being used in two weeks, the sensor was able to keep over 90% activity toward 25 μM NADH. Fabrication of the ECL sensor by this method is simple and easy. Such superior properties will promote the application of polyoxometalates in fabricating sensors for using in electroanalytical and biochemical analysis.

  20. Functionalizable surface platform with reduced nonspecific protein adsorption from bull blood plasma - Material selection and protein immobilization optimization

    Czech Academy of Sciences Publication Activity Database

    Vaisocherová, Hana; Zhang, Z.; Yang, W.; Cao, Z.; Cheng, G.; Taylor, A. D.; Piliarik, Marek; Homola, Jiří; Jiang, S.

    2009-01-01

    Roč. 24, č. 7 (2009), s. 1924-1930 ISSN 0956-5663 R&D Projects: GA AV ČR KAN200670701 Institutional research plan: CEZ:AV0Z20670512 Keywords : Zwitterionic poly(carboxybetaine) * Functionalization * Ultra-low fouling Subject RIV: CD - Macromolecular Chemistry Impact factor: 5.429, year: 2009

  1. Functionalized Graphene Sheets As Immobilization Matrix for Fenugreek β-Amylase: Enzyme Kinetics and Stability Studies

    Science.gov (United States)

    Srivastava, Garima; Singh, Kritika; Talat, Mahe; Srivastava, Onkar Nath; Kayastha, Arvind M.

    2014-01-01

    β-Amylase finds application in food and pharmaceutical industries. Functionalized graphene sheets were customised as a matrix for covalent immobilization of Fenugreek β-amylase using glutaraldehyde as a cross-linker. The factors affecting the process were optimized using Response Surface Methodology based Box-Behnken design of experiment which resulted in 84% immobilization efficiency. Scanning and Transmission Electron Microscopy (SEM, TEM) and Fourier Tansform Infrared (FTIR) spectroscopy were employed for the purpose of characterization of attachment of enzyme on the graphene. The enzyme kinetic studies were carried out for obtaining best catalytic performance and enhanced reusability. Optimum temperature remained unchanged, whereas optimum pH showed shift towards acidic range for immobilized enzyme. Increase in thermal stability of immobilized enzyme and non-toxic nature of functionalized graphene can be exploited for production of maltose in food and pharmaceutical industries. PMID:25412079

  2. Functionalized graphene sheets as immobilization matrix for Fenugreek β-amylase: enzyme kinetics and stability studies.

    Directory of Open Access Journals (Sweden)

    Garima Srivastava

    Full Text Available β-Amylase finds application in food and pharmaceutical industries. Functionalized graphene sheets were customised as a matrix for covalent immobilization of Fenugreek β-amylase using glutaraldehyde as a cross-linker. The factors affecting the process were optimized using Response Surface Methodology based Box-Behnken design of experiment which resulted in 84% immobilization efficiency. Scanning and Transmission Electron Microscopy (SEM, TEM and Fourier Tansform Infrared (FTIR spectroscopy were employed for the purpose of characterization of attachment of enzyme on the graphene. The enzyme kinetic studies were carried out for obtaining best catalytic performance and enhanced reusability. Optimum temperature remained unchanged, whereas optimum pH showed shift towards acidic range for immobilized enzyme. Increase in thermal stability of immobilized enzyme and non-toxic nature of functionalized graphene can be exploited for production of maltose in food and pharmaceutical industries.

  3. QM/MM and classical molecular dynamics simulation of histidine-tagged peptide immobilization on nickel surface

    Energy Technology Data Exchange (ETDEWEB)

    Yang Zhenyu [State Key Laboratory of Nonlinear Mechanics (LNM), Institute of Mechanics, Chinese Academy of Sciences, Beijing 100080(China); Zhao Yapu [State Key Laboratory of Nonlinear Mechanics (LNM), Institute of Mechanics, Chinese Academy of Sciences, Beijing 100080 (China)]. E-mail: yzhao@lnm.imech.ac.cn

    2006-05-15

    The hybrid quantum mechanics (QM) and molecular mechanics (MM) method is employed to simulate the His-tagged peptide adsorption to ionized region of nickel surface. Based on the previous experiments, the peptide interaction with one Ni ion is considered. In the QM/MM calculation, the imidazoles on the side chain of the peptide and the metal ion with several neighboring water molecules are treated as QM part calculated by 'GAMESS', and the rest atoms are treated as MM part calculated by 'TINKER'. The integrated molecular orbital/molecular mechanics (IMOMM) method is used to deal with the QM part with the transitional metal. By using the QM/MM method, we optimize the structure of the synthetic peptide chelating with a Ni ion. Different chelate structures are considered. The geometry parameters of the QM subsystem we obtained by QM/MM calculation are consistent with the available experimental results. We also perform a classical molecular dynamics (MD) simulation with the experimental parameters for the synthetic peptide adsorption on a neutral Ni(1 0 0) surface. We find that half of the His-tags are almost parallel with the substrate, which enhance the binding strength. Peeling of the peptide from the Ni substrate is simulated in the aqueous solvent and in vacuum, respectively. The critical peeling forces in the two environments are obtained. The results show that the imidazole rings are attached to the substrate more tightly than other bases in this peptide.

  4. A multi-defense strategy: Enhancing bactericidal activity of a medical grade polymer with a nitric oxide donor and surface-immobilized quaternary ammonium compound.

    Science.gov (United States)

    Pant, Jitendra; Gao, Jing; Goudie, Marcus J; Hopkins, Sean P; Locklin, Jason; Handa, Hitesh

    2017-08-01

    Although the use of biomedical devices in hospital-based care is inevitable, unfortunately, it is also one of the leading causes of the nosocomial infections, and thus demands development of novel antimicrobial materials for medical device fabrication. In the current study, a multi-defense mechanism against Gram-positive and Gram-negative bacteria is demonstrated by combining a nitric oxide (NO) releasing agent with a quaternary ammonium antimicrobial that can be covalently grafted to medical devices. Antibacterial polymeric composites were fabricated by incorporating an NO donor, S-nitroso-N-acetyl-penicillamine (SNAP) in CarboSil® polymer and top coated with surface immobilized benzophenone based quaternary ammonium antimicrobial (BPAM) small molecule. The results suggest that SNAP and BPAM individually have a different degree of toxicity towards Gram-positive and Gram-negative bacteria, while the SNAP-BPAM combination is effective in reducing both types of adhered viable bacteria equally well. SNAP-BPAM combinations reduced the adhered viable Pseudomonas aeruginosa by 99.0% and Staphylococcus aureus by 99.98% as compared to the control CarboSil films. Agar diffusion tests demonstrate that the diffusive nature of NO kills bacteria beyond the direct point of contact which the non-leaching BPAM cannot achieve alone. This is important for potential application in biofilm eradication. The live-dead bacteria staining shows that the SNAP-BPAM combination has more attached dead bacteria (than live) as compared to the controls. The SNAP-BPAM films have increased hydrophilicity and higher NO flux as compared to the SNAP films useful for preventing blood protein and bacterial adhesion. Overall the combination of SNAP and BPAM imparts different attributes to the polymeric composite that can be used in the fabrication of antimicrobial surfaces for various medical device applications. A significant increase in the biomedical device related infections (BDRIs), inability of

  5. Surface Immobilized His-tagged Azurin as a Model Interface for the Investigation of Vectorial Electron Transfer in Biological Systems

    International Nuclear Information System (INIS)

    Casalini, Stefano; Berto, Marcello; Kovtun, Alessandro; Operamolla, Alessandra; Di Rocco, Giulia; Facci, Paolo; Liscio, Andrea; Farinola, Gianluca M.; Borsari, Marco; Bortolotti, Carlo A.

    2015-01-01

    A model system for the electrochemical investigation of vectorial electron transfer in biological systems was designed, assembled and characterized. Gold electrodes, functionalized with a -OCH 3 terminated, aromatic self-assembled monolayer, were used as a substrate for the adsorption of variants of copper-containing, redox metalloprotein azurin. The engineered azurin bears a polyhistidine tag at its C-terminus. Thanks to the presence of the solvent exposed tag, which chelates Cu 2+ ions in solution, we introduced an exogenous redox centre. The different reduction potentials of the two redox centres and their positioning with respect to the surface are such that electron transfer from the exogenous copper centre and the electrode is mediated by the native azurin active site, closely paralleling electron transfer processes in naturally occurring multicentre metalloproteins.

  6. 100KE/KW fuel storage basin surface volumetric factors

    International Nuclear Information System (INIS)

    Conn, K.R.

    1996-01-01

    This Supporting Document presents calculations of surface Volumetric factors for the 100KE and 100KW Fuel Storage Basins. These factors relate water level changes to basin loss or additions of water, or the equivalent water displacement volumes of objects added to or removed from the basin

  7. Quantitative structure-retention relationship studies using immobilized artificial membrane chromatography I: amended linear solvation energy relationships with the introduction of a molecular electronic factor.

    Science.gov (United States)

    Li, Jie; Sun, Jin; Cui, Shengmiao; He, Zhonggui

    2006-11-03

    Linear solvation energy relationships (LSERs) amended by the introduction of a molecular electronic factor were employed to establish quantitative structure-retention relationships using immobilized artificial membrane (IAM) chromatography, in particular ionizable solutes. The chromatographic indices, log k(IAM), were determined by HPLC on an IAM.PC.DD2 column for 53 structurally diverse compounds, including neutral, acidic and basic compounds. Unlike neutral compounds, the IAM chromatographic retention of ionizable compounds was affected by their molecular charge state. When the mean net charge per molecule (delta) was introduced into the amended LSER as the sixth variable, the LSER regression coefficient was significantly improved for the test set including ionizable solutes. The delta coefficients of acidic and basic compounds were quite different indicating that the molecular electronic factor had a markedly different impact on the retention of acidic and basic compounds on IAM column. Ionization of acidic compounds containing a carboxylic group tended to impair their retention on IAM, while the ionization of basic compounds did not have such a marked effect. In addition, the extra-interaction with the polar head of phospholipids might cause a certain change in the retention of basic compounds. A comparison of calculated and experimental retention indices suggested that the semi-empirical LSER amended by the addition of a molecular electronic factor was able to reproduce adequately the experimental retention factors of the structurally diverse solutes investigated.

  8. Covalent immobilization of Pseudomonas cepacia lipase on semiconducting materials

    Energy Technology Data Exchange (ETDEWEB)

    Fernandez, Renny Edwin [Microelectronics and MEMS Laboratory, Department of Electrical Engineering, Indian Institute of Technology Madras, Chennai (India)], E-mail: rennyedwin@gmail.com; Bhattacharya, Enakshi [Microelectronics and MEMS Laboratory, Department of Electrical Engineering, Indian Institute of Technology Madras, Chennai (India)], E-mail: enakshi@ee.iitm.ac.in; Chadha, Anju [Department of Biotechnology, National Centre for Catalysis Research, Indian Institute of Technology Madras, Chennai (India)], E-mail: anjuc@iitm.ac.in

    2008-05-30

    Lipase from Pseudomonas cepacia was covalently immobilized on crystalline silicon, porous silicon and silicon nitride surfaces. The various stages of immobilization were characterized using FTIR (Fourier transform infrared) spectroscopy. The surface topography of the enzyme immobilized surfaces was investigated using scanning electron microscopy (SEM). The quantity of the immobilized active enzyme was estimated by the para-nitrophenyl palmitate (pNPP) assay. The immobilized lipase was used for triglyceride hydrolysis and the acid produced was detected by a pH sensitive silicon nitride surface as a shift in the C-V (capacitance-voltage) characteristics of an electrolyte-insulator-semiconductor capacitor (EISCAP) thus validating the immobilization method for use as a biosensor.

  9. Using shallow seismic tomography to characterize patterns of near-surface weathering and the mobile-immobile regolith transition: Implications for the erodibility and morphology of hillslopes.

    Science.gov (United States)

    Clarke, B. A.; Kirby, E.; Burbank, D. W.; West, N.

    2014-12-01

    We use 2D tomography of P- and S-wave velocities (Vp, Vs), based on seismic refraction and surface wave analyses, to characterize subsurface architecture and erodibility of hillslopes. Calibrating the seismic imagery with direct field observations allows us to quantify mechanical properties, image depth-dependent variations in weathering intensity, and identify the mobile-immobile regolith transition and differences in transport efficiency of mobile layers. We conducted a cross-CZO comparison of N- and S-facing slopes at Boulder Creek and Shale Hills CZOs (BcCZO and SSHCZO) to investigate how near-surface weathering and hillslope morphology are influenced by differences in regional geology and climatic as well as local variations in aspect-controlled microclimate. Niwot Ridge (BcCZO) is a high alpine site with minimal soil/veg cover, characterized by steeper S-facing hillslopes; whereas, SSHCZO is a temperate, densely-forested, soil-mantled site with steeper N-facing slopes. On Niwot Ridge, the depth of the weathering front and thickness of mobile regolith are substantially greater on shallower N-facing slopes; however, velocity-based estimates of transport efficiency are higher on S-facing slopes. Although, thin mobile regolith on S-facing slopes may be weaker (slower V), the lower gradient of N-facing slopes and southward asymmetry of the ridge divide, suggests greater transport efficiency on N-facing aspects. This can be explained by the dominance of frost/freeze process on N-facing slopes, which can efficiently develop and transport the thick mobile regolith. At SSHCZO, depths of weathering fronts are invariant with slope aspect, suggesting that aspect control is not a predominant mechanism driving regolith production. Mobile regolith thickness, however, is more than 2-fold greater on N-facing slopes. Additionally, mobile regolith on both slope aspects is primarily composed of well-developed soils. N-facing soils are thicker with greater cohesion, moisture, and

  10. Novel functionalized fluorescent polymeric nanoparticles for immobilization of biomolecules

    Science.gov (United States)

    Jain, Swati; Chattopadhyay, Sruti; Jackeray, Richa; Abid, C. K. V. Zainul; Singh, Harpal

    2013-07-01

    Novel, size controlled fluorescent polymeric nanoparticles (FPNP) were synthesized having acetoacetoxy functionality on the surface for immobilization of biomolecules which can be utilized as biomarkers and labels in fluoroimmunoassays. Core-shell nanoparticles of poly(styrene, St-methyl methacrylate, MMA-acetoacetoxy ethyl methacrylate, AAEM), stabilized by various concentrations of surfactant, sodium lauryl sulphate (SLS), were obtained by facile miniemulsion co-polymerization encapsulated with pyrene molecules in their hydrophobic core. Analytical, spectroscopic and imaging characterization techniques revealed the formation of stable, monodisperse, spherical nano sized particles exhibiting high luminescence properties. Particles with 1% SLS (S1) showed good dispersion stability and fluorescence intensity and were chosen as ideal candidates for further immobilization studies. Steady state fluorescence studies showed 10 times higher fluorescence intensity of S1 nanoparticles than that of pyrene solution in solvent-toluene at the same concentration. Environmental factors such as pH, ionic strength and time were found to have no effect on fluorescence intensity of FPNPs. Surface β-di-ketone groups were utilized for the covalent immobilization of enzyme conjugated antibodies without any activation or pre-treatment of nanoparticles.Novel, size controlled fluorescent polymeric nanoparticles (FPNP) were synthesized having acetoacetoxy functionality on the surface for immobilization of biomolecules which can be utilized as biomarkers and labels in fluoroimmunoassays. Core-shell nanoparticles of poly(styrene, St-methyl methacrylate, MMA-acetoacetoxy ethyl methacrylate, AAEM), stabilized by various concentrations of surfactant, sodium lauryl sulphate (SLS), were obtained by facile miniemulsion co-polymerization encapsulated with pyrene molecules in their hydrophobic core. Analytical, spectroscopic and imaging characterization techniques revealed the formation of stable

  11. Optimizing immobilized enzyme performance in cell-free environments to produce liquid fuels.

    Energy Technology Data Exchange (ETDEWEB)

    Kumar, Sanat

    2015-02-05

    The overall goal of this project was to optimize enzyme performance for the production of bio-diesel fuel. Enzyme immobilization has attracted much attention as a means to increase productivity. Mesorporous silica materials have been known to be best suited for immobilizing enzymes. A major challenge is to ensure that the enzymatic activity is retained after immobilization. Two major factors which drive enzymatic deactivation are protein-surface and inter-protein interactions. Previously, we studied protein stability inside pores and how to optimize protein-surface interactions to minimize protein denaturation. In this work we studied eh effect of surface curvature and chemistry on inter-protein interactions. Our goal was to find suitable immobilization supports which minimize these inter-protein interactions. Our studies carried out in the frame work of Hydrophobic-Polar (HP) model showed that enzymes immobilized inside hydrophobic pores of optimal sizes are best suited to minimize these inter-protein interactions. Besides, this study is also of biological importance to understand the role of chaperonins in protein disaggregation. Both of these aspects profited immensely with collaborations with our experimental colleague, Prof. Georges Belfort (RPI), who performed the experimental analog of our theoretical works.

  12. Surface water quality assessment using factor analysis | Boyacioglu ...

    African Journals Online (AJOL)

    In this study, the factor analysis technique is applied to surface water quality data sets obtained from the Buyuk Menderes River Basin, Turkey, during two different hydrological periods. Results show that the indices which changed the quality of water in two seasons and locations differed. During low-flow conditions, water ...

  13. Studies on Prevalence and Risk Factors for Hepatitis B Surface ...

    African Journals Online (AJOL)

    Studies on Prevalence and Risk Factors for Hepatitis B Surface Antigen among Secondary School Students in North-central, Nigeria. ... high among the studied population. This suggests that public awareness on the virus be accorded urgent attention, while vaccination programme should be improved in the community.

  14. Immobilization of Mitochondria on Graphene

    Science.gov (United States)

    2013-08-29

    poly- acryloylaminopropanol (AAP) and poly-vinyl alcohol (PVA) reduces the adsorption of mitochondria on the surface (Whiting, C.E., and A. Edgar...poly-L-lysine has also been reported for immobilization of yeast mitochondria. Coating was performed by repetitive washing of cover slips with 0.02...of Poly-L-lysine Applications of PLL PLL is a production of bacterial fermentation and is used as a food preservative. In biology, PLL is used in

  15. Upregulation of interleukin-1β/transforming growth factor-β1 and hypoxia relate to molecular mechanisms underlying immobilization-induced muscle contracture.

    Science.gov (United States)

    Honda, Yuichiro; Sakamoto, Junya; Nakano, Jiro; Kataoka, Hideki; Sasabe, Ryo; Goto, Kyo; Tanaka, Miho; Origuchi, Tomoki; Yoshimura, Toshiro; Okita, Minoru

    2015-09-01

    In this study we investigated the molecular mechanism underlying muscle contracture in rats. The rats were divided into immobilization and control groups, and soleus muscles of the right and left sides were selected for analyses. The levels of CD11b and α-SMA protein, IL-1β, and TGF-β1 mRNA, and type I and III collagen protein and mRNA were significantly greater in the immobilization group than in the control group at all time-points. HIF-1α mRNA levels were significantly higher in the immobilization group at 4 weeks. Moreover, HIF-1α, α-SMA, and type I collagen levels were significantly higher at 4 weeks than at 1 and 2 weeks in the immobilization group. In the early stages of immobilization, upregulation of IL-1β/TGF-β1 via macrophages may promote fibroblast differentiation that could affect muscle contracture. The soleus muscle became hypoxic in the later stages of immobilization, suggesting that hypoxia influences the progression of muscle contracture. © 2014 Wiley Periodicals, Inc.

  16. Essential role of structural integrity and firm attachment of surface-anchored epidermal growth factor in adherent culture of neural stem cells.

    Science.gov (United States)

    Nakaji-Hirabayashi, Tadashi; Kato, Koichi; Iwata, Hiroo

    2008-11-01

    Surface immobilization of proteins provides various biomaterials that permit the control of cellular functions through protein-protein interactions. Our previous study demonstrated that human epidermal growth factor carrying a hexahistidine sequence at the C-terminus (hEGF-His) could be anchored to the Ni-chelated surface by coordination, providing the versatile substrate for the selective proliferation of neural stem cells. The present study was undertaken to gain deeper insights into the basis for such an outstanding property of the surface with coordinated hEGF-His. For this purpose, the structure of the coordinated hEGF-His was analyzed by multiple internal reflection-infrared absorption spectroscopy. In addition, stability of coordinate bonds was assessed under cell culture conditions using a spatially-restricted anchoring technique. These data were compared to the results obtained from surfaces with covalently immobilized and physically adsorbed hEGF-His. The results presented here demonstrate that coordinated hEGF-His remains its intact conformation and is firmly anchored to the surface during cell culture. These attributes are both crucial for establishing the adherent culture and hence selective expansion of neural stem cells.

  17. Application of magnetic nanoparticles in smart enzyme immobilization.

    Science.gov (United States)

    Vaghari, Hamideh; Jafarizadeh-Malmiri, Hoda; Mohammadlou, Mojgan; Berenjian, Aydin; Anarjan, Navideh; Jafari, Nahideh; Nasiri, Shahin

    2016-02-01

    Immobilization of enzymes enhances their properties for efficient utilization in industrial processes. Magnetic nanoparticles, due to their high surface area, large surface-to-volume ratio and easy separation under external magnetic fields, are highly valued. Significant progress has been made to develop new catalytic systems that are immobilized onto magnetic nanocarriers. This review provides an overview of recent developments in enzyme immobilization and stabilization protocols using this technology. The current applications of immobilized enzymes based on magnetic nanoparticles are summarized and future growth prospects are discussed. Recommendations are also given for areas of future research.

  18. Anti-inflammatory and antioxidant effect of cerium dioxide nanoparticles immobilized on the surface of silica nanoparticles in rat experimental pneumonia.

    Science.gov (United States)

    Serebrovska, Z; Swanson, R J; Portnichenko, V; Shysh, A; Pavlovich, S; Tumanovska, L; Dorovskych, A; Lysenko, V; Tertykh, V; Bolbukh, Y; Dosenko, V

    2017-08-01

    A massage with the potent counter-inflammatory material, cerium dioxide nanoparticles, is promising and the antioxidant properties of CeO 2 are considered the main, if not the only, mechanism of this action. Nevertheless, the elimination of ceria nano-particles from the organism is very slow and there is a strong concern for toxic effect of ceria due to its accumulation. To overcome this problem, we engineered a combined material in which cerium nanoparticles were immobilized on the surface of silica nanoparticles (CeO 2 NP), which is shown to be easily removed from an organism and could be used as carriers for nano-ceria. In our study particle size was 220±5nm, Zeta-potential -4.5mV (in water), surface charge density -17.22μC/cm 2 (at pH 7). Thirty-six male Wistar rats, 5 months old and 250-290g were divided into four groups: 1) control; 2) CeO 2 NP treatment; 3) experimental pneumonia (i/p LPS injection, 1mg/kg); and 4) experimental pneumonia treated with CeO 2 NP (4 times during the study in dosage of 0.6mg/kg with an orogastric catheter). Gas exchange and pulmonary ventilation were measured four times: 0, 1, 3 and 24h after LPS injection in both untreated and CeO 2 NP-treated animals. The mRNA of TNF-α, Il-6, and CxCL2 were determined by RT-PCR. ROS-generation in blood plasma and lung tissue homogenates were measured by means of lucigenin- and luminol-enhanced chemiluminescence. Endotoxemia in the acute phase was associated with: (1) pathological changes in lung morphology; (2) increase of ROS generation; (3) enhanced expression of CxCL2; and (4) a gradual decrease of VO 2 and V E . CeO2 NP treatment of intact animals did not make any changes in all studied parameters except for a significant augmentation of VO 2 and V E. CeO 2 NP treatment of rats with pneumonia created positive changes in diminishing lung tissue injury, decreasing ROS generation in blood and lung tissue and decreasing pro-inflammatory cytokine expression (TNF-α, Il-6 and CxCL2). Oxygen

  19. Topography measurements for determining the decay factors in surface replication

    International Nuclear Information System (INIS)

    Song, J; Zheng, A; Vorburger, T V; Rubert, P

    2008-01-01

    The electro-forming technique is used at National Institute of Standards and Technology (NIST) for the production of standard reference material (SRM) 2461 standard casings to support nationwide ballistics measurement traceability and measurement quality control in the US. In order to ensure that the SRM casings are produced with virtually the same surface topography, it is necessary to test the decay factors of the replication process. Twenty-six replica casings are replicated from the same master casing for the decay factor tests. The NIST topography measurement system is used for measurements and correlations of surface topography. The topography decays are quantified by the cross-correlation function maximum CCF max . Based on the test, it is expected that 256 SRM casings can be replicated from the same master with CCF max values higher than 95%

  20. Development of an immunosensor using oriented immobilized anti-OmpW for sensitive detection of Vibrio cholerae by surface plasmon resonance.

    Science.gov (United States)

    Taheri, Ramezan Ali; Rezayan, Ali Hossein; Rahimi, Fereshteh; Mohammadnejad, Javad; Kamali, Mehdi

    2016-12-15

    The first SPR sensor for detection of bacteria was reported in 1998 with high detection limit as much as 10(7)cfu/mL. Since then, a lot of effort has been made to lower detection limit and increase sensitivity of detection mainly by using of different assay formats, immobilization strategies, suitable antibodies, minimizing non-specific adsorption and improving the quality of SPR devices. The aim of this paper is to introduce the potential of an antibody against recombinant outer membrane protein (anti-OmpW) in sensitive detection of Vibrio cholerae by developing an immunosensor based on SPR and compare the sensitivity of this method with former report for detection of V. cholerae published in 2006. Recombinant OmpW antigen (a bacterial outer-membrane protein) of V. cholerae was expressed and purified and raising of polyclonal rabbit anti-OmpW was done. Protein G was covalently immobilized on 11-MUA SAM via amine coupling and bioaffinity-based oriented immobilization of anti-OmpW was done on protein G layer. The results showed high affinity interaction between OmpW and anti-OmpW (KD=2.4×10(-9)M) and the detection limit of fabricated immunosensor was 43 cells/mL. The apparent reasons for achieving this low LOD are discussed. Copyright © 2016 Elsevier B.V. All rights reserved.

  1. Plutonium immobilization -- Can loading

    International Nuclear Information System (INIS)

    Kriikku, E.

    2000-01-01

    The Savannah River Site (SRS) will immobilize excess plutonium in the proposed Plutonium Immobilization Project (PIP). The PIP adds the excess plutonium to ceramic pucks, loads the pucks into cans, and places the cans into DWPF canisters. This paper discusses the PIP process steps, the can loading conceptual design, can loading equipment design, and can loading work completed

  2. Immobilized waste leaching

    International Nuclear Information System (INIS)

    Suarez, A.A.

    1989-01-01

    The main mechanism by which the immobilized radioactive materials can return to biosphere is the leaching due to the intrusion of water into the repositories. Some mathematical models and experiments utilized to evaluate the leaching rates in different immobilization matrices are described. (author) [pt

  3. Factors Predicting the Ocular Surface Response to Desiccating Environmental Stress

    Science.gov (United States)

    Alex, Anastasia; Edwards, Austin; Hays, J. Daniel; Kerkstra, Michelle; Shih, Amanda; de Paiva, Cintia S.; Pflugfelder, Stephen C.

    2013-01-01

    Purpose. To identify factors predicting the ocular surface response to experimental desiccating stress. Methods. The ocular surfaces of both eyes of 15 normal and 10 dry eye subjects wearing goggles were exposed to a controlled desiccating environment (15%–25% relative humidity and 2–5 L/min airflow) for 90 minutes. Eye irritation symptoms, blink rate, tear meniscus dimensions, noninvasive (RBUT) and invasive tear break-up time, and corneal fluorescein and conjunctival lissamine green-dye staining were recorded before and after desiccating stress. Pre- and postexposure measurements were compared, and Pearson correlations between clinical parameters before and after desiccating stress were calculated. Results. Corneal and conjunctival dye staining significantly increased in all subjects following 90-minute exposure to desiccating environment, and the magnitude of change was similar in normal and dry eye subjects; except superior cornea staining was greater in dry eye. Irritation severity in the desiccating environment was associated with baseline dye staining, baseline tear meniscus height, and blink rate after 45 minutes. Desiccation-induced change in corneal fluorescein staining was inversely correlated to baseline tear meniscus width, whereas change in total ocular surface dye staining was inversely correlated to baseline dye staining, RBUT, and tear meniscus height and width. Blink rate from 30 to 90 minutes in desiccating environment was higher in the dry eye than normal group. Blink rate significantly correlated to baseline corneal fluorescein staining and environmental-induced change in corneal fluorescein staining. Conclusions. Ocular surface dye staining increases in response to desiccating stress. Baseline ocular surface dye staining, tear meniscus height, and blink rate predict severity of ocular surface dye staining following exposure to a desiccating environment. PMID:23572103

  4. Immobilization of inorganic pyrophosphatase on nanodiamond particles retaining its high enzymatic activity.

    Science.gov (United States)

    Rodina, Elena V; Valueva, Anastasiya V; Yakovlev, Ruslan Yu; Vorobyeva, Nataliya N; Kulakova, Inna I; Lisichkin, Georgy V; Leonidov, Nikolay B

    2015-12-21

    Nanodiamond (ND) particles are popular platforms for the immobilization of molecular species. In the present research, enzyme Escherichia coli inorganic pyrophosphatase (PPase) was immobilized on detonation ND through covalent or noncovalent bonding and its enzymatic activity was characterized. Factors affecting adsorption of PPase such as ND size and surface chemistry were studied. The obtained material is a submicron size association of ND particles and protein molecules in approximately equal amounts. Both covalently and noncovalently immobilized PPase retains a significant enzymatic activity (up to 95% of its soluble form) as well as thermostability. The obtained hybrid material has a very high enzyme loading capacity (∼1 mg mg(-1)) and may be considered as a promising delivery system of biologically active proteinaceous substances, particularly in the treatment of diseases such as calcium pyrophosphate crystal deposition disease and related pathologies. They can also be used as recoverable heterogeneous catalysts in the traditional uses of PPase.

  5. Calculation of Operations Efficiency Factors for Mars Surface Missions

    Science.gov (United States)

    Layback, Sharon L.

    2014-01-01

    For planning of Mars surface missions, to be operated on a sol-by-sol basis by a team on Earth (where a "sol" is a Martian day), activities are described in terms of "sol types" that are strung together to build a surface mission scenario. Some sol types require ground decisions based on a previous sol's results to feed into the activity planning ("ground in the loop"), while others do not. Due to the differences in duration between Earth days and Mars sols, for a given Mars local solar time, the corresponding Earth time "walks" relative to the corresponding times on the prior sol/day. In particular, even if a communication window has a fixed Mars local solar time, the Earth time for that window will be approximately 40 minutes later each succeeding day. Further complexity is added for non-Mars synchronous communication relay assets, and when there are multiple control centers in different Earth time zones. The solution is the development of "ops efficiency factors" that reflect the efficiency of a given operations configuration (how many and location of control centers, types of communication windows, synchronous or non-synchronous nature of relay assets, sol types, more-or-less sustainable operations schedule choices) against a theoretical "optimal" operations configuration for the mission being studied. These factors are then incorporated into scenario models in order to determine the surface duration (and therefore minimum spacecraft surface lifetime) required to fulfill scenario objectives. The resulting model is used to perform "what-if" analyses for variations in scenario objectives. The ops efficiency factor is the ratio of the figure of merit for a given operations factor to the figure of merit for the theoretical optimal configuration. The current implementation is a pair of models in Excel. The first represents a ground operations schedule for 500 sols in each operations configuration for the mission being studied (500 sols was chosen as being a long

  6. A disposal centre for immobilized nuclear waste

    International Nuclear Information System (INIS)

    1980-02-01

    This report describes a conceptual design of a disposal centre for immobilized nuclear waste. The surface facilities consist of plants for the preparation of steel cylinders containing nuclear waste immobilized in glass, shaft headframe buildings and all necessary support facilities. The underground disposal vault is located on one level at a depth of 1000 m. The waste cylinders are emplaced into boreholes in the tunnel floors. All surface and subsurface facilities are described, operations and schedules are summarized, and cost estimates and manpower requirements are given. (auth)

  7. Plutonium Disposition by Immobilization

    International Nuclear Information System (INIS)

    Gould, T.; DiSabatino, A.; Mitchell, M.

    2000-01-01

    The ultimate goal of the Department of Energy (DOE) Immobilization Project is to develop, construct, and operate facilities that will immobilize between 17 to 50 tonnes (MT) of U.S. surplus weapons-usable plutonium materials in waste forms that meet the ''spent fuel'' standard and are acceptable for disposal in a geologic repository. Using the ceramic can-in-canister technology selected for immobilization, surplus plutonium materials will be chemically combined into ceramic forms which will be encapsulated within large canisters of high level waste (HLW) glass. Deployment of the immobilization capability should occur by 2008 and be completed within 10 years. In support of this goal, the DOE Office of Fissile Materials Disposition (MD) is conducting development and testing (D and T) activities at four DOE laboratories under the technical leadership of Lawrence Livermore National Laboratory (LLNL). The Savannah River Site has been selected as the site for the planned Plutonium Immobilization Plant (PIP). The D and T effort, now in its third year, will establish the technical bases for the design, construction, and operation of the U. S. capability to immobilize surplus plutonium in a suitable and cost-effective manner. Based on the D and T effort and on the development of a conceptual design of the PIP, automation is expected to play a key role in the design and operation of the Immobilization Plant. Automation and remote handling are needed to achieve required dose reduction and to enhance operational efficiency

  8. BMP-2 immobilized PLGA/hydroxyapatite fibrous scaffold via polydopamine stimulates osteoblast growth.

    Science.gov (United States)

    Zhao, Xingyu; Han, Yu; Li, Jiawei; Cai, Bo; Gao, Hang; Feng, Wei; Li, Shuqiang; Liu, Jianguo; Li, Dongsong

    2017-09-01

    Combining biomaterials scaffolds with bone morphogenetic protein-2 (BMP-2) is currently used to promote the regeneration of bone tissue. However, the traditional strategies used to add BMP-2 into the polymer scaffolds directly suffer from limitations that can result in lower growth factor loading and damage the bioactivity of growth factors. In this study, we report the fabrication of poly(lactide-co-glycolide)/hydroxyapatite (PLGA/HA) composite fibrous scaffolds via melt-spinning method to mimic native extracellular matrix (ECM). In order to effectively immobilize BMP-2 on PLGA/HA composite fibrous scaffolds, the surface of the scaffold was modified with polydopamine (PDA) (PDA-PLGA/HA). PDA was chosen as an adhesive polymeric bridge-layer between PLGA/HA fibrous scaffolds and BMP-2. Analysis of the scaffold using scanning electron microscopy, Fourier transform infrared spectroscopy and X-ray photoelectron spectroscope revealed that the PDA coating was attached to the scaffold surface. Moreover, analysis of the scaffold using water contact angle demonstrated an increased hydrophilicity via PDA modification. Furthermore, the PDA coating effectively immobilized BMP-2 on the PDA-PLGA/HA fibrous scaffold and a sustained release profile of BMP-2 was achieved in the BMP-2-immobilized PLGA/HA fibrous scaffold. In vitro experiments showed that BMP-2-immobilized PLGA/HA fibrous scaffold significantly promoted the attachment and proliferation of MC3T3-E1 cells. More importantly, the ALP activity, mRNA expression of osteosis-related genes and calcium deposition in MC3T3-E1 cells cultured on BMP-2-immobilized PLGA/HA fibrous scaffold were significantly increased. These results collectively demonstrate that the BMP-2-immobilized PLGA/HA fibrous scaffold is a promising candidate for bone regeneration. Copyright © 2017 Elsevier B.V. All rights reserved.

  9. Preparation of immobilized growing cells and enzymatic hydrolysis of sawdust

    International Nuclear Information System (INIS)

    Kumakura, M.; Kaetsu, I.

    1984-01-01

    Trichoderma reesei cells were immobilized by radiation polymerization using porous materials such as non-woven material and sawdust, and the enzymatic hydrolysis of sawdust with the enzyme solution from the immobilized growing cells was studied. The filter paper activity, which shows the magnitude of cellulase production in the immobilized cells, was comparable with that in the intact cells. The filter paper activity was affected by addition concentration of monomer and porous materials. The cells in the immobilized cells grew to be adhered on the surface of the fibrous polymers. Sawdust, which was pretreated by irradiation technique, was effectively hydrolyzed with the enzyme solution resulting from the culture of the immobilized cells, in which the glucose yield increased increasing the culture time of the immobilized cells. (author)

  10. Isomaltulose production using free and immobilized Serratia ...

    African Journals Online (AJOL)

    André

    2016-05-18

    May 18, 2016 ... Isomaltulose is a low cariogenic sweetener used as a substitute for sucrose in the food industry. In this study, isomaltulose production by Serratia plymuthica ATCC 15928 was performed using free and immobilized cells. Response Surface Methodology was employed to evaluate the influence of.

  11. Isomaltulose production using free and immobilized Serratia ...

    African Journals Online (AJOL)

    Isomaltulose is a low cariogenic sweetener used as a substitute for sucrose in the food industry. In this study, isomaltulose production by Serratia plymuthica ATCC 15928 was performed using free and immobilized cells. Response Surface Methodology was employed to evaluate the influence of temperature, wet cell mass ...

  12. Enzyme production in immobilized Trichoderma reesei cells with hydrophobic polymers prepared by radiation polymerization method

    International Nuclear Information System (INIS)

    Luzhao Xin; Kumakura, Minoru; Kaetsu, Isao

    1993-01-01

    Trichoderma reesei cells were immobilized on paper covered with hydrophobic monomer, trimethylpropane triacrylate by radiation polymerization. The effect of immobilization condition on enzyme productivity was studied by measuring filter paper and cellobiose activity. The cells were adhered and grew on the surface of the carrier with the polymer giving high enzyme productivity in the immobilized cells in comparison with the free cells. Optimum concentration and volume of the coating monomer for the preparation of the immobilized cells were obtained. (author)

  13. Increase in stability of cellulase immobilized on functionalized magnetic nanospheres

    International Nuclear Information System (INIS)

    Zhang, Wenjuan; Qiu, Jianhui; Feng, Huixia; Zang, Limin; Sakai, Eiichi

    2015-01-01

    Functionalized magnetic nanospheres were prepared by co-condensation of tetraethylorthosilicate with three different amino-silanes: 3-(2-aminoethylamino propyl)-triethoxysilane (AEAPTES), 3-(2-aminoethylamino propyl)-trimethoxysilane (AEAPTMES) and 3-aminopropyltriethoxysilane (APTES). Then three functionalized magnetic nanospheres were used as supports for immobilization of cellulase. The three functionalized magnetic nanospheres with core–shell morphologies exhibited higher capacity for cellulase immobilization than unfunctionalized magnetic nanospheres. The increasing of surface charge of functionalized magnetic nanospheres leads to an enhancement of the capacity of cellulase immobilization. Particularly, AEAPTMES with methoxy groups was favored to be hydrolyzed and grafted on unfunctionalized magnetic nanospheres than the others. AEAPTMES functionalized magnetic nanospheres with the highest zeta potential (29 mV) exhibited 87% activity recovery and the maximum amount of immobilized cellulase was 112 mg/g support at concentration of initial cellulase of 8 mg/mL. Immobilized cellulase on AEAPTMES functionalized magnetic nanospheres had higher temperature stability and broader pH stability than other immobilized cellulases and free cellulase. In particular, it can be used in about 40 °C, demonstrating the potential of biofuel production using this immobilized cellulase. - Highlights: • Three Amino-silane modified magnetic nanospheres were prepared. • Cellulase immobilized AEAPTMES functionalized magnetic nanospheres had higher temperature stability and broader pH stability than free cellulase. • The potential of biofuel production using this immobilized cellulase

  14. Covalent immobilization of invertase on PAMAM-dendrimer modified superparamagnetic iron oxide nanoparticles

    International Nuclear Information System (INIS)

    Uzun, K.; Cevik, E.; Senel, M.; Soezeri, H.; Baykal, A.; Abasiyanik, M. F.; Toprak, M. S.

    2010-01-01

    In this study, polyamidoamine (PAMAM) dendrimer was synthesized on the surface of superparamagnetite nanoparticles to enhance invertase immobilization. The amount of immobilized enzyme on the surface-hyperbranched magnetite nanoparticle was up to 2.5 times (i.e., 250%) as much as that of magnetite nanoparticle modified with only amino silane. Maximum reaction rate (V max ) and Michaelis-Menten constant (K m ) were determined for the free and immobilized enzymes. Various characteristics of immobilized invertase such as; the temperature activity, thermal stability, operational stability, and storage stability were evaluated and results revealed that stability of the enzyme is improved upon immobilization.

  15. Laccase-polyacrylonitrile nanofibrous membrane: highly immobilized, stable, reusable, and efficacious for 2,4,6-trichlorophenol removal.

    Science.gov (United States)

    Xu, Ran; Chi, Chenglong; Li, Fengting; Zhang, Bingru

    2013-12-11

    Increasing attention has been given to nanobiocatalysis for commercial applications. In this study, laccase was immobilized on polyacrylonitrile (PAN) nanofibrous membranes through ethanol/HCl method of amidination reaction and successfully applied for removal of 2,4,6-trichlorophenol (TCP) from water. PAN membranes with fiber diameters from 200 nm to 300 nm were fabricated via electrospinning and provided a large surface area for enzyme immobilization and catalytic reactions. Images of scanning electron microscope demonstrated the enzyme molecules were aggregated on the nanofiber surface. The immobilized laccase exhibited 72% of the free enzyme activity and kept 60% of its initial activity after 10 operation cycles. Moreover, the storage stability of the immobilized laccase was considered excellent because they maintained more than 92% of the initial activity after 18 days of storage, whereas the free laccase retained only 20%. The laccase-PAN nanofibrous membranes exhibited high removal efficiency of TCP under the combined actions of biodegradation and adsorption. More than 85% of the TCP was removed under optimum conditions. Effects of various factors on TCP removal efficiency of the immobilized laccase were analyzed. Results suggest that laccase-PAN nanofibrous membranes can be used in removing TCP from aqueous sources and have potential for use in other commercial applications.

  16. Haloalkane hydrolysis with an immobilized haloalkane dehalogenase.

    Science.gov (United States)

    Dravis, B C; Swanson, P E; Russell, A J

    2001-11-20

    Haloalkane dehalogenase from Rhodococcus rhodochrous was covalently immobilized onto a polyethyleneimine impregnated gamma-alumina support. The dehalogenating enzyme was found to retain greater than 40% of its original activity after immobilization, displaying an optimal loading (max. activity/supported protein) of 70 to 75 mg/g with an apparent maximum (max. protein/support) of 156 mg/g. The substrate, 1,2,3-trichloropropane, was found to favorably partition (adsorb) onto the inorganic alumina carrier (10 to 20 mg/g), thereby increasing the local reactant concentration with respect to the catalyst's environment, whereas the product, 2,3-dichloropropan-1-ol, demonstrated no affinity. Additionally, the inorganic alumina support exhibited no adverse effects because of solvent/component incompatibilities or deterioration due to pH variance (pH 7.0 to 10.5). As a result of the large surface area to volume ratio of the support matrix and the accessibility of the bound protein, the immobilized biocatalyst was not subject to internal mass transfer limitations. External diffusional restrictions could be eliminated with simple agitation (mixing speed: 50 rpm; flux: 4.22 cm/min). The pH-dependence of the immobilized dehalogenase was essentially the same as that for the native enzyme. Finally, both the thermostability and resistance toward inactivation by organic solvent were improved by more than an order of magnitude after immobilization. Copyright 2001 John Wiley & Sons, Inc.

  17. Optimization of Immobilization of Nanodiamonds on Graphene

    International Nuclear Information System (INIS)

    Pille, A; Lange, S; Utt, K; Eltermann, M

    2015-01-01

    We report using simple dip-coating method to cover the surface of graphene with nanodiamonds for future optical detection of defects on graphene. Most important part of the immobilization process is the pre-functionalization of both, nanodiamond and graphene surfaces to obtain the selectiveness of the method. This work focuses on an example of using electrostatic attraction to confine nanodiamonds to graphene. Raman spectroscopy, microluminescence imaging and scanning electron microscopy were applied to characterize obtained samples. (paper)

  18. [The immobilization syndrome].

    Science.gov (United States)

    Dénes, Z

    1996-08-11

    Prolonged inactivity, bed rest causes pathological changes in most organs and systems of the body generally known as immobilization syndrome. These changes became known in the past 50 years in addition to scientific experiences. The author reviews pathological changes that occur with inactivity, discusses the predisposition of the elderly to the complication of bed rest and makes recommendations for treatment and prevention. While the effects of immobilization are mostly reversible in young subjects, older persons have considerably more difficulty recovering, it can even lead to the loss of independence. Minimizing duration of bed rest, early ambulation, good nursing care and physiotherapy can prevent many of the complications of inactivity and bed rest.

  19. Immobilization of microorganisms. Part 1. Preparation of immobilized Lactobacillus bulgaricus

    Energy Technology Data Exchange (ETDEWEB)

    Lee, K.H.

    1981-01-01

    The immobilization of Lactobacillus bulgaricus on polyacrylamide and on alginate beads was investigated. The most active immobilized cells were obtained by entrapment in Ca alginate beads. These immobilized microbial cells, when introduced into 4.5% lactose solution and whey solution showed maximum relative activity of 28% for lactose and 18% for whey compared to free cells.

  20. Limb immobilization and corticobasal syndrome.

    Science.gov (United States)

    Graff-Radford, Jonathan; Boeve, Bradley F; Drubach, Daniel A; Knopman, David S; Ahlskog, J Eric; Golden, Erin C; Drubach, Dina I; Petersen, Ronald C; Josephs, Keith A

    2012-12-01

    Recently, we evaluated two patients with corticobasal syndrome (CBS) who reported symptom onset after limb immobilization. Our objective was to investigate the association between trauma, immobilization and CBS. The charts of forty-four consecutive CBS patients seen in the Mayo Clinic Alzheimer Disease Research Center were reviewed with attention to trauma and limb immobilization. 10 CBS patients (23%) had immobilization or trauma on the most affected limb preceding the onset or acceleration of symptoms. The median age at onset was 61. Six patients manifested their first symptoms after immobilization from surgery or fracture with one after leg trauma. Four patients had pre-existing symptoms of limb dysfunction but significantly worsened after immobilization or surgery. 23 percent of patients had immobilization or trauma of the affected limb. This might have implications for management of CBS, for avoiding injury, limiting immobilization and increasing movement in the affected limb. Copyright © 2012 Elsevier Ltd. All rights reserved.

  1. Laser-assisted immobilization of colloid silver nanoparticles on polyethyleneterephthalate

    Science.gov (United States)

    Siegel, Jakub; Lyutakov, Oleksiy; Polívková, Markéta; Staszek, Marek; Hubáček, Tomáš; Švorčík, Václav

    2017-10-01

    Immobilization of nanoobjects on the surface of underlying material belongs to current issues of material science. Such altered materials exhibits completely exceptional properties exploitable in a broad spectrum of industrially important applications ranging from catalysts up to health-care industry. Here we present unique approach for immobilization of electrochemically synthesized silver nanoparticles on polyethyleneterephthalate (PET) foil whose essence lies in physical incorporation of particles into thin polymer surface layer induced by polarized excimer laser light. Changes in chemical composition and surface structure of polymer after particle immobilization were recorded by wide range of analytical techniques such as ARXPS, EDX, RBS, AAS, Raman, ICP-MS, DLS, UV-vis, SEM, TEM, and AFM. Thorough analysis of both nanoparticles entering the immobilization step as well as modified PET surface allowed revealing the mechanism of immobilization process itself. Silver nanoparticles were physically embedded into a thin surface layer of polymer reaching several nanometers beneath the surface rather than chemically bonded to PET macromolecules. Laser-implanted nanoparticles open up new possibilities especially in the development of the next generation cell-conform antimicrobial coatings of polymeric materials, namely due to the considerable immobilization strength which is strong enough to prevent particle release into the surrounding environment.

  2. Structural and catalytic properties of immobilized α-amylase from Laceyella sacchari TSI-2.

    Science.gov (United States)

    Shukla, Rushit J; Singh, Satya P

    2016-04-01

    One of the approaches to address the issues of the cost of production, recovery and reusability of the extremozymes can be immobilization. In this report, we describe immobilization of an α-amylase from Laceyella sacchari TSI-2 and characterization of the immobilized enzyme. The enzyme was immobilized on 6 different matrices using entrapment, ionic binding and surface adsorption. The DEAE cellulose with glutaraldehyde crosslinking appeared most effective for the immobilization with high operational stability. While the temperature optima and thermal stability of the immobilized α-amylase shifted from 60 to 70°C with increased half-life, the pH optima remain unaltered while pH stability was shifted from 6 to 7. The stability of the immobilized enzyme improved in solvents. The enzyme catalysis in surfactants enhanced, while the Km and Vmax were reduced after immobilization. The structural features of the immobilized enzyme as probed by FT-IR established the role of aliphatic amines, esters and alkenes in immobilization. The starch hydrolysis efficiency of the immobilized enzyme was 15.55%. The immobilized enzyme in various detergents was highly efficient in removing the starch stain from cotton cloth. Taken together, the α-amylase turned more stable after immobilization and can be a favored choice for applications. Copyright © 2015 Elsevier B.V. All rights reserved.

  3. Factors influencing the surface quality of polished tool steels

    International Nuclear Information System (INIS)

    Rebeggiani, S; Rosén, B-G

    2014-01-01

    Today’s demands on surface quality of moulds for injection moulding of plastic components involve no/low defect contents and roughness levels in the nm-range for high gloss applications. Material properties as well as operating conditions influence the mould finish, and thus the final surface of moulded products. This paper focuses on how particle content and different polishing strategies influence final surface qualities of moulds. Visual estimations of polished tool steel samples were combined with non-contact 3D-surface texture analysis in order to correlate traditional assessments to more quantitative methods, and to be able to analyse the surfaces at nanometre-level. It was found that steels with a lower proportion of particles, like carbides and oxides, gave rise to smoother polished surfaces. In a comparative study of polishers from different polishing shops, it was found that while different surface preparation strategies can lead to similar final roughness, similar preparation techniques can produce high-quality surfaces from different steel grades. However, the non-contact 3D-surface texture analysis showed that not all smooth polished surfaces have desirable functional topographies for injection moulding of glossy plastic components. (paper)

  4. Study of the surface crystallization and resistance to dissolution of niobium phosphate glasses for nuclear waste immobilization; Estudo da cristalizacao superficial e da resistencia a dissolucao de vidros niobofosfatos visando a imobilizacao de rejeitos radioativos

    Energy Technology Data Exchange (ETDEWEB)

    Vieira, Heveline

    2008-07-01

    The surface crystallization and the dissolution rate of three phosphate glass compositions containing different amounts of niobium oxide were studied. The glasses were named Nb30, Nb37, and Nb44 according to the nominal content of niobium oxide in the glass composition. The three compositions were evaluated keeping the P{sub 2}O{sub 5}/K{sub 2}O ratio constant and varying the amount of Nb{sub 2}O{sub 5}. These glasses were produced by melting appropriate chemical compounds at 1500 deg C for 0.5 hour. The crystalline phases which were nucleated on the glass surface after heat treatment were determined by X-ray diffraction. The crystalline structures depend on the amount of niobium oxide in the glass composition. The crystal morphologies were observed by using an optical microscope, and their characteristics are specific for each kind of crystalline phase. The crystal growth rate and the surface nuclei density were determined for each glass composition, and they depend on each crystalline phase nucleated on the surface. From the differential thermal analysis curves it was determined that the Nb44 glass containing 46.5 mol por cent of niobium oxide is the most thermally stable against crystallization when compared to the Nb30 and Nb37 glasses. According to the activation energies determined for crystal growth on the surface of each glass type, the Nb44 glass can also be considered the most resistant one against crystallization. The dissolution rate for the Nb44 glass after 14 days immersed in an aqueous solution with pH equals to 7 at 90 deg C is the lowest (9.0 x 10{sup -7} g. cm{sup -2} . day{sup -1}) when compared to the other two glass compositions. The dissolution rates in acidic and neutral solutions of all studied glasses meet the international standards for materials which can be used in the immobilization of nuclear wastes. (author)

  5. Migration-driven aggregate behaviors of human mesenchymal stem cells on a dendrimer-immobilized surface direct differentiation toward a cardiomyogenic fate commitment.

    Science.gov (United States)

    Ogawa, Yuuki; Kim, Mee-Hae; Kino-Oka, Masahiro

    2016-11-01

    Dynamic behaviors of cell aggregates on a dendrimer surface were investigated to drive the directed differentiation of human mesenchymal stem cells (hMSCs) toward a cardiomyogenic lineage. Cell aggregates on the polyamidoamine dendrimer surface with fifth-generation (G5) of dendron structure showed dynamic changes in morphology associated with repetitive stretching and contracting during migration. Spatial-temporal observations revealed cellular movement in single aggregates by their morphological change through stretching and contracting on the G5 surface, suggesting that the dynamic behavior of aggregate causes mixing of cells. However, aggregates without cell-substrate adhesions on the low-binding culture surface sustained their spherical morphology without cellular movement within a single aggregate. Furthermore, β-catenin was observed at nuclei in aggregates on the G5 surface, and expression of the cardiomyocyte marker cardiac Troponin T (cTnT) was detected. However, β-catenin localized to the nuclei only in the outer region of the aggregate on the low-binding culture surface, and cTnT expression was restricted at the exterior surface of the aggregates. These observations indicate that cell mixing within aggregates on the G5 surface induced the directed differentiation of hMSCs toward a cardiomyogenic lineage by nuclear translocation of β-catenin through dissociation of cell-cell adhesions. These results suggest that migration-driven aggregate behaviors on the dendrimer surface caused repeated morphological changes of aggregate through stretching and contracting, leading to the directed differentiation of hMSCs toward a cardiomyogenic fate commitment. Copyright © 2016 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  6. Mechanism study of dual-frequency ultrasound assisted enzymolysis on rapeseed protein by immobilized Alcalase.

    Science.gov (United States)

    Wang, Bei; Meng, Tingting; Ma, Haile; Zhang, Yanyan; Li, Yunliang; Jin, Jian; Ye, Xiaofei

    2016-09-01

    The mechanism of ultrasound field promoting enzymolysis efficiency is difficult to study, because the reaction system mixes with enzymes, proteins and hydrolysates. Immobilized enzyme is a good option that can be used to investigate the mechanism by separating enzymes out from the system after enzymolysis. The objective of this study was by using immobilized Alcalase to investigate the effects and mechanisms of the promotion of dual-frequency ultrasound (DFU) assisted-enzymolysis on rapeseed protein. Based on single factor experiments, response surface methodology model with three factors - hydrolysis time, power density and solid-liquid ratio at three levels was utilized to optimize the degree of hydrolysis (DH). Circular dichroism (CD) was used to analyze the secondary structure change of the protein, scanning electron microscopy (SEM) was used to analyze the surface microstructure change of the enzyme. The results showed that with DFU assisted-enzymolysis, the DH increased by 74.38% at the optimal levels for power density 57W/L, solid-liquid ratio 5.3g/L and enzymolysis time 76min. After DFU assisted-enzymolysis, the yield of soluble solids content, including protein, peptides and total sugar in hydrolysate increased by 64.61%, 40.88% and 23.60%, respectively. CD analysis showed that after DFU assisted-enzymolysis, the number of α-helix and random coil decreased by 10.7% and 4.5%, β-chain increased by 2.4%. SEM showed that the degree of surface roughness of immobilized Alcalase increased. The above results indicated that the improvement of hydrolysis by DFU assisted-enzymolysis was achieved by enhancing the solid solubility, changing the molecular structure of protein and increased the surface area of immobilized enzyme. Copyright © 2016 Elsevier B.V. All rights reserved.

  7. Evidence for metal-surface interactions and their role in stabilizing well-defined immobilized Ru-NHC alkene metathesis catalysts.

    Science.gov (United States)

    Samantaray, Manoja K; Alauzun, Johan; Gajan, David; Kavitake, Santosh; Mehdi, Ahmad; Veyre, Laurent; Lelli, Moreno; Lesage, Anne; Emsley, Lyndon; Copéret, Christophe; Thieuleux, Chloé

    2013-02-27

    Secondary interactions are demonstrated to direct the stability of well-defined Ru-NHC-based heterogeneous alkene metathesis catalysts. By providing key stabilization of the active sites, higher catalytic performance is achieved. Specifically, they can be described as interactions between the metal center (active site) and the surface functionality of the support, and they have been detected by surface-enhanced (1)H-(29)Si NMR spectroscopy of the ligand and (31)P solid-state NMR of the catalyst precursor. They are present only when the metal center is attached to the surface via a flexible linker (a propyl group), which allows the active site to either react with the substrate or relax, reversibly, to the surface, thus providing stability. In contrast, the use of a rigid linker (here mesitylphenyl) leads to a well-defined active site far away from the surface, stabilized only by a phosphine ligand which under reaction conditions leaves probably irreversibly, leading to faster decomposition and deactivation of the catalysts.

  8. Metabolic alkalosis during immobilization in monkeys (M. nemestrina)

    Science.gov (United States)

    Young, D. R.; Yeh, I.; Swenson, R. S.

    1983-01-01

    The systemic and renal acid-base response of monkeys during ten weeks of immobilization was studied. By three weeks of immobilization, arterial pH and bicarbonate concentrations were elevated (chronic metabolic alkalosis). Net urinary acid excretion increased in immobilized animals. Urinary bicarbonate excretion decreased during the first three weeks of immobilization, and then returned to control levels. Sustained increases in urinary ammonium excretion were seen throughout the time duration of immobilization. Neither potassium depletion nor hypokalemia was observed. Most parameters returned promptly to the normal range during the first week of recovery. Factors tentatively associated with changes in acid-base status of monkeys include contraction of extracellular fluid volume, retention of bicarbonate, increased acid excretion, and possible participation of extrarenal buffers.

  9. Development of a thiol-ene based screening platform for enzyme immobilization demonstrated using horseradish peroxidase

    DEFF Research Database (Denmark)

    Hoffmann, Christian; Pinelo, Manuel; Woodley, John

    2017-01-01

    Efficient immobilization of enzymes on support surfaces requires an exact match between the surface chemistry and the specific enzyme. A successful match would normally be identified through time consuming screening of conventional resins in multiple experiments testing individual immobilization...... strategies. In this study we present a versatile strategy that largely expands the number of possible surface functionalities for enzyme immobilization in a single, generic platform. The combination of many individual surface chemistries and thus immobilization methods in one modular system permits faster...... functionalization by thiol-ene chemistry (TEC) resulted in the formation of a functional monolayer in each well, whereas, polymer surface grafts were introduced through surface chain transfer free radical polymerization (SCT-FRP). Enzyme immobilization on the modified surfaces was evaluated by using a rhodamine...

  10. Uranium Immobilization in Wetland Soils

    Science.gov (United States)

    Jaffe, Peter R.; Koster van Groos, Paul G.; Li, Dien; Chang, Hyun-Shik; Seaman, John C.; Kaplan, Daniel I.; Peacock, Aaron D.; Scheckel, Kirk

    2014-05-01

    In wetlands, which are a major feature at the groundwater-surface water interface, plants deliver oxygen to the subsurface to keep root tissue aerobic. Some of this oxygen leaches into the rhizosphere where it will oxidize iron that typically precipitates on or near roots. Furthermore, plans provide carbon via root exudates and turnover, which in the presence of the iron oxides drives the activity of heterotrophic iron reducers in wetland soils. Oxidized iron is an important electron acceptor for many microbially-driven transformations, which can affect the fate and transport of several pollutants. It has been shown that heterotrophic iron reducing organisms, such as Geobacter sp., can reduce water soluble U(VI) to insoluble U(IV). The goal of this study was to determine if and how iron cycling in the wetland rhizosphere affects uranium dynamics. For this purpose, we operated a series of small-scale wetland mesocosms in a greenhouse to simulate the discharge of uranium-contaminated groundwater to surface waters. The mesocosms were operated with two different Fe(II) loading rates, two plant types, and unplanted controls. The mesocosms contained zones of root exclusion to differentiate between the direct presence and absence of roots in the planted mesocosms. The mesocosms were operated for several month to get fully established, after which a U(VI) solution was fed for 80 days. The mesocosms were then sacrificed and analyzed for solid-associated chemical species, microbiological characterization, micro-X-ray florescence (µ-XRF) mapping of Fe and U on the root surface, and U speciation via X-ray Absorption Near Edge Structure (XANES). Results showed that bacterial numbers including Geobacter sp., Fe(III), as well as total uranium, were highest on roots, followed by sediments near roots, and lowest in zones without much root influence. Results from the µ-XRF mapping on root surfaces indicated a strong spatial correlation between Fe and U. This correlation was

  11. Surface modification of Fe{sub 2}O{sub 3}/Fe{sub 3}O{sub 4} nanocomposites for use in immobilization of glucose oxidase; Modificacao da superficie de nanocompositos de Fe{sub 2}O{sub 3}/Fe{sub 3}O{sub 4} visando seu uso para imobilizacao da glicose oxidase

    Energy Technology Data Exchange (ETDEWEB)

    Albuquerque, I.L.T.; Santos, P.T.A.; Costa, A.C.F.M., E-mail: izabelleliz@hotmail.com, E-mail: patytaraujo@gmail.com, E-mail: ana.costa@ufcg.edu.br [Universidade Federal de Campina Grande (UFCG), PB (Brazil). Dept. de Engenharia de Materiais; Cornejo, D.R., E-mail: daniel.r.cornejo@gmail.com [Universidade de Sao Paulo (USP), SP (Brazil). Inst. de Fisica; Bicalho, S.M.C.M., E-mail: dbrandao@jhs.med.br [JHS Lab. Quimico, Sabara, MG (Brazil); Oliveira, L.S.C., E-mail: libiaconrado@yahoo.com.br [Universidade Federal de Campina Grande (UFCG), PB (Brazil). Dept. de Engenharia Quimica

    2017-04-15

    The increase in the number of people with diabetes in recent years and the high cost-benefit ratio of the existing biosensor technology have increased the interest for the development of glucose detection biosensor based on immobilization of glucose-oxidase (GOD) mainly using magnetic nanoparticles. In this context, nanocomposites of Fe{sub 2}O{sub 3}/Fe{sub 3}O{sub 4} were prepared by combustion reaction and their surface was functionalized with 3-aminopropyltriethoxysilane via silanization reaction and with chitosan via functionalization to obtain a hybrid material that was evaluated as possible GOD immobilizer. The samples were characterized by powder X-ray diffraction, Fourier transform infrared spectroscopy, thermogravimetry, scanning electron microscopy, transmission electron microscopy, magnetic properties and in vitro cytotoxicity. The results revealed that it was possible to obtain the ferrimagnetic composite, the surface modification reduced the saturation magnetization, but maintained the ferrimagnetic characteristics, and all samples were considered non-toxic. For preliminary testing of the GOD immobilization it was revealed that the nanocomposite modified with silane and chitosan showed the better result, about 2.7 mg of immobilized GOD for 100 mg of nanocomposite, which makes this material a potential alternative to manufacture GOD biosensors. (author)

  12. Rapid protein immobilization for thin film continuous flow biocatalysis.

    Science.gov (United States)

    Britton, Joshua; Raston, Colin L; Weiss, Gregory A

    2016-08-09

    A versatile enzyme immobilization strategy for thin film continuous flow processing is reported. Here, non-covalent and glutaraldehyde bioconjugation are used to immobilize enzymes on the surfaces of borosilicate reactors. This approach requires only ng of protein per reactor tube, with the stock protein solution readily recycled to sequentially coat >10 reactors. Confining reagents to thin films during immobilization reduced the amount of protein, piranha-cleaning solution, and other reagents by ∼96%. Through this technique, there was no loss of catalytic activity over 10 h processing. The results reported here combines the benefits of thin film flow processing with the mild conditions of biocatalysis.

  13. Surface Immobilization of pH-Responsive Polymer Brushes on Mesoporous Silica Nanoparticles by Enzyme Mimetic Catalytic ATRP for Controlled Cargo Release

    Directory of Open Access Journals (Sweden)

    Hang Zhou

    2016-08-01

    Full Text Available Peroxidase mimetic catalytic atom transfer radical polymerization (ATRP was first used to install tertiary amine-functionalized polymer brushes on the surface of mesoporous silica nanoparticles (MSNs in a facile and highly efficient manner. Poly(N,N-dimethylaminoethyl methacrylate (PDMAEMA brushes-grafted MSNs were fabricated by biocompatible deuterohemin-β-Ala-His-Thr-Val-Glu-Lys (DhHP-6-catalyzed surface-initiated ATRP (SI-ATRP. The resulting organic–inorganic hybrid nanocarriers were fully characterized by Fourier transform-infrared spectroscopy (FT-IR, thermogravimetric analysis (TGA, X-ray photoelectron spectroscopy (XPS, powder X-ray diffraction (XRD, SEM, TEM, Elemental analysis, Zeta-potential, and N2 adsorption–desorption isotherms, which demonstrated the successful coating of pH-responsive polymers on the MSN surface. Rhodamine 6G (Rh6G dyes were further loaded within the mesopores of this nanocarrier, and the release of Rh6G out of MSNs in a controlled fashion was achieved upon lowing the solution pH. The electrostatic repulsion of positively-charged tertiary ammonium of PDMAEMAs in acidic environments induced the stretching out of polymer brushes on MSN surfaces, thus opening the gates to allow cargo diffusion out of the mesopores of MSNs.

  14. Fabrication of calix[4]arene derivative monolayers to control orientation of antibody immobilization.

    Science.gov (United States)

    Chen, Hongxia; Liu, Feng; Qi, Fangjie; Koh, Kwangnak; Wang, Keming

    2014-03-31

    Three calix[4]arene (Cal-4) derivatives which separately contain ethylester (1), carboxylic acid (2), and crownether (3) at the lower rim with a common reactive thiol at the upper rim were synthesized and constructed to self-assembled monolayers (SAMs) on Au films. After spectroscopic characterization of the monolayers, surface coverage and orientation of antibody immobilized on the Cal-4 derivative SAMs were studied by surface plasmon resonance (SPR) technique. Experimental results revealed that the antibody could be immobilized on the Cal-4 derivatives spontaneously. The orientation of absorbed antibody on the Cal-4 derivative SAMs is related to the SAM's dipole moment. The possible orientations of the antibody immobilized on the Cal-4 derivative 1 SAM are lying-on or side-on, while on the Cal-4 derivative 2 and Cal-4 derivative 3 head-on and end-on respectively. These experimental results demonstrate the surface dipole moment of Cal-4 derivative appears to be an important factor to antibody orientation. Cal-4 derivatives are useful in developing site direct protein chips.

  15. Fabrication of Calix[4]arene Derivative Monolayers to Control Orientation of Antibody Immobilization

    Directory of Open Access Journals (Sweden)

    Hongxia Chen

    2014-03-01

    Full Text Available Three calix[4]arene (Cal-4 derivatives which separately contain ethylester (1, carboxylic acid (2, and crownether (3 at the lower rim with a common reactive thiol at the upper rim were synthesized and constructed to self-assembled monolayers (SAMs on Au films. After spectroscopic characterization of the monolayers, surface coverage and orientation of antibody immobilized on the Cal-4 derivative SAMs were studied by surface plasmon resonance (SPR technique. Experimental results revealed that the antibody could be immobilized on the Cal-4 derivatives spontaneously. The orientation of absorbed antibody on the Cal-4 derivative SAMs is related to the SAM’s dipole moment. The possible orientations of the antibody immobilized on the Cal-4 derivative 1 SAM are lying-on or side-on, while on the Cal-4 derivative 2 and Cal-4 derivative 3 head-on and end-on respectively. These experimental results demonstrate the surface dipole moment of Cal-4 derivative appears to be an important factor to antibody orientation. Cal-4 derivatives are useful in developing site direct protein chips.

  16. Dense Layer of Bacteriophages Ordered in Alternating Electric Field and Immobilized by Surface Chemical Modification as Sensing Element for Bacteria Detection.

    Science.gov (United States)

    Richter, Łukasz; Bielec, Krzysztof; Leśniewski, Adam; Łoś, Marcin; Paczesny, Jan; Hołyst, Robert

    2017-06-14

    Faster and more sensitive environmental monitoring should be developed to face the worldwide problem of bacterial infections. To remedy this issue, we demonstrate a bacteria-sensing element that utilizes dense and ordered layers of bacteriophages specific to the given bacteria strain. We combine (1) the chemical modification of a surface to increase the surface coverage of bacteriophages (2) with an alternating electric field to greatly increase the number of properly oriented bacteriophages at the surface. Usually, in sensing elements, a random orientation of bacteriophages results in steric hindrance, which results in no more than a few percent of all receptors being available. An increased number of properly ordered phages results in the optimal performance of phage receptors, manifesting in up to a 64-fold increase in sensitivity and a limit of detection as low as 100 CFU mL -1 . Our sensing elements can be applied for selective, sensitive, and fast (15 min) bacterial detection. A well-studied pair T4 bacteriophage-bacteria Escherichia coli, was used as a model; however, the method could be adapted to prepare bacteriophage-based sensors for detection of a variety of bacterial strains.

  17. Immobilization of nanoparticles by occlusion into microbial calcite

    DEFF Research Database (Denmark)

    Skuce, Rebecca L.; Tobler, Dominique Jeanette; MacLaren, Ian

    2017-01-01

    Binding of nanoparticles (NPs) to mineral surfaces influences their transport through the environment. The potential, however, for growing minerals to immobilize NPs via occlusion (the process of trapping particles inside the growing mineral) has yet to be explored in environmentally relevant...... not influence calcite precipitation at the concentrations used here. Overall, these findings demonstrate that microbially driven mineral precipitation has potential to immobilize nanoparticles in the environment via occlusion....

  18. Pesticides in surface waters: distribution, trends, and governing factors

    Science.gov (United States)

    Larson, Steven J.; Capel, Paul D.; Majewski, Michael

    1997-01-01

    Pesticde use in agriculture and non-agriculture settings has increased dramatically over the last several decades. Concern about adverse effects on the environment and human health has spurred an enormous amount of research into their environmental behavior and fate. Pesticides in Surface Waters presents a comprehensive summary of this research. This book evaluates published studies that focus on measuring pesticide concentration. The studies chosen include peer reviewed scientific literature, government reports, laboratory studies, and those using microcosms and artificial streams and ponds. The authors used this information to develop their overview of pesticide contamination of surface waters. The exhaustive compilation of data along with the fundamental science make this book essential for those involved in pesticide use, environmental protection, water quality, and human or ecological risk assessment. Pesticides in Surface Waters covers the results of actual studies, sources of pesticides to surface water, fate and transport, and environmental significance. Hundreds of data-packed tables, maps, charts, and drawings illustrate the key points, making research and application easy and cost effective.

  19. Immobilized laccase-based biosensor for the detection of disubstituted methyl and methoxy phenols - application of Box-Behnken design with response surface methodology for modeling and optimization of performance parameters.

    Science.gov (United States)

    Sarika, C; Rekha, K; Narasimha Murthy, B

    2016-11-01

    An amperometric principle-based biosensor, employing immobilized laccase enzyme from Trametes versicolor, was developed for the detection of disubstituted methyl and methoxy phenols. Three immobilization methods such as entrapment, cross-linking, and co-cross-linking, with bovine serum albumin (BSA) on nylon membrane have been compared. Among tested methods of immobilization, co-cross-linking method with BSA was superior to the other methods in terms of; sensitivity, limit of detection, response time, and operating stability. The increased sensitivity of the probe optimization of concentrations of laccase, BSA and glutaraldehyde can be achieved by, employing the Box-Behnken design of experiment.

  20. Studeis on the immobilization of water soluble phosphatic fertilizer in some soils with 32P

    International Nuclear Information System (INIS)

    Zhang Yumei; Li Rensheng; Xu Xinyu

    1985-01-01

    Using superphosphate lablled with 32 P, we studied immobilization of water-soluble phosphatic fertilizer on 12 typies of soil. The experimental result showed that major factors to govern immobilization of water-soluble phosphatic fertilizer are: quickly availible Fe that showed positive correlation with the immobilization when it was 4.64-65.72 ppm; and pH that showed negative correlation with the immobilization when it was between 5.35 and 8.88. CaCO 3 and organic matter showed a great effect on the immobilization though there wasn't obvious correlation among them

  1. Evaluation of supports and methods for immobilization of enzyme cyclodextringlycosyltransferase.

    Science.gov (United States)

    Sobral, Keli A; Rodrigues, Regina O; Oliveira, Rogério D; Olivo, José E; de Moraes, Flávio F; Zanin, Gisella M

    2003-01-01

    An experimental design with factorial planning was used for the immobilization of the enzyme cyclodextringlycosyltransferase (CGTase) from Bacillus firmus (strain no. 37) to select the best combination of support, method of immobilization, and conditions that gives primarily higher average values for the specific immobilized enzyme activity, and secondarily, higher average values for the percentage of protein fixation. The experimental design factors were as follows: supports-controlled-pore silica, chitosan, and alumina; immobilization methods-adsorption, and two covalent bonding methods, either with gamma-aminopropyltriethoxysilane or hexamethylenediamine (HEMDA); conditions-7 degrees C without agitation and 26 degrees C with stirring. The best combination of factors that lead to higher average values of the response variables was obtained with immobilization of CGTase in silica with HEMDA at 7 degrees C. However, immobilization in chitosan at 7 degrees C gave the highest immobilized CGTase specific activity, 0.25 micromole of beta-CD/ (min mg protein). Physical adsorption gave low specific enzyme activities, and, in general, a high load of enzyme leads to lower specific enzyme activity.

  2. Comparison of covalent and noncovalent immobilization of Malatya apricot pectinesterase (Prunus armeniaca L.).

    Science.gov (United States)

    Karakuş, Emine; Pekyardımcı, Sule

    2012-02-01

    Pectinesterase isolated from Malatya apricot pulp was noncovalently and covalently immobilized onto bentonite and glutaraldehyde-containing amino group functionalized porous glass beads surface at pH 8.0 and pH 9.0, respectively. The effect of various parameters such as pH, temperature, activation energy, heat and storage stability on immobilized enzyme were investigated. The optimum temperature of covalently and noncovalently immobilized PE was 50°C. This value was 60°C for free PE. Although optimum pH of covalently-immobilized PE was 8.0, this parameter was 9.0 for free and covalently-immobilized PE. The noncovalently immobilized enzyme exhibited better thermostability than the free and covalently immobilized PE.

  3. Effects of immobilization on spermiogenesis

    Science.gov (United States)

    Meitner, E. R.

    1980-01-01

    The influence of immobilization stress on spermiogenesis in rats was investigated. After 96 hour immobilization, histological changes began to manifest themselves in the form of practically complete disappearance of cell population of the wall of seminiferous tubule as well as a markedly increased number of cells with pathologic mitoses. Enzymological investigations showed various changes of activity (of acid and alkaline phosphatase and nonspecific esterase) in the 24, 48, and 96 hour immobilization groups.

  4. Immobilization thresholds of electrofishing relative to fish size

    Science.gov (United States)

    Dolan, C.R.; Miranda, L.E.

    2003-01-01

    Fish size and electrical waveforms have frequently been associated with variation in electrofishing effectiveness. Under controlled laboratory conditions, we measured the electrical power required by five electrical waveforms to immobilize eight fish species of diverse sizes and shapes. Fish size was indexed by total body length, surface area, volume, and weight; shape was indexed by the ratio of body length to body depth. Our objectives were to identify immobilization thresholds, elucidate the descriptors of fish size that were best associated with those immobilization thresholds, and determine whether the vulnerability of a species relative to other species remained constant across electrical treatments. The results confirmed that fish size is a key variable controlling the immobilization threshold and further suggested that the size descriptor best related to immobilization is fish volume. The peak power needed to immobilize fish decreased rapidly with increasing fish volume in small fish but decreased slowly for fish larger than 75-100 cm 3. Furthermore, when we controlled for size and shape, different waveforms did not favor particular species, possibly because of the overwhelming effect of body size. Many of the immobilization inconsistencies previously attributed to species might simply represent the effect of disparities in body size.

  5. Immobilization of maltase from Saccharomyces cerevisiae on thiosulfonate supports

    Directory of Open Access Journals (Sweden)

    Mihailović Mladen

    2016-01-01

    Full Text Available In this study, two commercial supports (Eupergit® C and Purolite® A109 were chemically modified in order to introduce thiosulfonate groups, which could subsequently exclusively react with cysteine residues on enzyme surface. Thereafter, the immobilization of maltase from Saccharomyces cerevisiae onto obtained thiosulfonate-activated supports was performed, resulting in high expressed enzymatic activities (around 50%, while on the other hand, immobilization on unmodified supports yielded expressed activities less than 5%. Moreover, protein loadings up to 12.3 mg g-1 and immobilized activities up to 3580 IU g-1 were achieved by employment of theses thiosulfonate supports. Desorption experiments, performed on samples taken during immobilization, proved that immobilization on thiosulfonate supports encompass first step of fast adsorption on support and second slower step of the covalent bond formation between thiosulfonate groups and thiol groups of cysteine. More importantly, although enzyme coupling occurs via covalent bond formation, performed immobilization proved to be reversible, since it was shown that 95% of immobilized activity can be detached from support after treatment with thiol reagent (β-mercaptoethanol, thus support can be reused after enzyme inactivation. [Projekat Ministarstva nauke Republike Srbije, br. III 46010

  6. Modeling wind adjustment factor and midflame wind speed for Rothermel's surface fire spread model

    Science.gov (United States)

    Patricia L. Andrews

    2012-01-01

    Rothermel's surface fire spread model was developed to use a value for the wind speed that affects surface fire, called midflame wind speed. Models have been developed to adjust 20-ft wind speed to midflame wind speed for sheltered and unsheltered surface fuel. In this report, Wind Adjustment Factor (WAF) model equations are given, and the BehavePlus fire modeling...

  7. Photocatalytic ozonation of urban wastewater and surface water using immobilized TiO2 with LEDs: Micropollutants, antibiotic resistance genes and estrogenic activity.

    Science.gov (United States)

    Moreira, Nuno F F; Sousa, José M; Macedo, Gonçalo; Ribeiro, Ana R; Barreiros, Luisa; Pedrosa, Marta; Faria, Joaquim L; Pereira, M Fernando R; Castro-Silva, Sérgio; Segundo, Marcela A; Manaia, Célia M; Nunes, Olga C; Silva, Adrián M T

    2016-05-01

    Photocatalytic ozonation was employed for the first time in continuous mode with TiO2-coated glass Raschig rings and light emitting diodes (LEDs) to treat urban wastewater as well as surface water collected from the supply area of a drinking water treatment plant (DWTP). Different levels of contamination and types of contaminants were considered in this work, including chemical priority substances (PSs) and contaminants of emerging concern (CECs), as well as potential human opportunistic antibiotic resistant bacteria and their genes (ARB&ARG). Photocatalytic ozonation was more effective than single ozonation (or even than TiO2 catalytic ozonation) in the degradation of typical reaction by-products (such as oxalic acid), and more effective than photocatalysis to remove the parent micropollutants determined in urban wastewater. In fact, only fluoxetine, clarithromycin, erythromycin and 17-alpha-ethinylestradiol (EE2) were detected after photocatalytic ozonation, by using solid-phase extraction (SPE) pre-concentration and LC-MS/MS analysis. In surface water, this treatment allowed the removal of all determined micropollutants to levels below the limit of detection (0.01-0.20 ng L(-1)). The efficiency of this process was then assessed based on the capacity to remove different groups of cultivable microorganisms and housekeeping (16S rRNA) and antibiotic resistance or related genes (intI1, blaTEM, qnrS, sul1). Photocatalytic ozonation was observed to efficiently remove microorganisms and ARGs. Although after storage total heterotrophic and ARB (to ciprofloxacin, gentamicin, meropenem), fungi, and the genes 16S rRNA and intI1, increased to values close to the pre-treatment levels, the ARGs (blaTEM, qnrS and sul1) were reduced to levels below/close to the quantification limit even after 3-days storage of treated surface water or wastewater. Yeast estrogen screen (YES), thiazolyl blue tetrazolium reduction (MTT) and lactate dehydrogenase (LDH) assays were also performed

  8. Quality factor due to roughness scattering of shear horizontal surface acoustic waves in nanoresonators

    NARCIS (Netherlands)

    Palasantzas, G.

    2008-01-01

    In this work we study the quality factor associated with dissipation due to scattering of shear horizontal surface acoustic waves by random self-affine roughness. It is shown that the quality factor is strongly influenced by both the surface roughness exponent H and the roughness amplitude w to

  9. Immobilization by Surface Conjugation of Cyclic Peptides for Effective Mimicry of the HCV-Envelope E2 Protein as a Strategy toward Synthetic Vaccines.

    Science.gov (United States)

    Meuleman, Theodorus J; Dunlop, James I; Owsianka, Anna M; van de Langemheen, Helmus; Patel, Arvind H; Liskamp, Rob M J

    2018-02-19

    Mimicry of the binding interface of antibody-antigen interactions using peptide-based modulators (i.e., epitope mimics) has promising applications for vaccine design. These epitope mimics can be synthesized in a streamlined and straightforward fashion, thereby allowing for high-throughput analysis. The design of epitope mimics is highly influenced by their spatial configuration and structural conformation. It is widely assumed that for proper mimicry sufficient conformational constraints have to be implemented. This paper describes the synthesis of bromide derivatives functionalized with a flexible TEG linker equipped with a thiol-moiety that could be used to support cyclic or linear peptides. The cyclic and linear epitope mimics were covalently conjugated via the free thiol-moiety on maleimide-activated plate surfaces. The resulting covalent, uniform, and oriented coated surface of cyclic or linear epitope mimics were subjected to an ELISA to investigate the effect of peptide cyclization with respect to mimicry of an antigen-antibody interaction of the HCV E2 glycoprotein. To the best of our knowledge, the benefit of cyclized peptides over linear peptides has been clearly demonstrated here for the first time. Cyclic epitope mimics, and not the linear epitope mimics, demonstrated specificity toward their monoclonal antibodies HC84.1 and V3.2, respectively. The described strategy for the construction of epitope mimics shows potential for high-throughput screening of key binding residues by simply changing the amino acid sequences within synthetic peptides. In this way, leucine-438 has been identified as a key binding residue for binding monoclonal antibody V3.2.

  10. Protection zones as a factor of the sustainability of immobile cultural properties in Kosovo and Metohija on the example of monasteries

    Directory of Open Access Journals (Sweden)

    Marković Gordana

    2009-01-01

    Full Text Available The protections zones around immobile cultural properties in Serbia have been determined in compliance with the Law on Cultural Properties and for the purpose of establishing a status of a cultural monument. When reviewing the established status, the coverage of a protection zone was frequently corrected, which confirms that the areas of protected natural and cultural heritage are living organisms that need a change of borders in light of a contemporary urban and regional development. The protection zones around cultural monuments have a particular significance in the context of specific conditions for the survival of the Serbian cultural and spiritual heritage in Kosovo and Metohija. A cultural property is inseparable from the space in which it is located and its full life depends on the fullness of its function, which is also an imperative of a sustainable development. At the same time, integrative protection entails a thorough and multi-disciplinary approach in the process of studying and decision-making. The establishment of protection zones is also conditioned by the inviolable attitude of the owner and user - in this case the Serbian Orthodox Church, but also by the already established status of a cultural property by the Serbian Protection Service. It is also necessary to achieve the basic security conditions in the region with a high conflict risk. In addition, there is a significant expert participation of international institutions with regards to the defining of the criteria for the establishment of the zones, and there are also tendencies of the local community in Kosovo and Metohija. The practice has confirmed difficulties in harmonizing all positions related to the establishment of zones and particularly in defining the mechanisms for the management of the zones. In those terms, it is necessary to have an integration of legal regulations and cultural policy, establishment of the widest possible cooperation at all levels and the

  11. Risk factors for hepatitis B surface antigenaemia among secondary ...

    African Journals Online (AJOL)

    2016-11-08

    Nov 8, 2016 ... adolescents and young adults between 10-24 years.7 This ... on some common risky behavioural practices that char- ... occupation, mother's level of education and risk factors for HBV infection like blood transfusion, family history of hepatitis, injections, scarification marks and tattoos, and sexual activity.

  12. Surface modification of strontium-doped porous bioactive ceramic scaffolds via poly(DOPA) coating and immobilizing silk fibroin for excellent angiogenic and osteogenic properties.

    Science.gov (United States)

    Wang, Xu; Gu, Zhipeng; Jiang, Bo; Li, Li; Yu, Xixun

    2016-04-01

    For bioceramic scaffolds employed in clinical applications, excellent bioactivity and tenacity were of great importance. Modifying inorganic SCPP scaffolds with biological macromolecules could obviously improve its bioactivity and eliminate its palpable brittleness. However, it was hard to execute directly due to extremely bad interfacial compatibility between them. In this research, dopamine (DOPA) was introduced onto strontium-doped calcium polyphosphate (SCPP) scaffolds, subsequently the preliminary material was successfully further modified by silk fibroin (SF). SCPP/D/SF possessed suitable biomechanical properties, ability to stimulate angiogenic factor secretion and excellent biocompatibility. Biomechanical examination demonstrated that SCPP/D/SF scaffolds yielded better compressive strength because of improved interfacial compatibility. MTT assay and CLSM observation showed that SCPP/D/SF scaffolds had good cytocompatibility and presented better inducing-cell-migration potential than pure SCPP scaffolds. Meanwhile, its ability to stimulate angiogenic factor secretion was measured through the ELISA assay and immunohistological analysis in vitro and in vivo respectively. The results revealed, superior to SCPP, SCPP/D/SF could effectively promote VEGF and bFGF expression, possibly leading to enhancing angiogenesis and osteogenesis. In a word, SCPP/D/SF could serve as a potential bone tissue engineering scaffold for comparable biomechanical properties and excellent bioactivity. It provided a novel idea for modification of inorganic materials to prepare promising bone tissue engineering scaffolds with the ability to accelerate bone regeneration and vascularization.

  13. Encapsulation of Platelet in Kefiran Polymer and Detection of Bioavailability of Immobilized Platelet in Probiotic Kefiran as A New Drug for Surface Bleeding

    Directory of Open Access Journals (Sweden)

    Anahita Jenab

    2015-11-01

    Full Text Available Background : Kefir contains lactic acid bacteria (Lactobacillus, Lactococcus, Leuconostoc, Acetobacter and Streptococcus and yeasts (Kluyveromyces, Torula, Candida, Saccharomyces .Kefiran is the polysaccharide produced by lactic acid bacteria in kefir.Methods : Kefiran was prepared from milk containing 0.5% fat and 10 grams kefir grains and was separated from kefir by ethanol (0.02 gram following entrapping the platelets to this polymer. Ligand of the platelet-polysaccharide was studied by FTIR.Results : FTIR results showed that the bands of C-O and C-O-C connections were formed and the polysaccharides had been attached to the receptors of the platelet glycoproteins (GP Ib,GPIIb / IIIa. Stability and encapsulation of the platelet and kefiran were assessed by Coulter Counter. Encapsulation of the platelets by polysaccharide at the beginning caused to reduce the number of platelets following by releasing of 50% of the platelets after 3 hours.Conclusion : The platelets were encapsulated in kefiran polymer and detected for bioavailability as new drug for surface bleeding. Also, kefiran has antimicrobial and antifungal properties. On the other hand, the existence of nisin in kefiran could be useful as an antibacterial lantibiotic. 

  14. Characterization of modified zeolite as microbial immobilization media on POME anaerobic digestion

    Science.gov (United States)

    Cahyono, Rochim B.; Ismiyati, Sri; Ginting, Simparmin Br; Mellyanawaty, Melly; Budhijanto, Wiratni

    2018-03-01

    As the world’s biggest palm oil producer, Indonesia generates also huge amount of Palm Oil Mill Effluent (POME) wastewater and causes serious problem in environment. In conventional method, POME was converted into biogas using lagoon system which required extensive land area. Anaerobic Fluidized Bed Reactor (AFBR) proposes more effective biogas producing with smaller land area. In the proposed system, a immobilization media would be main factor for enhancing productivity. This research studied on characterization of Lampung natural zeolite as immobilization media in the AFBR system for POME treatment. Various activation method such as physical and chemical were attempted to create more suitable material which has larger surface area, pore size distribution as well as excellent surface structures. The physical method was applied by heating up the material till 400°C while HCl was used on the chemical activation. Based on the result, the chemical activation increased the surface area significantly into 71 m2/g compared to physical as well as original zeolite. The strong acid material was quite effective to enforce the impurities within zeolite pore structure compared to heating up the material. According to distribution data, the Lampung zeolite owned the pore size with the range of 3 – 5 μm which was mesopore material. The pore size was appropriate for immobilization media as it was smaller than size of biogas microbial. The XRD patterns verified that chemical activation could maintain the zeolite structure as the original. Obviously, the SEM photograph showed apparent structure and pore size on the modified zeolite using chemical method. The testing of modified zeolite on the batch system was done to evaluate the characterization process. The modified zeolite using chemical process resulted fast reduction of COD and stabilized the volatile fatty acid as the intermediate product of anaerobic digestion, especially in the beginning of the process. Therefore, the

  15. Rapid Surface Oxidation as a Source of Surface Degradation Factor for Bi 2 Se 3

    KAUST Repository

    Kong, Desheng

    2011-06-28

    Bismuth selenide (Bi2Se3) is a topological insulator with metallic surface states (SS) residing in a large bulk bandgap. In experiments, synthesized Bi2Se3 is often heavily n-type doped due to selenium vacancies. Furthermore, it is discovered from experiments on bulk single crystals that Bi2Se3 gets additional n-type doping after exposure to the atmosphere, thereby reducing the relative contribution of SS in total conductivity. In this article, transport measurements on Bi2Se3 nanoribbons provide additional evidence of such environmental doping process. Systematic surface composition analyses by X-ray photoelectron spectroscopy reveal fast formation and continuous growth of native oxide on Bi2Se3 under ambient conditions. In addition to n-type doping at the surface, such surface oxidation is likely the material origin of the degradation of topological SS. Appropriate surface passivation or encapsulation may be required to probe topological SS of Bi2Se3 by transport measurements. © 2011 American Chemical Society.

  16. Cellulase immobilization on magnetic nanoparticles encapsulated in polymer nanospheres.

    Science.gov (United States)

    Lima, Janaina S; Araújo, Pedro H H; Sayer, Claudia; Souza, Antonio A U; Viegas, Alexandre C; de Oliveira, Débora

    2017-04-01

    Immobilization of cellulases on magnetic nanoparticles, especially magnetite nanoparticles, has been the main approach studied to make this enzyme, economically and industrially, more attractive. However, magnetite nanoparticles tend to agglomerate, are very reactive and easily oxidized in air, which has strong impact on their useful life. Thus, it is very important to provide proper surface coating to avoid the mentioned problems. This study aimed to investigate the immobilization of cellulase on magnetic nanoparticles encapsulated in polymeric nanospheres. The support was characterized in terms of morphology, average diameter, magnetic behavior and thermal decomposition analyses. The polymer nanospheres containing encapsulated magnetic nanoparticles showed superparamagnetic behavior and intensity average diameter about 150 nm. Immobilized cellulase exhibited broader temperature stability than in the free form and great reusability capacity, 69% of the initial enzyme activity was maintained after eight cycles of use. The magnetic support showed potential for cellulase immobilization and allowed fast and easy biocatalyst recovery through a single magnet.

  17. Treatment and immobilization of intermediate-level radioactive wastes

    International Nuclear Information System (INIS)

    Lerch, R.E.; Greenhalgh, W.O.; Partridge, J.A.; Richardson, G.L.

    1979-01-01

    A new program underway at the Hanford Engineering Development Laboratory (HEDL) to develop and demonstrate treatment and immobilization technologies for intermediate-level wastes (ILW) generated in the nuclear fuel cycle is discussed. ILW are defined as those liquid and solid radioactive wastes, other than high-level wastes and fuel cladding hulls, that in packaged form have radiation dose readings greater than 200 millirem/hr at the packaged surface and 10 millirem/hr at three feet from the surface. The IAEA value of 10 4 Ci/m 3 for ILW defines the upper limit. For comparative purposes, reference is also made to certain aspects of low-level radioactive wastes (LLW). Initial work has defined the sources, quantities and types of wastes which comprise ILW. Because of the wide differences in composition (e.g., acids, salt solutions, resins and zeolites, HEPA filters, etc.) the wastes may require different treatments, particularly those wastes containing volatile contaminants. The various types of ILW have been grouped into categories amenable to similar treatment. Laboratory studies are underway to define treatment technologies for liquid ILW which contain volatile contaminants and to define immobilization parameters for the residues resulting from treatment of ILW. Immobilization agents initially being evaluated for the various residues include cement, urea-formaldehyde, and bitumen although other immobilization agents will be studied. The program also includes development of acceptable test procedures for the final immobilized products as well as development of proposed criteria for storage, transportation, and disposal of the immobilized ILW

  18. Pigment Production from Immobilized Monascus sp. Utilizing Polymeric Resin Adsorption

    OpenAIRE

    Evans, Patrick J.; Wang, Henry Y.

    1984-01-01

    Pigment production by the fungus Monascus sp. was studied to determine why Monascus sp. provides more pigment in solid culture than in submerged culture. Adding a sterilized nonionic polymeric adsorbent resin directly to the growing submerged culture did not enhance the pigment production, thus indicating that pigment extraction is probably not a factor. Monascus cells immobilized in hydrogel were studied and exhibited decreased pigment production as a result of immobilization. This result is...

  19. Iodine immobilization in apatites

    International Nuclear Information System (INIS)

    Audubert, F.; Lartigue, J.E.

    2000-01-01

    In the context of a scientific program on long-lived radionuclide conditioning, a matrix for iodine 129 immobilization has been studied. A lead vanado-phosphate apatite was prepared from the melt of lead vanado-phosphate Pb 3 (VO 4 ) 1.6 (PO 4 ) 0.4 and lead iodide PbI 2 in stoichiometric proportions by calcination at 700 deg. C during 3 hours. Natural sintering of this apatite is not possible because the product decomposition occurs at 400 deg. C. Reactive sintering is the solution. The principle depends on the coating of lead iodide with lead vanado-phosphate. Lead vanado-phosphate coating is used as iodo-apatite reactant and as dense covering to confine iodine during synthesis. So the best condition to immobilize iodine during iodo-apatite synthesis is a reactive sintering at 700 deg. C under 25 MPa. We obtained an iodo-apatite surrounded with dense lead vanadate. Leaching behaviour of the matrix synthesized by solid-solid reaction is under progress in order to determine chemical durability, basic mechanisms of the iodo-apatite alteration and kinetic rate law. Iodo-apatite dissolution rates were pH and temperature dependent. We obtained a rate of 2.5 10 -3 g.m -2 .d -1 at 90 deg. C in initially de-ionised water. (authors)

  20. Covalent immobilization of lipases on monodisperse magnetic microspheres modified with PAMAM-dendrimer

    Energy Technology Data Exchange (ETDEWEB)

    Zhu, Weiwei [Lanzhou University, State Key Laboratory of Applied Organic Chemistry, Key Laboratory of Nonferrous Metal Chemistry and Resources Utilization of Gansu Province, College of Chemistry and Chemical Engineering, Institute of Biochemical Engineering and Environmental Technology (China); Zhang, Yimei [Suzhou Research Academy of North China Electric Power University (China); Hou, Chen; Pan, Duo; He, Jianjun; Zhu, Hao, E-mail: zhuhao07@lzu.edu.cn [Lanzhou University, State Key Laboratory of Applied Organic Chemistry, Key Laboratory of Nonferrous Metal Chemistry and Resources Utilization of Gansu Province, College of Chemistry and Chemical Engineering, Institute of Biochemical Engineering and Environmental Technology (China)

    2016-02-15

    This paper reported an immobilization of Candida rugosa lipase (CRL) onto PAMAM-dendrimer-grafted magnetic nanoparticles synthesized by a modified solvothermal reduction method. The dendritic magnetic nanoparticles were amply characterized by several instrumental measurements, and the CRL was covalently anchored on the three generation supports with glutaraldehyde as coupling reagent. The amount of immobilized enzyme was up to 150 mg/g support and the factors related with the enzyme activity were investigated. The immobilization of lipase improved their performance in wider ranges of pH and temperature. The immobilized lipase exhibited excellent thermal stability and reusability in comparison with free enzyme and can be reused 10 cycles with the enzymatic activity remained above 90 %. The properties of lipase improved obviously after being immobilized on the dendritic supports. The inactive immobilized lipase could be regenerated with glutaraldehyde and Cu{sup 2+}, respectively. This synthetic strategy was facile and eco-friendly for applications in lipase immobilization.

  1. The Immobilization of a Transfer Hydrogenation Catalyst on Colloidal Particles

    NARCIS (Netherlands)

    van Ravensteijn, Bas G P|info:eu-repo/dai/nl/338806008; Schild, Dirk Jan; Kegel, Willem K.|info:eu-repo/dai/nl/113729464; Klein Gebbink, Robertus J M|info:eu-repo/dai/nl/166032646

    2017-01-01

    In this paper, we report a new synthetic procedure to immobilize a transfer hydrogenation catalyst on the surface of colloidal polystyrene particles. Using supports of colloidal dimensions allows for combining a relatively high surface area for catalyst binding, mobility of the catalyst, and facile

  2. Factors affecting surface and release properties of thin PDMS films

    DEFF Research Database (Denmark)

    Vudayagiri, Sindhu; Junker, Michael Daniel; Skov, Anne Ladegaard

    2013-01-01

    Polydimethysiloxane (PDMS) elastomers are commonly used as dielectric electroactive polymers (DEAP). DEAP films are used in making actuators, generators and sensors. In the large scale manufacture of DEAP films, release of films from the substrate (carrier web) induces some defects and pre......-strain in the films which affect the overall performance of the films. The current research is directed towards investigating factors affecting the peel force and release of thin, corrugated polydimethylsiloxane films used in DEAP films. It has been shown that doping the PDMS films with small quantities...

  3. Assessing attitudes toward spinal immobilization.

    Science.gov (United States)

    Bouland, Andrew J; Jenkins, J Lee; Levy, Matthew J

    2013-10-01

    Prospective studies have improved knowledge of prehospital spinal immobilization. The opinion of Emergency Medical Services (EMS) providers regarding spinal immobilization is unknown, as is their knowledge of recent research advances. To examine the attitudes, knowledge, and comfort of prehospital and Emergency Department (ED) EMS providers regarding spinal immobilization performed under a non-selective protocol. An online survey was conducted from May to July of 2011. Participants were drawn from the Howard County Department of Fire and Rescue Services and the Howard County General Hospital ED. The survey included multiple choice questions and responses on a modified Likert scale. Correlation analysis and descriptive data were used to analyze results. Comfort using the Kendrick Extrication Device was low among ED providers. Experienced providers were more likely to indicate comfort using this device. Respondents often believed that spinal immobilization is appropriate in the management of penetrating trauma to the chest and abdomen. Reported use of padding decreased along with the frequency with which providers practice and encounter immobilized patients. Respondents often indicated that they perform spinal immobilization due solely to mechanism of injury. Providers who feel as if spinal immobilization is often performed unnecessarily were more likely to agree that immobilization causes an unnecessary delay in patient care. The results demonstrate the need for improved EMS education in the use of the Kendrick Extrication Device, backboard padding, and spinal immobilization in the management of penetrating trauma. The attitudes highlighted in this study are relevant to the implementation of a selective spinal immobilization protocol. Copyright © 2013 Elsevier Inc. All rights reserved.

  4. Effect of heparin and alendronate coating on titanium surfaces on inhibition of osteoclast and enhancement of osteoblast function

    International Nuclear Information System (INIS)

    Moon, Ho-Jin; Yun, Young-Pil; Han, Choong-Wan; Kim, Min Sung; Kim, Sung Eun; Bae, Min Soo; Kim, Gyu-Tae; Choi, Yong-Suk; Hwang, Eui-Hwan; Lee, Joon Woo; Lee, Jin-Moo; Lee, Chang-Hoon; Kim, Duck-Su; Kwon, Il Keun

    2011-01-01

    Highlights: → We examine bone metabolism of engineered alendronate attached to Ti surfaces. → Alendronate-immobilized Ti enhances activation of osteoblast differentiation. → Alendronate-immobilized Ti inhibits osteoclast differentiation. → Alendronate-immobilized Ti may be a bioactive implant with dual functions. -- Abstract: The failure of orthopedic and dental implants has been attributed mainly to loosening of the implant from host bone, which may be due to weak bonding of the implant material to bone tissue. Titanium (Ti) is used in the field of orthopedic and dental implants because of its excellent biocompatibility and outstanding mechanical properties. Therefore, in the field of materials science and tissue engineering, there has been extensive research to immobilize bioactive molecules on the surface of implant materials in order to provide the implants with improved adhesion to the host bone tissue. In this study, chemically active functional groups were introduced on the surface of Ti by a grafting reaction with heparin and then the Ti was functionalized by immobilizing alendronate onto the heparin-grafted surface. In the MC3T3-E1 cell osteogenic differentiation study, the alendronate-immobilized Ti substrates significantly enhanced alkaline phosphatase activity (ALP) and calcium content. Additionally, nuclear factor kappa B ligand (RANKL)-induced osteoclast differentiation of RAW264.7 cells was inhibited with the alendronate-immobilized Ti as confirmed by TRAP analysis. Real time PCR analysis showed that mRNA expressions of osteocalcin and osteopontin, which are markers for osteogenesis, were upregulated in MC3T3-E1 cells cultured on alendronate-immobilized Ti. The mRNA expressions of TRAP and Cathepsin K, markers for osteoclastogenesis, in RAW264.7 cells cultured on alendronate-immobilized Ti were down-regulated. Our study suggests that alendronate-immobilized Ti may be a bioactive implant with dual functions to enhance osteoblast

  5. CORROSION AND SURFACE PROTECTION IN MACHINE MATERIALS FRICTION HAVE DIFFERENT SURFACE PAIRS EXPERIMENTAL INVESTIGATION OF FACTORS

    Directory of Open Access Journals (Sweden)

    Senai YALCINKAYA

    2017-05-01

    Full Text Available Friction force, normal force, linear change. The normal force varies with the loads on the friction object. In order to determine the friction force and the friction coefficient, the friction object and the friction speed are used. The experimental work was carried out in three stages. In the first stage, the effect of normal force on the friction force was studied. In the second step, the friction force of the friction surface area is influenced. The effect of the change of the shear rate in step 3 on the friction force was investigated. At the last stage, the experimental study of the effect of the material selection on the friction force was made and it was seen that the aluminum / brass surface pair had the smallest friction coefficient as a result of the opening. The greatest coefficient of friction is found in the pair of glass / felt objects.

  6. CORROSION AND SURFACE PROTECTION IN MACHINE MATERIALS FRICTION HAVE DIFFERENT SURFACE PAIRS EXPERIMENTAL INVESTIGATION OF FACTORS

    OpenAIRE

    Senai YALCINKAYA

    2017-01-01

    Friction force, normal force, linear change. The normal force varies with the loads on the friction object. In order to determine the friction force and the friction coefficient, the friction object and the friction speed are used. The experimental work was carried out in three stages. In the first stage, the effect of normal force on the friction force was studied. In the second step, the friction force of the friction surface area is influenced. The effect of the change of the s...

  7. Shielding Factors for Gamma Radiation from Activity Deposited on Structures and Ground Surfaces

    DEFF Research Database (Denmark)

    Jensen, Per Hedemann

    1985-01-01

    -source strength. The radiation sources considered in the model are fallout radioactivity deposited on roofs, outer walls, and ground surfaces. For any combination of source strength on roof, outer wall, and ground surface, the model calculates shielding factors for specified photon energies. The input data...... are the dimensions of the house, the thickness of the walls and floors, the window dimensions, and the size of the surrounding ground surface. The fallout source strength on the surfaces is allowed to have different values due to different deposition velocities to these surfaces. This feature of the model also makes...

  8. Remote handling in the Plutonium Immobilization Project -- Second stage immobilization

    International Nuclear Information System (INIS)

    Kriikku, E.

    1999-01-01

    The Savannah River Site (SRS) will immobilize excess plutonium in ceramic pucks and seal the pucks inside welded cans. Automated equipment will place these cans in magazines and the magazines in a Defense Waste Processing Facility (DWPF) canister. The DWPF will fill the canister with glass for permanent storage. Due to the radiation, remote equipment will perform these operations in a contained environment. The Plutonium Immobilization Project is in the conceptual design stage and the facility will begin operation in 2008. This paper discusses the Plutonium Immobilization Project phase 2 automation equipment conceptual design, equipment design, and work completed

  9. Factors Affecting Optimal Surface Roughness of AISI 4140 Steel in Turning Operation Using Taguchi Experiment

    Science.gov (United States)

    Novareza, O.; Sulistiyarini, D. H.; Wiradmoko, R.

    2018-02-01

    This paper presents the result of using Taguchi method in turning process of medium carbon steel of AISI 4140. The primary concern is to find the optimal surface roughness after turning process. The taguchi method is used to get a combination of factors and factor levels in order to get the optimum surface roughness level. Four important factors with three levels were used in experiment based on Taguchi method. A number of 27 experiments were carried out during the research and analysed using analysis of variance (ANOVA) method. The result of surface finish was determined in Ra type surface roughness. The depth of cut was found to be the most important factors for reducing the surface roughness of AISI 4140 steel. On the contrary, the other important factors i.e. spindle speed and rake side angle of the tool were proven to be less factors that affecting the surface finish. It is interesting to see the effect of coolant composition that gained the second important factors to reduce the roughness. It may need further research to explain this result.

  10. Application, chemistry, and environmental implications of contaminant-immobilization amendments on agricultural soil and water quality.

    Science.gov (United States)

    Udeigwe, Theophilus K; Eze, Peter N; Teboh, Jasper M; Stietiya, Mohammed H

    2011-01-01

    Contaminants such as nitrogen (N), phosphorus (P), dissolved organic carbon (DOC), arsenic (As), heavy metals, and infectious pathogens are often associated with agricultural systems. Various soil and water remediation techniques including the use of chemical amendments have been employed to reduce the risks associated with these contaminants. This paper reviews the use of chemical amendments for immobilizing principal agricultural contaminants, the chemistry of contaminant immobilization, and the environmental consequences associated with the use of these chemical products. The commonly used chemical amendments were grouped into aluminum-, calcium-, and iron-containing products. Other products of interest include phosphorus-containing compounds and silicate clays. Mechanisms of contaminant immobilization could include one or a combination of the following: surface precipitation, adsorption to mineral surfaces (ion exchange and formation of stable complexes), precipitation as salts, and co-precipitation. The reaction pH, redox potential, clay minerals, and organic matter are potential factors that could control contaminant-immobilization processes. Reviews of potential environmental implications revealed that undesirable substances such as trace elements, fluoride, sulfate, total dissolved solids, as well as radioactive materials associated with some industrial wastes used as amendment could be leached to ground water or lost through runoff to receiving water bodies. The acidity or alkalinity associated with some of the industrial-waste amendments could also constitute a substantial environmental hazard. Chemical amendments could introduce elements capable of inducing or affecting the activities of certain lithotrophic microbes that could influence vital geochemical processes such as mineral dissolution and formation, weathering, and organic matter mineralization. Published by Elsevier Ltd.

  11. Impact of growth environment and physiological state on metal immobilization by Pseudomonas aeruginosa PAO1.

    Science.gov (United States)

    Hunter, Ryan C; Phoenix, Vernon R; Saxena, Anuradha; Beveridge, Terry J

    2010-07-01

    Environmental growth conditions and cell physiology have the potential to influence bacterial surface-metal interactions in both planktonic and biofilm systems. Here, Pseudomonas aeruginosa was studied to determine the influence of these factors (pH, redox potential, and active respiration) on surface electrostatics and metal immobilization. Acid-base titrations revealed a decrease in ionizable ligands at pKa 5 (putative carboxyls) in cells grown below pH 6.2 and in cells grown anaerobically relative to cells grown under oxic and circumneutral pH conditions. This observation correlates with Western immunoblotting assays that revealed a reduction in carboxylated B-band lipopolysaccharide in these cells. Furthermore, spectrophotometric analysis revealed a decrease in zinc, copper, and iron immobilization in these cells, suggesting that lipopolysaccharide modification in response to environmental stimuli influences metal binding. The effect of active versus inactive metabolism on metal adsorption was also examined using respiration inhibitors carbonyl cyanide m-chlorophenylhydrazone and sodium azide. Cells treated with these compounds bound more zinc, copper, and iron than untreated controls, suggesting proton extrusion through respiration competes with metal cations for reactive groups on the cell surface. Accumulation of gold did not show the same trend, and transmission electron microscopy studies confirmed it was not a surface-mediated process. These results suggest that variations in growth environment and cell physiology influence metal accumulation by bacterial cell surfaces and may help to explain discontinuous accumulation of metal observed throughout microbial communities.

  12. Dependence of Ideality Factor in Lateral PNP Transistors on Surface Carrier Concentration

    Science.gov (United States)

    Li, Xingji; Yang, Jianqun; Barnaby, Hugh J.; Galloway, Kenneth F.; Schrimpf, Ronald D.; Fleetwood, Daniel M.; Liu, Chaoming

    2017-06-01

    The influence of surface carrier concentration on the ideality factor of excess base current (ΔIB) in gated lateral PNP (GLPNP) bipolar junction transistors (BJTs) induced by 1-MeV electrons is investigated. ΔIB in LPNP BJTs is impacted by the surface carrier density and radiation-induced interface traps. In GLPNP BJTs, the surface carrier concentration can be controlled by the voltage applied to a gate over the base region. The ideality factor changes after irradiation, and its dependence on emitter-base voltage (VEB) is a function of gate voltage. For the irradiated devices, as the gate voltage decreases from +20 to -5 V, the ideality factor for excess base current changes from a single slope to two-slope behavior. The majority carrier concentration at the surface of the base, controlled by the gate voltage, impacts the excess base current and its ideality factor.

  13. Evaluation of oriented lysozyme immobilized with monoclonal antibody

    Science.gov (United States)

    Aoyagi, Satoka; Okada, Keigo; Shigyo, Ayako; Man, Naoki; Karen, Akiya

    2008-12-01

    The orientation of a lysozyme immobilized with a monoclonal antibody was evaluated based on determination of the uppermost surface structure using time-of-flight secondary ion mass spectrometry (TOF-SIMS). Specific peaks of the oriented lysozyme immobilized with monoclonal anti-lysozyme antibody were obtained in comparison with reference samples, non-oriented immobilized lysozyme and immobilized anti-lysozyme antibody. All samples were freeze-dried before TOF-SIMS measurement, and then each sample was measured using TOF-SIMS with a bismuth cluster ion source. TOF-SIMS spectra were analyzed to select peaks specific to the oriented immobilized lysozyme as well as to identify their chemical formula and ensemble of amino acids. The possible chemical formulae of the lysozyme fragments were then investigated with an element matching program and a residue matching program. The results from TOF-SIMS spectra analysis were compared to the amino acid sequence of the lysozyme and its three-dimensional structure registered in the protein data bank. Finally, the fragment-ion-generating regions of the oriented immobilized lysozyme were determined based on the suggested residues and the three-dimensional structure.

  14. Experimental analysis on removal factor of smear method in measurement of surface contamination

    International Nuclear Information System (INIS)

    Sugiura, Nobuyuki; Taira, Junichi; Takenaka, Keisuke; Yamanaka, Kazuo; Sugai, Kenji; Kosako, Toshiso

    2007-01-01

    The smear test is one of the important ways to measure surface contamination. The loose contamination under the high background radiation, which is more significant in handling non-sealed radioisotopes, can be evaluated by this method. The removal factor is defined as the ratio of the activity removed from the surface by one smear to the whole activity of the removable surface contamination. The removal factor is greatly changed by the quality and condition of surface materials. In this study, the values of removal factor at several typical surface conditions were evaluated experimentally and the practical application of those values was considered. It is required the smear should be pressed by moderate pressure when wiping the surface. The pressure from 1.0 kg to 1.5 kg per filter paper was recommended. The removal factor showed lower value in wiping by the pressure below 1.0 kg. The value of 0.5 for the removal factor could be applied to the smooth surface of linoleum, concrete coated with paint or epoxy resin, stainless steel and glass with the statistical allowance. (author)

  15. Hydrophilic PCU scaffolds prepared by grafting PEGMA and immobilizing gelatin to enhance cell adhesion and proliferation

    Energy Technology Data Exchange (ETDEWEB)

    Shi, Changcan; Yuan, Wenjie; Khan, Musammir; Li, Qian [School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072 (China); Feng, Yakai, E-mail: yakaifeng@tju.edu.cn [School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072 (China); Key Laboratory of Systems Bioengineering of Ministry of Education, Tianjin University, Tianjin 300072 (China); Tianjin University-Helmholtz-Zentrum Geesthacht, Joint Laboratory for Biomaterials and Regenerative Medicine, Tianjin 300072 (China); Collaborative Innovation Center of Chemical Science and Chemical Engineering (Tianjin) Tianjin 300072 (China); Yao, Fanglian [School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072 (China); Key Laboratory of Systems Bioengineering of Ministry of Education, Tianjin University, Tianjin 300072 (China); Tianjin University-Helmholtz-Zentrum Geesthacht, Joint Laboratory for Biomaterials and Regenerative Medicine, Tianjin 300072 (China); Zhang, Wencheng, E-mail: wenchengzhang@yahoo.com [Department of Physiology and Pathophysiology, Logistics University of Chinese People' s Armed Police Force, Tianjin 300162 (China)

    2015-05-01

    Gelatin contains many functional motifs which can modulate cell specific adhesion, so we modified polycarbonate urethane (PCU) scaffold surface by immobilization of gelatin. PCU-g-gelatin scaffolds were prepared by direct immobilizing gelatins onto the surface of aminated PCU scaffolds. To increase the immobilization amount of gelatin, poly(ethylene glycol) methacrylate (PEGMA) was grafted onto PCU scaffolds by surface initiated atom transfer radical polymerization. Then, following amination and immobilization, PCU-g-PEGMA-g-gelatin scaffolds were obtained. Both modified scaffolds were characterized by chemical and biological methods. After immobilization of gelatin, the microfiber surface became rough, but the original morphology of scaffolds was maintained successfully. PCU-g-PEGMA-g-gelatin scaffolds were more hydrophilic than PCU-g-gelatin scaffolds. Because hydrophilic PEGMA and gelatin were grafted and immobilized onto the surface, the PCU-g-PEGMA-g-gelatin scaffolds showed low platelet adhesion, perfect anti-hemolytic activity and excellent cell growth and proliferation capacity. It could be envisioned that PCU-g-PEGMA-g-gelatin scaffolds might have potential applications in tissue engineering artificial scaffolds. - Graphical abstract: PCU-g-gelatin scaffolds were prepared by direct immobilizing gelatin onto the surface of aminated PCU scaffolds (method a). To increase the immobilization amount of gelatin, PEGMAs were grafted onto the scaffold surface by SI-ATRP. PCU-g-PEGMA-g-gelatin scaffolds were prepared by method b. The gelatin modified scaffolds exhibited high hydrophilicity, low platelet adhesion, perfect anti-hemolytic activity, and excellent cell adhesion and proliferation capacity. They might have potential applications as tissue engineering scaffolds for artificial blood vessels. - Highlights: • Hydrophilic scaffolds were prepared by grafting PEGMA and immobilization of gelatins. • Grafting PEGMA enhanced the immobilization amount of gelatin

  16. N-Confused Porphyrin Immobilized on Solid Supports: Synthesis and Metal Ions Sensing Efficacy

    Directory of Open Access Journals (Sweden)

    Sara R. D. Gamelas

    2018-04-01

    Full Text Available In this work, the N-confused porphyrin 5,10,15,20-tetraphenyl-2-aza-21-carbaporphyrin (NCTPP was immobilized on neutral or cationic supports based on silica and on Merrifield resin. The new materials were characterized by appropriate techniques (UV-Vis spectroscopy, SEM, and zeta potential analysis. Piezoelectric quartz crystal gold electrodes were coated with the different hybrids and their ability to interact with heavy metals was evaluated. The preliminary results obtained showed that the new materials can be explored for metal cations detection and the modification of the material surface is a key factor in tuning the metal selectivity.

  17. Smectite clays as solid supports for immobilization of beta-glucosidase : Synthesis, characterization, and biochemical properties

    NARCIS (Netherlands)

    Serefoglou, Evangelia; Litina, Kiriaki; Gournis, Dimitrios; Kalogeris, Emmanuel; Tzialla, Aikaterini A.; Pavlidis, Ioannis V.; Stamatis, Haralambos; Maccallini, Enrico; Lubomska, Monika; Rudolf, Petra

    2008-01-01

    Nanomaterials as solid supports can improve the efficiency of immobilized enzymes by reducing diffusional limitation as well as by increasing the surface area per mass unit and therefore improving enzyme loading. In this work, beta-glucosidase from almonds was immobilized on two smectite nanoclays.

  18. An immobilization antigen gene of the fish-parasitic protozoan Ichthyophthirius multifiliis strain ARS-6

    Science.gov (United States)

    Ichthyophthirius multifiliis (Ich) is a severe fish parasite that causes ‘white spot’ disease in many freshwater fish and leads to high mortality. The antigens on the parasite surface are involved in the antibody-mediated immobilization and hence designated as immobilization antigens (i-antigens). ...

  19. Surface functionalization of nanoporous alumina with bone morphogenetic protein 2 for inducing osteogenic differentiation of mesenchymal stem cells

    International Nuclear Information System (INIS)

    Song, Yuanhui; Ju, Yang; Morita, Yasuyuki; Xu, Baiyao; Song, Guanbin

    2014-01-01

    Many studies have demonstrated the possibility to regulate cellular behavior by manipulating the specific characteristics of biomaterials including the physical features and chemical properties. To investigate the synergistic effect of chemical factors and surface topography on the growth behavior of mesenchymal stem cells (MSCs), bone morphorgenic protein 2 (BMP2) was immobilized onto porous alumina substrates with different pore sizes. The BMP2-immobilized alumina substrates were characterized with scanning electron microscopy (SEM) and X-ray photoelectron spectroscopy (XPS). Growth behavior and osteogenic differentiation of MSCs cultured on the different substrates were investigated. Cell adhesion and morphological changes were observed with SEM, and the results showed that the BMP2-immobilized alumina substrate was able to promote adhesion and spreading of MSCs. MTT assay and immunofluorescence staining of integrin β1 revealed that the BMP2-immobilized alumina substrates were favorable for cell growth. To evaluate the differentiation of MSCs, osteoblastic differentiation markers, such as alkaline phosphatase (ALP) activity and mineralization, were investigated. Compared with those of untreated alumina substrates, significantly higher ALP activities and mineralization were detected in cells cultured on BMP2-immobilized alumina substrates. The results suggested that surface functionalization of nanoporous alumina substrates with BMP2 was beneficial for cell growth and osteogenic differentiation. With the approach of immobilizing growth factors onto material substrates, it provided a new insight to exploit novel biofunctional materials for tissue engineering. - Highlights: • BMP2 was immobilized onto nanoporous alumina substrates with different pore sizes. • BMP2-immobilized substrates were able to promote adhesion and spreading of MSCs. • BMP2-immobilized substrates were favorable for cell growth of MSCs. • BMP2-immobilized substrates promoted osteogenic

  20. Immobilization of a human epidermal growth factor receptor 2 mimotope-derived synthetic peptide on Au and its potential application for detection of herceptin in human serum by quartz crystal microbalance.

    Science.gov (United States)

    Shang, Yuqin; Singh, Pankaj R; Chisti, Mohammad M; Mernaugh, Ray; Zeng, Xiangqun

    2011-12-01

    Therapeutic antibodies are antigenically similar to human antibodies and are difficult to detect in assays of human serum samples without the use of the therapeutic antibody's complementary antigen. Herein for the first time, we established a platform to detect Herceptin in solutions by using a small (synthetic peptide immobilized on the surface of a Au quartz electrode. We used the HER2 mimotope as a substitute for the HER2 receptor protein in piezoimmunosensor or quartz crystal microbalance (QCM) assays to detect Herceptin in human serum. We demonstrated that assay sensitivity was dependent upon the amino acids used to tether and link the peptide to the sensor surface and the buffers used to carry out the assays. The detection limit of the piezoimmunosensor assay was 0.038 nM with a linear operating range of 0.038-0.859 nM. Little nonspecific binding to other therapeutic antibodies (Avastin and Rituxan) was observed. Levels of Herceptin in serum samples obtained from treated patients, as ascertained using the synthetic peptide-based QCM assay, were typical for those treated with Herceptin. The findings of this study are significant in that low-cost synthetic peptides could be used in a QCM assay, in lieu of native or recombinant antigens or capture antibodies, to rapidly detect a therapeutic antibody in human serum. The results suggested that a synthetic peptide bearing a particular functional sequence could be applied for developing a new generation of affinity-based immunosensors to detect a broad range of clinical biomarkers.

  1. Characterization of immobilization methods of antiviral antibodies in serum for electrochemical biosensors

    Science.gov (United States)

    Huy, Tran Quang; Hanh, Nguyen Thi Hong; Van Chung, Pham; Anh, Dang Duc; Nga, Phan Thi; Tuan, Mai Anh

    2011-06-01

    In this paper, we describes different methods to immobilize Japanese encephalitis virus (JEV) antibodies in human serum onto the interdigitated surface of a microelectrode sensor for optimizing electrochemical detection: (1) direct covalent binding to the silanized surface, (2) binding to the silanized surface via a cross-linker of glutaraldehyde (GA), (3) binding to glutaraldehyde/silanized surface via goat anti-human IgG polyclonal antibody and (4) binding to glutaraldehyde/silanized surface via protein A (PrA). Field emission scanning electron microscopy, Fourier transform infrared spectrometry, and fluorescence microscopy are used to verify the characteristics of antibodies on the interdigitated surface after the serum antibodies immobilization. The analyzed results indicate that the use of protein A is an effective choice for immobilization and orientation of antibodies in serum for electrochemical biosensors. This study provides an advantageous immobilization method of serum containing antiviral antibodies to develop electrochemical biosensors for preliminary screening of viruses in clinical samples from outbreaks.

  2. Synchronous scattering and diffraction from gold nanotextured surfaces with structure factors

    Science.gov (United States)

    Gu, Min-Jhong; Lee, Ming-Tsang; Huang, Chien-Hsun; Wu, Chi-Chun; Chen, Yu-Bin

    2018-05-01

    Synchronous scattering and diffraction were demonstrated using reflectance from gold nanotextured surfaces at oblique (θi = 15° and 60°) incidence of wavelength λ = 405 nm. Two samples of unique auto-correlation functions were cost-effectively fabricated. Multiple structure factors of their profiles were confirmed with Fourier expansions. Bi-directional reflectance function (BRDF) from these samples provided experimental proofs. On the other hand, standard deviation of height and unique auto-correlation function of each sample were used to generate surfaces numerically. Comparing their BRDF with those of totally random rough surfaces further suggested that structure factors in profile could reduce specular reflection more than totally random roughness.

  3. Rhizopus oryzae lipase immobilized on hierarchical mesoporous silica supports for transesterification of rice bran oil.

    Science.gov (United States)

    Ramachandran, Prashanth; Narayanan, Guru Krupa; Gandhi, Sakthivel; Sethuraman, Swaminathan; Krishnan, Uma Maheswari

    2015-03-01

    The tunable textural properties of self-oriented mesoporous silica were investigated for their suitability as enzyme immobilization matrices to support transesterification of rice bran oil. Different morphologies of mesoporous silica (rod-like, rice-like, and spherical) were synthesized and characterized by scanning electron microscopy (SEM), transmission electron microscopy (TEM), and nitrogen adsorption-desorption isotherms. The surface area, pore size, and ordered arrangement of the pores were found to influence the immobilization and activity of the enzyme in the mesopores. The immobilization in rod-like silica was highest with an immobilization efficiency of 63 % and exhibited minimal activity loss after immobilization. Functionalization of the mesoporous surface with ethyl groups further enhanced the enzyme immobilization. The free enzyme lost most of its activity at 50 °C while the immobilized enzyme showed activity even up to 60 °C. Transesterified product yield of nearly 82 % was obtained for 24 h of reaction with enzyme immobilized on ethyl-functionalized SBA-15 at an oil:methanol ratio of 1:3. Fourier transform infrared spectroscopy (FT-IR) and Gas chromatography-mass spectrometry (GC-MS) were used to characterize the transesterified product obtained. The reusability of the immobilized enzyme was studied for 3 cycles.

  4. Graphene immobilized enzyme/polyethersulfone mixed matrix membrane: Enhanced antibacterial, permeable and mechanical properties

    International Nuclear Information System (INIS)

    Duan, Linlin; Wang, Yuanming; Zhang, Yatao; Liu, Jindun

    2015-01-01

    Graphical abstract: - Highlights: • Lysozyme was immobilized on the surface of graphene oxide (GO) and reduced GO (RGO). • The novel hybrid membranes based on lysozyme and graphene were fabricated firstly. • These membranes showed good antibacterial and mechanical performance. - Abstract: Enzyme immobilization has been developed to address lots of issues of free enzyme, such as instability, low activity and difficult to retain. In this study, graphene was used as an ideal carrier for lysozyme immobilization, including graphene oxide (GO) immobilized lysozyme (GO-Ly) and chemically reduced graphene oxide (CRGO) immobilized lysozyme (CRGO-Ly). Herein, lysozyme as a bio-antibacterial agent has excellent antibacterial performance and the products of its catalysis are safety and nontoxic. Then the immobilized lysozyme materials were blended into polyethersulfone (PES) casting solution to prepare PES ultrafiltration membrane via phase inversion method. GO and CRGO were characterized by Fourier transform infrared spectroscopy (FTIR), Ultraviolet–visible spectrum (UV), X-ray diffraction (XRD), and transmission electron microscopy (TEM) and the immobilized lysozyme composites were observed by fluorescent microscopy. The results revealed that GO and CRGO were successfully synthesized and lysozyme was immobilized on their surfaces. The morphology, hydrophilicity, mechanical properties, separation properties and antibacterial activity of the hybrid membranes were characterized in detail. The hydrophilicity, water flux and mechanical strength of the hybrid membranes were significantly enhanced after adding the immobilized lysozyme. In the antibacterial experiment, the hybrid membranes exhibited an effective antibacterial performance against Escherichia coli (E. coli).

  5. Graphene immobilized enzyme/polyethersulfone mixed matrix membrane: Enhanced antibacterial, permeable and mechanical properties

    Energy Technology Data Exchange (ETDEWEB)

    Duan, Linlin; Wang, Yuanming [School of Chemical Engineering and Energy, Zhengzhou University, Zhengzhou 450001 (China); Zhang, Yatao, E-mail: zhangyatao@zzu.edu.cn [School of Chemical Engineering and Energy, Zhengzhou University, Zhengzhou 450001 (China); UNESCO Centre for Membrane Science and Technology, University of New South Wales, Sydney, NSW 2052 (Australia); Liu, Jindun [School of Chemical Engineering and Energy, Zhengzhou University, Zhengzhou 450001 (China)

    2015-11-15

    Graphical abstract: - Highlights: • Lysozyme was immobilized on the surface of graphene oxide (GO) and reduced GO (RGO). • The novel hybrid membranes based on lysozyme and graphene were fabricated firstly. • These membranes showed good antibacterial and mechanical performance. - Abstract: Enzyme immobilization has been developed to address lots of issues of free enzyme, such as instability, low activity and difficult to retain. In this study, graphene was used as an ideal carrier for lysozyme immobilization, including graphene oxide (GO) immobilized lysozyme (GO-Ly) and chemically reduced graphene oxide (CRGO) immobilized lysozyme (CRGO-Ly). Herein, lysozyme as a bio-antibacterial agent has excellent antibacterial performance and the products of its catalysis are safety and nontoxic. Then the immobilized lysozyme materials were blended into polyethersulfone (PES) casting solution to prepare PES ultrafiltration membrane via phase inversion method. GO and CRGO were characterized by Fourier transform infrared spectroscopy (FTIR), Ultraviolet–visible spectrum (UV), X-ray diffraction (XRD), and transmission electron microscopy (TEM) and the immobilized lysozyme composites were observed by fluorescent microscopy. The results revealed that GO and CRGO were successfully synthesized and lysozyme was immobilized on their surfaces. The morphology, hydrophilicity, mechanical properties, separation properties and antibacterial activity of the hybrid membranes were characterized in detail. The hydrophilicity, water flux and mechanical strength of the hybrid membranes were significantly enhanced after adding the immobilized lysozyme. In the antibacterial experiment, the hybrid membranes exhibited an effective antibacterial performance against Escherichia coli (E. coli).

  6. Enzyme immobilization and biocatalysis of polysiloxanes

    Science.gov (United States)

    Poojari, Yadagiri

    H, temperature, cross-link density, organic solvents and storage time using a hemoglobin assay. A notable finding was that free pepsin had zero activity in neutral buffer solution (pH 7) after incubation for 5 hours, while pepsin immobilized in the silicone elastomers was found to retain more than 70% of its maximum normalized activity. These results demonstrate that cross-linked poly(dimethylsiloxane) (PDMS) is a promising support material for the physical entrapment of hydrolytic enzymes such as pepsin. The Novozym-435 has been widely employed as a biocatalyst for esterification and transesterification of a variety of organic compounds including synthesis of polyesters and polylactones due to its high catalytic-efficiency and high thermal stability in organic media. However, the Novozym-435 was found to have poor mechanical stability and the enzyme was found to leach out from the resin into the organic media. In the present research work, efforts were made to solve the above two problems by chemical immobilization of CALB on surface modified porous silica gel particles. The surface of the porous silica gel particles was silanized using (gamma-Aminopropyl)triethoxysilane and then the CALB was chemically crosslinked onto the surface of the silica gel particles using glutaraldehyde. Although the thermal stability of the CALB immobilized silica gel particles was found to be lower compared to that of Novozym-435. The CALB immobilized silica gel particles showed higher enzymatic activity and higher mechanical stability compared to that of Novozym-435.

  7. High-level-waste immobilization

    International Nuclear Information System (INIS)

    Crandall, J.L.

    1982-01-01

    Analysis of risks, environmental effects, process feasibility, and costs for disposal of immobilized high-level wastes in geologic repositories indicates that the disposal system safety has a low sensitivity to the choice of the waste disposal form

  8. Radiation exchange factors between specular inner surfaces of a rectangular enclosure such as transplant production unit

    International Nuclear Information System (INIS)

    Abdel-Ghany, Ahmed M.; Kozai, Toyoki

    2006-01-01

    General mathematical relations are presented for the specular exchange factors, F S , of diffuse radiation exchange between the inner surfaces of a rectangular enclosure. Three of these surfaces are specular reflectors, diffuse emitters and the fourth surface is a diffuse reflector, diffuse emitter. This enclosure can be used as a transplant production unit with artificial lighting for electric energy saving purposes. An image system and the crossed string method are used to derive these relations. The resulting expressions are conceptually simple and similar to the commonly known expressions of the exchange factors between diffuse surfaces, F. The accuracy of the presented F S relations was examined for different numbers of multiple reflections, N, on the specular surfaces and for different aspect ratios (ratio of the width, w to the height, h). The results proved that the relations are accurate and strongly satisfy the well-known relation of the radiation exchange between enclosure surfaces and satisfy the reciprocity relation. For any aspect ratio, considering N of 150 between highly reflective surfaces (ρ = 0.99) is sufficient to estimate the F S factors without any possible error. Using specular reflecting surfaces in such cases significantly reduces the electric energy consumption used for lighting

  9. Biosorption of americium-241 by immobilized Rhizopus arrihizus

    International Nuclear Information System (INIS)

    Liao Jiali; Yang Yuanyou; Luo Shunzhong; Liu Ning; Jin Jiannan; Zhang Taiming; Zhao Pengji

    2004-01-01

    Rhizopus arrihizus (R. arrihizus), a fungus, which in previous experiments had shown encouraging ability to remove 241 Am from solutions, was immobilized by calcium alginate and other reagents. The various factors affecting 241 Am biosorption by the immobilized R. arrihizus were investigated. The results showed that not only can immobilized R. arrihizus adsorb 241 Am as efficiently as free R. arrihizus, but that also can be used repeatedly or continuously. The biosorption equilibrium was achieved within 2 h, and more than 94% of 241 Am was removed from 241 Am solutions of 1.08 MBq/l by immobilized R. arrihizu in the pH range 1-7. Temperature did not affect the adsorption on immobilized R. arrihizus in the range 15-45 deg. C. After repeated adsorption for 8 times, the immobilized R. arrihizus still adsorbed more than 97% of 241 Am. At this time, the total adsorption of 241 Am was more than 88.6 KBq/g, and had not yet reached saturation. Ninety-five percent of the adsorbed 241 Am was desorbed by saturated EDTA solution and 98% by 2 mol/l HNO 3

  10. Biosorption of americium-241 by immobilized Rhizopus arrihizus

    Energy Technology Data Exchange (ETDEWEB)

    Liao Jiali E-mail: liaojiali@163.com; Yang Yuanyou; Luo Shunzhong; Liu Ning; Jin Jiannan; Zhang Taiming; Zhao Pengji

    2004-01-01

    Rhizopus arrihizus (R. arrihizus), a fungus, which in previous experiments had shown encouraging ability to remove {sup 241}Am from solutions, was immobilized by calcium alginate and other reagents. The various factors affecting {sup 241}Am biosorption by the immobilized R. arrihizus were investigated. The results showed that not only can immobilized R. arrihizus adsorb {sup 241}Am as efficiently as free R. arrihizus, but that also can be used repeatedly or continuously. The biosorption equilibrium was achieved within 2 h, and more than 94% of {sup 241}Am was removed from {sup 241}Am solutions of 1.08 MBq/l by immobilized R. arrihizu in the pH range 1-7. Temperature did not affect the adsorption on immobilized R. arrihizus in the range 15-45 deg. C. After repeated adsorption for 8 times, the immobilized R. arrihizus still adsorbed more than 97% of {sup 241}Am. At this time, the total adsorption of {sup 241}Am was more than 88.6 KBq/g, and had not yet reached saturation. Ninety-five percent of the adsorbed {sup 241}Am was desorbed by saturated EDTA solution and 98% by 2 mol/l HNO{sub 3}.

  11. lnvestigation of Patch Coatings lnfluence on the Stress lntensity Factor for Surface Cracks

    DEFF Research Database (Denmark)

    Lambertsen, Søren Heide; Jepsen, Michael S.; Damkilde, Lars

    2013-01-01

    layer on the surface. The stress intensity factor is computed by means of the J-integral and the FE-model is setup with a nonlinear material model to establish the upper boundary for the patch stress capacity. A reduction of the stress intensity factor of approximately 2 % and a reduction of the crack...... to damp the free end of the crack, which will reduce the stress intensity factor and subsequently reduce the crack growth. The considered patch consists of a surface layer of 0.2 mm two component adhesive Epoxy, 3M DP 460. The models described in this work contains five different crack sizes with a patch...

  12. Hybrid surface platform for the simultaneous detection of proteins and DNA using a surface plasmon resonance (SPR) imaging sensor

    Czech Academy of Sciences Publication Activity Database

    Homola, Jiří; Piliarik, Marek; Ladd, J.; Taylor, A.; Shaoyi, J.

    2008-01-01

    Roč. 80, č. 11 (2008), s. 4231-4236 ISSN 0003-2700 R&D Projects: GA AV ČR KAN200670701 Institutional research plan: CEZ:AV0Z20670512 Keywords : Surface plasmon resonance imaging * DNA -directed immobilization * protein array Subject RIV: JB - Sensors, Measurment, Regulation Impact factor: 5.712, year: 2008

  13. Functionalized Polyacrylonitrile Nanofibrous Membranes for Covalent Immobilization of Glucose Oxidase.

    Science.gov (United States)

    Manuel, James; Kim, Miso; Dharela, Rohini; Chauhan, Ghanshyam S; Fapyane, Deby; Lee, Soo-Jin; Chang, In Seop; Kang, Seo-Hee; Kim, Seon-Won; Ahn, Jou-Hyeon

    2015-01-01

    Nanofibrous membrane (NFM) with uniform morphology and large surface area was prepared from 10% solution of polyacrylonitrile (PAN) in N,N-dimethylformamide by electrospinning technique. NFM was chemically modified for use as a support for the immobilization of glucose oxidase. Chemical modification of NFM was carried out by two different methods. In the first method, the cyano groups of PAN were modified to amino groups by a two-step process, while in the second method the carboxylic groups were generated first and then further reacted with hexamethylene diamine to create a reactive spacer arm for the immobilization of enzyme. Scanning electron microscopy studies showed that the surface morphology of NFM was not changed by chemical modification and its mechanical strength was improved. The immobilized glucose oxidase (GOx) retained 54 and 60% of its original activity up to 25 cycles with the PAN NFMs modified by the first and the second method, respectively. The GOx-immobilized NFM from the second method showed promising performance with higher enzyme immobilization, activity retention, and favorable kinetic parameters.

  14. The dependence of the nuclear charge form factor on short range correlations and surface fluctuation effects

    International Nuclear Information System (INIS)

    Massen, S. E.; Garistov, V. P.; Grypeos, M. E.

    1996-01-01

    The effects of nuclear surface fluctuations on harmonic oscillator elastic charge form factor of light nuclei are investigated, simultaneously approximating the short-range correlations through a Jastrow correlation factor. Inclusion of the surface fluctuation effects within this description, by truncating the cluster expansion at the two-body part, is found to improve somewhat the fit to the elastic charge form-factor of 16 O and 40 Ca. However, the convergence of the cluster expansion is expected to deteriorate. An additional finding is that surface-fluctuation correlations produce a drastic change in the asymptotic behaviour of the point-proton form-factor, which now falls off quite slowly (i.e. as const.q -4 ) at large values of the momentum transfer q

  15. Combinational Effect of Cell Adhesion Biomolecules and Their Immobilized Polymer Property to Enhance Cell-Selective Adhesion

    Directory of Open Access Journals (Sweden)

    Rio Kurimoto

    2016-01-01

    Full Text Available Although surface immobilization of medical devices with bioactive molecules is one of the most widely used strategies to improve biocompatibility, the physicochemical properties of the biomaterials significantly impact the activity of the immobilized molecules. Herein we investigate the combinational effects of cell-selective biomolecules and the hydrophobicity/hydrophilicity of the polymeric substrate on selective adhesion of endothelial cells (ECs, fibroblasts (FBs, and smooth muscle cells (SMCs. To control the polymeric substrate, biomolecules are immobilized on thermoresponsive poly(N-isopropylacrylamide-co-2-carboxyisopropylacrylamide (poly(NIPAAm-co-CIPAAm-grafted glass surfaces. By switching the molecular conformation of the biomolecule-immobilized polymers, the cell-selective adhesion performances are evaluated. In case of RGDS (Arg-Gly-Asp-Ser peptide-immobilized surfaces, all cell types adhere well regardless of the surface hydrophobicity. On the other hand, a tri-Arg-immobilized surface exhibits FB-selectivity when the surface is hydrophilic. Additionally, a tri-Ile-immobilized surface exhibits EC-selective cell adhesion when the surface is hydrophobic. We believe that the proposed concept, which is used to investigate the biomolecule-immobilized surface combination, is important to produce new biomaterials, which are highly demanded for medical implants and tissue engineering.

  16. An orientation analysis method for protein immobilized on quantum dot particles

    Energy Technology Data Exchange (ETDEWEB)

    Aoyagi, Satoka, E-mail: aoyagi@life.shimane-u.ac.jp [Faculty of Life and Environmental Science, Shimane University, 1060 Matsue-shi, Shimane 690-8504 (Japan); Inoue, Masae [Toyota Central R and D Labs., Inc., Nagakute, Aichi 480-1192 (Japan)

    2009-11-30

    The evaluation of orientation of biomolecules immobilized on nanodevices is crucial for the development of high performance devices. Such analysis requires ultra high sensitivity so as to be able to detect less than one molecular layer on a device. Time-of-flight secondary ion mass spectrometry (TOF-SIMS) has sufficient sensitivity to evaluate the uppermost surface structure of a single molecular layer. The objective of this study is to develop an orientation analysis method for proteins immobilized on nanomaterials such as quantum dot particles, and to evaluate the orientation of streptavidin immobilized on quantum dot particles by means of TOF-SIMS. In order to detect fragment ions specific to the protein surface, a monoatomic primary ion source (Ga{sup +}) and a cluster ion source (Au{sub 3}{sup +}) were employed. Streptavidin-immobilized quantum dot particles were immobilized on aminosilanized ITO glass plates at amino groups by covalent bonding. The reference samples streptavidin directly immobilized on ITO plates were also prepared. All samples were dried with a freeze dryer before TOF-SIMS measurement. The positive secondary ion spectra of each sample were obtained using TOF-SIMS with Ga{sup +} and Au{sub 3}{sup +}, respectively, and then they were compared so as to characterize each sample and detect the surface structure of the streptavidin immobilized with the biotin-immobilized quantum dots. The chemical structures of the upper surface of the streptavidin molecules immobilized on the quantum dot particles were evaluated with TOF-SIMS spectra analysis. The indicated surface side of the streptavidin molecules immobilized on the quantum dots includes the biotin binding site.

  17. An orientation analysis method for protein immobilized on quantum dot particles

    International Nuclear Information System (INIS)

    Aoyagi, Satoka; Inoue, Masae

    2009-01-01

    The evaluation of orientation of biomolecules immobilized on nanodevices is crucial for the development of high performance devices. Such analysis requires ultra high sensitivity so as to be able to detect less than one molecular layer on a device. Time-of-flight secondary ion mass spectrometry (TOF-SIMS) has sufficient sensitivity to evaluate the uppermost surface structure of a single molecular layer. The objective of this study is to develop an orientation analysis method for proteins immobilized on nanomaterials such as quantum dot particles, and to evaluate the orientation of streptavidin immobilized on quantum dot particles by means of TOF-SIMS. In order to detect fragment ions specific to the protein surface, a monoatomic primary ion source (Ga + ) and a cluster ion source (Au 3 + ) were employed. Streptavidin-immobilized quantum dot particles were immobilized on aminosilanized ITO glass plates at amino groups by covalent bonding. The reference samples streptavidin directly immobilized on ITO plates were also prepared. All samples were dried with a freeze dryer before TOF-SIMS measurement. The positive secondary ion spectra of each sample were obtained using TOF-SIMS with Ga + and Au 3 + , respectively, and then they were compared so as to characterize each sample and detect the surface structure of the streptavidin immobilized with the biotin-immobilized quantum dots. The chemical structures of the upper surface of the streptavidin molecules immobilized on the quantum dot particles were evaluated with TOF-SIMS spectra analysis. The indicated surface side of the streptavidin molecules immobilized on the quantum dots includes the biotin binding site.

  18. SU-E-T-577: Obliquity Factor and Surface Dose in Proton Beam Therapy

    International Nuclear Information System (INIS)

    Das, I; Andersen, A; Coutinho, L

    2015-01-01

    Purpose: The advantage of lower skin dose in proton beam may be diminished creating radiation related sequalae usually seen with photon and electron beams. This study evaluates the surface dose as a complex function of beam parameters but more importantly the effect of beam angle. Methods: Surface dose in proton beam depends on the beam energy, source to surface distance, the air gap between snout and surface, field size, material thickness in front of surface, atomic number of the medium, beam angle and type of nozzle (ie double scattering, (DS), uniform scanning (US) or pencil beam scanning (PBS). Obliquity factor (OF) is defined as ratio of surface dose in 0° to beam angle Θ. Measurements were made in water phantom at various beam angles using very small microdiamond that has shown favorable beam characteristics for high, medium and low proton energy. Depth dose measurements were performed in the central axis of the beam in each respective gantry angle. Results: It is observed that surface dose is energy dependent but more predominantly on the SOBP. It is found that as SSD increases, surface dose decreases. In general, SSD, and air gap has limited impact in clinical proton range. High energy has higher surface dose and so the beam angle. The OF rises with beam angle. Compared to OF of 1.0 at 0° beam angle, the value is 1.5, 1.6, 1,7 for small, medium and large range respectively for 60 degree angle. Conclusion: It is advised that just like range and SOBP, surface dose should be clearly understood and a method to reduce the surface dose should be employed. Obliquity factor is a critical parameter that should be accounted in proton beam therapy and a perpendicular beam should be used to reduce surface dose

  19. Optimizing Immobilized Enzyme Performance in Cell-Free Environments to Produce Liquid Fuels

    Energy Technology Data Exchange (ETDEWEB)

    Belfort, Georges [Rensselaer Polytechnic Inst., Troy, NY (United States). Dept. of Chemical and Biological Engineering; Grimaldi, Joseph J. [Rensselaer Polytechnic Inst., Troy, NY (United States). Dept. of Chemical and Biological Engineering

    2015-01-27

    Limitations on biofuel production using cell culture (Escherichia coli, Clostridium, Saccharomyces cerevisiae, brown microalgae, blue-green algae and others) include low product (alcohol) concentrations (≤0.2 vol%) due to feedback inhibition, instability of cells, and lack of economical product recovery processes. To overcome these challenges, an alternate simplified biofuel production scheme was tested based on a cell-free immobilized enzyme system. Using this cell free system, we were able to obtain about 2.6 times higher concentrations of iso-butanol using our non-optimized system as compared with live cell systems. This process involved two steps: (i) converts acid to aldehyde using keto-acid decarboxylase (KdcA), and (ii) produces alcohol from aldehyde using alcohol dehydrogenase (ADH) with a cofactor (NADH) conversion from inexpensive formate using a third enzyme, formate dehydrogenase (FDH). To increase stability and conversion efficiency with easy separations, the first two enzymes were immobilized onto methacrylate resin. Fusion proteins of labile KdcA (fKdcA) were expressed to stabilize the covalently immobilized KdcA. Covalently immobilized ADH exhibited long-term stability and efficient conversion of aldehyde to alcohol over multiple batch cycles without fusions. High conversion rates and low protein leaching were achieved by covalent immobilization of enzymes on methacrylate resin. The complete reaction scheme was demonstrated by immobilizing both ADH and fKdcA and using FDH free in solution. The new system without in situ removal of isobutanol achieved a 55% conversion of ketoisovaleric acid to isobutanol at a concentration of 0.5 % (v/v). Further increases in titer will require continuous removal of the isobutanol using our novel brush membrane system that exhibits a 1.5 fold increase in the separation factor of isobutanol from water versus that obtained for commercial silicone rubber membranes. These bio-inspired brush membranes are based on the

  20. Calculated dose factors for the radiosensitive tissues in bone irradiated by surface-deposited radionuclides

    International Nuclear Information System (INIS)

    Spiers, F.W.; Whitwell, J.R.; Beddoe, A.H.

    1978-01-01

    The method of calculating dose factors for the haemopoietic marrow and endosteal tissues in human trabecular bone, used by Whitwell and Spiers for volume-seeking radionuclides, has been developed for the case of radionuclides which are deposited as very thin layers on bone surfaces. The Monte Carlo method is again used, but modifications to the computer program are made to allow for a surface rather than a volume source of particle emission. The principal change is the introduction of a surface-orientation factor which is shown to have a value of approximately 2, varying slightly with bone structure. Results are given for β-emitting radionuclides ranging from 171 Tm(anti Esub(β) = 0.025 MeV) to 90 Y(anti Esub(β) = 0.93 MeV), and also for the α-emitter 239 Pu. It is shown that where the particle ranges are short compared with the dimensions of the bone structures the dose factors for the surface seekers are much greater than those for the volume seekers. For long range particles the dose factors for surface- and volume-seeking radionuclides converge. Comparisons are given relating the dose factors calculated in this paper on the basis of measured bone structures to those of other workers based on single plane geometry. (author)

  1. Status of plutonium ceramic immobilization processes and immobilization forms

    International Nuclear Information System (INIS)

    Ebbinghaus, B.B.; Van Konynenburg, R.A.; Vance, E.R.; Jostsons, A.

    1996-01-01

    Immobilization in a ceramic followed by permanent emplacement in a repository or borehole is one of the alternatives currently being considered by the Fissile Materials Disposition Program for the ultimate disposal of excess weapons-grade plutonium. To make Pu recovery more difficult, radioactive cesium may also be incorporated into the immobilization form. Valuable data are already available for ceramics form R ampersand D efforts to immobilize high-level and mixed wastes. Ceramics have a high capacity for actinides, cesium, and some neutron absorbers. A unique characteristic of ceramics is the existence of mineral analogues found in nature that have demonstrated actinide immobilization over geologic time periods. The ceramic form currently being considered for plutonium disposition is a synthetic rock (SYNROC) material composed primarily of zirconolite (CaZrTi 2 O 7 ), the desired actinide host phase, with lesser amounts of hollandite (BaAl 2 Ti 6 O 16 ) and rutile (TiO 2 ). Alternative actinide host phases are also being considered. These include pyrochlore (Gd 2 Ti 2 O 7 ), zircon (ZrSiO 4 ), and monazite (CePO 4 ), to name a few of the most promising. R ampersand D activities to address important technical issues are discussed. Primarily these include moderate scale hot press fabrications with plutonium, direct loading of PuO 2 powder, cold press and sinter fabrication methods, and immobilization form formulation issues

  2. Performance Enhancements to the Hanford Waste Treatment and Immobilization Plant Low-Activity Waste Vitrification System

    International Nuclear Information System (INIS)

    Hamel, W. F.; Gerdes, K.; Holton, L. K.; Pegg, I.L.; Bowan, B.W.

    2006-01-01

    The U.S Department of Energy Office of River Protection (DOE-ORP) is constructing a Waste Treatment and Immobilization Plant (WTP) for the treatment and vitrification of underground tank wastes stored at the Hanford Site in Washington State. The WTP comprises four major facilities: a pretreatment facility to separate the tank waste into high level waste (HLW) and low-activity waste (LAW) process streams, a HLW vitrification facility to immobilize the HLW fraction; a LAW vitrification facility to immobilize the LAW fraction, and an analytical laboratory to support the operations of all four treatment facilities. DOE has established strategic objectives to optimize the performance of the WTP facilities and the LAW and HLW waste forms to reduce the overall schedule and cost for treatment and vitrification of the Hanford tank wastes. This strategy has been implemented by establishing performance expectations in the WTP contract for the facilities and waste forms. In addition, DOE, as owner-operator of the WTP facilities, continues to evaluate 1) the design, to determine the potential for performance above the requirements specified in the WTP contract; and 2) improvements in production of the LAW and HLW waste forms. This paper reports recent progress directed at improving production of the LAW waste form. DOE's initial assessment, which is based on the work reported in this paper, is that the treatment rate of the WTP LAW vitrification facility can be increased by a factor of 2 to 4 with a combination of revised glass formulations, modest increases in melter glass operating temperatures, and a second-generation LAW melter with a larger surface area. Implementing these improvements in the LAW waste immobilization capability can benefit the LAW treatment mission by reducing the cost of waste treatment. (authors)

  3. Immobilization of different biomolecules by atomic force microscopy

    Directory of Open Access Journals (Sweden)

    Hölzel Ralph

    2010-05-01

    Full Text Available Abstract Background Micrometer resolution placement and immobilization of probe molecules is an important step in the preparation of biochips and a wide range of lab-on-chip systems. Most known methods for such a deposition of several different substances are costly and only suitable for a limited number of probes. In this article we present a flexible procedure for simultaneous spatially controlled immobilization of functional biomolecules by molecular ink lithography. Results For the bottom-up fabrication of surface bound nanostructures a universal method is presented that allows the immobilization of different types of biomolecules with micrometer resolution. A supporting surface is biotinylated and streptavidin molecules are deposited with an AFM (atomic force microscope tip at distinct positions. Subsequent incubation with a biotinylated molecule species leads to binding only at these positions. After washing streptavidin is deposited a second time with the same AFM tip and then a second biotinylated molecule species is coupled by incubation. This procedure can be repeated several times. Here we show how to immobilize different types of biomolecules in an arbitrary arrangement whereas most common methods can deposit only one type of molecules. The presented method works on transparent as well as on opaque substrates. The spatial resolution is better than 400 nm and is limited only by the AFM's positional accuracy after repeated z-cycles since all steps are performed in situ without moving the supporting surface. The principle is demonstrated by hybridization to different immobilized DNA oligomers and was validated by fluorescence microscopy. Conclusions The immobilization of different types of biomolecules in high-density microarrays is a challenging task for biotechnology. The method presented here not only allows for the deposition of DNA at submicrometer resolution but also for proteins and other molecules of biological relevance that

  4. Modification of the ASME code z-factor for circumferential surface crack in nuclear ferritic pipings

    International Nuclear Information System (INIS)

    Choi, Young Hwan; Chung, Yon Ki; Koh, Wan Young; Lee, Joung Bae

    1996-01-01

    The purpose of this paper is to modify the ASME Code Z-Factor, which is used in the evaluation of circumferential surface crack in nuclear ferritic pipings. The ASME Code Z-Factor is a load multiplier to compensate plastic load with elasto-plastic load. The current ASME Code Z-Factor underestimates pipe maximum load. In this study, the original SC. TNP method is modified first because the original SC. TNP method has a problem that the maximum allowable load predicted from the original SC. TNP method is slightly higher than that measured from the experiment. Then the new Z-Factor is developed using the modified SC. TNP method. The desirability of both the modified SC. TNP method and the new Z-Factor is examined using the experimental results for the circumferential surface crack in pipings. The results show that (1) the modified SC. TNP method is good for predicting the circumferential surface crack behavior in pipings, and (2) the Z-Factor obtained from the modified SC. TNP method well predicts the behavior of circumferential surface crack in ferritic pipings. 30 refs., 13 figs., 4 tabs. (author)

  5. Cyclic muscle twitch contraction inhibits immobilization-induced muscle contracture and fibrosis in rats.

    Science.gov (United States)

    Yoshimura, Ayana; Sakamoto, Junya; Honda, Yuichiro; Kataoka, Hideki; Nakano, Jiro; Okita, Minoru

    2017-09-01

    We investigated the effects of cyclic muscle twitch contraction caused by neuromuscular electrical stimulation (NMES) on immobilization-induced muscle contracture and fibrosis in rats. Twenty-nine rats were divided into control, immobilization, and immobilization with muscle contraction groups. The ankle joints of the immobilization and muscle contraction rats were fixed in full plantar flexion with a plaster cast for 4 weeks. In the muscle contraction group, cyclic muscle twitch contraction of the soleus muscle was induced using a commercial device (1 Hz, 4 ± 2 mA, 60 min/day, 5 times/week) with the ankle joint immobilized. The dorsiflexion range of ankle joint motion in the muscle contraction group was significantly greater than that in the immobilization group. The expressions of fibrosis-related genes (i.e., hypoxia inducible factor-1α, transforming growth factor-β1, α-smooth muscle actin, and types I and III collagen) were significantly decreased in the muscle contraction group compared to the immobilization group. The fluorescence intensities of type I and type III collagen in the perimysium and endomysium in the muscle contraction group were significantly decreased compared to the immobilization group. These results suggest that cyclic muscle twitch contraction induced by NMES might alleviate skeletal muscle fibrosis, reducing immobilization-induced muscle contracture.

  6. Immobilization of bioactive fibroblast growth factor-2 into cubic proteinous microcrystals (Bombyx mori cypovirus polyhedra) that are insoluble in a physiological cellular environment.

    Science.gov (United States)

    Mori, Hajime; Shukunami, Chisa; Furuyama, Akiko; Notsu, Hiroyuki; Nishizaki, Yuriko; Hiraki, Yuji

    2007-06-08

    The supramolecular architecture of the extracellular matrix and the disposition of its specific accessory molecules give rise to variable heterotopic signaling cues for single cells. Here we have described the successful occlusion of human fibroblast growth factor-2 (FGF-2) into the cubic inclusion bodies (FGF-2 polyhedra) of the Bombyx mori cytoplasmic polyhedrosis virus (BmCPV). The polyhedra are proteinous cubic crystals of several microns in size that are insoluble in the extracellular milieu. Purified FGF-2 polyhedra were found to stimulate proliferation and phosphorylation of p44/p42 mitogen-activated protein kinase in cultured fibroblasts. Moreover, cellular responses were blocked by a synthetic inhibitor of the FGF signaling pathway, SU5402, suggesting that FGF-2 polyhedra indeed act through FGF receptors. Furthermore, FGF-2 polyhedra retained potent growth stimulatory properties even after desiccation. We have demonstrated that BmCPV polyhedra microcrystals that occlude extracellular signaling proteins are a novel and versatile tool that can be employed to analyze cellular behavior at the single cell level.

  7. An experimental method to determine the electrostatic field enhancement factor of a practical conductor surface

    DEFF Research Database (Denmark)

    McAllister, Iain Wilson; Crichton, George C

    1989-01-01

    A method of determining the field enhancement factor of a practical conductor is presented. The method is developed from a modified theory of discharge onset in a gaseous medium. This modification incorporates the influence of conductor surface roughness. Onset data from an experimental study...... that utilized electrodes of varying surface roughness are examined, and the results obtained using the proposed method are discussed with reference to both the underlying theory and the practical aspects of the experimental measurements...

  8. Analysis of alternatives for immobilized low activity waste disposal

    Energy Technology Data Exchange (ETDEWEB)

    Burbank, D.A.

    1997-10-28

    This report presents a study of alternative disposal system architectures and implementation strategies to provide onsite near-surface disposal capacity to receive the immobilized low-activity waste produced by the private vendors. The analysis shows that a flexible unit strategy that provides a suite of design solutions tailored to the characteristics of the immobilized low-activity waste will provide a disposal system that best meets the program goals of reducing the environmental, health, and safety impacts; meeting the schedule milestones; and minimizing the life-cycle cost of the program.

  9. Analysis of alternatives for immobilized low-activity waste disposal

    International Nuclear Information System (INIS)

    Burbank, D.A.

    1997-01-01

    This report presents a study of alternative disposal system architectures and implementation strategies to provide onsite near-surface disposal capacity to receive the immobilized low-activity waste produced by the private vendors. The analysis shows that a flexible unit strategy that provides a suite of design solutions tailored to the characteristics of the immobilized low-activity waste will provide a disposal system that best meets the program goals of reducing the environmental, health, and safety impacts; meeting the schedule milestones; and minimizing the life-cycle cost of the program

  10. Magnetic bead cellulose as a suitable support for immobilization of .alpha.-chymotrypsin

    Czech Academy of Sciences Publication Activity Database

    Přikryl, P.; Lenfeld, Jiří; Horák, Daniel; Tichá, M.; Kučerová, Z.

    2012-01-01

    Roč. 168, č. 2 (2012), s. 295-305 ISSN 0273-2289 R&D Projects: GA ČR GA203/09/0857 Institutional research plan: CEZ:AV0Z40500505 Keywords : enzyme immobilization * human gastric juice * immobilized α-chymotrypsin Subject RIV: CB - Analytical Chemistry, Separation Impact factor: 1.893, year: 2012

  11. Immobilization needs and technology programs

    International Nuclear Information System (INIS)

    Gray, L.W.; Kan, T.; Shaw, H.; Armantrout, G.

    1995-01-01

    In the aftermath of the Cold War, the US and Russia agreed to large reductions in nuclear weapons. To aid in the selection of long-term management options, DOE has undertaken a multifaceted study to select options for storage and disposition of plutonium in keeping with US policy that plutonium must be subjected to the highest standards of safety, security, and accountability. One alternative being considered is immobilization. To arrive at a suitable immobilization form, we first reviewed published information on high-level waste immobilization technologies and identified 72 possible plutonium immobilization forms to be prescreened. Surviving forms were further screened using multi-attribute utility analysis to determine the most promising technology families. Promising immobilization families were further evaluated to identify chemical, engineering, environmental, safety, and health problems that remain to be solved prior to making technical decisions as to the viability of using the form for long- term disposition of plutonium. From this evaluation, a detailed research and development plan has been developed to provide answers to these remaining questions

  12. Binary polypeptide system for permanent and oriented protein immobilization

    Directory of Open Access Journals (Sweden)

    Bailes Julian

    2010-05-01

    Full Text Available Abstract Background Many techniques in molecular biology, clinical diagnostics and biotechnology rely on binary affinity tags. The existing tags are based on either small molecules (e.g., biotin/streptavidin or glutathione/GST or peptide tags (FLAG, Myc, HA, Strep-tag and His-tag. Among these, the biotin-streptavidin system is most popular due to the nearly irreversible interaction of biotin with the tetrameric protein, streptavidin. The major drawback of the stable biotin-streptavidin system, however, is that neither of the two tags can be added to a protein of interest via recombinant means (except for the Strep-tag case leading to the requirement for chemical coupling. Results Here we report a new immobilization system which utilizes two monomeric polypeptides which self-assemble to produce non-covalent yet nearly irreversible complex which is stable in strong detergents, chaotropic agents, as well as in acids and alkali. Our system is based on the core region of the tetra-helical bundle known as the SNARE (soluble N-ethylmaleimide-sensitive factor attachment protein receptor complex. This irreversible protein attachment system (IPAS uses either a shortened syntaxin helix and fused SNAP25-synaptobrevin or a fused syntaxin-synaptobrevin and SNAP25 allowing a two-component system suitable for recombinant protein tagging, capture and immobilization. We also show that IPAS is suitable for use with traditional beads and chromatography, planar surfaces and Biacore, gold nanoparticles and for protein-protein interaction in solution. Conclusions IPAS offers an alternative to chemical cross-linking, streptavidin-biotin system and to traditional peptide affinity tags and can be used for a wide range of applications in nanotechnology and molecular sciences.

  13. Binary polypeptide system for permanent and oriented protein immobilization.

    Science.gov (United States)

    Ferrari, Enrico; Darios, Frédéric; Zhang, Fan; Niranjan, Dhevahi; Bailes, Julian; Soloviev, Mikhail; Davletov, Bazbek

    2010-05-12

    Many techniques in molecular biology, clinical diagnostics and biotechnology rely on binary affinity tags. The existing tags are based on either small molecules (e.g., biotin/streptavidin or glutathione/GST) or peptide tags (FLAG, Myc, HA, Strep-tag and His-tag). Among these, the biotin-streptavidin system is most popular due to the nearly irreversible interaction of biotin with the tetrameric protein, streptavidin. The major drawback of the stable biotin-streptavidin system, however, is that neither of the two tags can be added to a protein of interest via recombinant means (except for the Strep-tag case) leading to the requirement for chemical coupling. Here we report a new immobilization system which utilizes two monomeric polypeptides which self-assemble to produce non-covalent yet nearly irreversible complex which is stable in strong detergents, chaotropic agents, as well as in acids and alkali. Our system is based on the core region of the tetra-helical bundle known as the SNARE (soluble N-ethylmaleimide-sensitive factor attachment protein receptor) complex. This irreversible protein attachment system (IPAS) uses either a shortened syntaxin helix and fused SNAP25-synaptobrevin or a fused syntaxin-synaptobrevin and SNAP25 allowing a two-component system suitable for recombinant protein tagging, capture and immobilization. We also show that IPAS is suitable for use with traditional beads and chromatography, planar surfaces and Biacore, gold nanoparticles and for protein-protein interaction in solution. IPAS offers an alternative to chemical cross-linking, streptavidin-biotin system and to traditional peptide affinity tags and can be used for a wide range of applications in nanotechnology and molecular sciences.

  14. The glycoprotein Ib-IX-V complex contributes to tissue factor-independent thrombin generation by recombinant factor VIIa on the activated platelet surface

    NARCIS (Netherlands)

    Weeterings, Cees; de Groot, Philip G.; Adelmeijer, Jelle; Lisman, Ton

    2008-01-01

    Several lines of evidence suggest that recombinant factor VIIa (rFVIIa) is able to activate factor X on an activated platelet, in a tissue factor-independent manner. We hypothesized that, besides the anionic surface, a receptor on the activated platelet surface is involved in this process. Here, we

  15. Approaching Immobilization of Enzymes onto Open Porous Basotect®

    Directory of Open Access Journals (Sweden)

    Peter J. Allertz

    2017-11-01

    Full Text Available For the first time, commercial macroporous melamine formaldehyde foam Basotect® (BT was used as a basic carrier material for both adsorptive and covalent enzyme immobilization. In order to access inherent amino groups, the Basotect® surface was pretreated with hydrochloric acid. The resulting material revealed 6 nmol of superficial amino groups per milligram Basotect®. Different optimized strategies for tethering the laccase from Trametes versicolor and the lipase from Thermomyces lanuginosus onto the pre-treated Basotect® surface were studied. Particularly, for covalent immobilization, two different strategies were pursued: lipase was tethered via a cross-linking method using 1-ethyl-3-(3-dimethylaminopropylcarbodiimide, and laccase was bound after functionalizing Basotect® with hydrophilic copolymer poly(ethylene-alt-maleic anhydride (PEMA. Prior to laccase immobilization, the PEMA coating of Basotect® was verified by ATR-FTIR analysis. Subsequent quantification of available high-reactive PEMA anhydride moieties revealed an amount of 1028 ± 73 nmol per mg Basotect®. The surface-bound enzyme amounts were quantified as 4.1–5.8 μg per mg Basotect®. A theoretical surface-covered enzyme mass for the ideal case that an enzyme monolayer was immobilized onto the Basotect® surface was calculated and compared to the amount of adsorptive and covalently bound enzymes before and after treatment with SDS. Furthermore, the enzyme activities were determined for the different immobilization approaches, and the stability during storage over time and against sodium dodecyl sulfate treatment was monitored. Additionally, PEMA-BT-bound laccase was tested for the elimination of anthropogenic micropollutant bisphenol A from contaminated water in a cost-effective and environmentally-friendly way and resulted in a degradation rate higher than 80%.

  16. Enhanced bone morphogenetic protein-2 performance on hydroxyapatite ceramic surfaces.

    Science.gov (United States)

    Schuessele, A; Mayr, H; Tessmar, J; Goepferich, A

    2009-09-15

    The immobilization of biomolecules on biomaterial surfaces allows for the control of their localization and retention. In numerous studies, proteins have been simply adsorbed to enhance the biological performance of various materials in vivo. We investigated the potential of surface modification techniques on hydroxyapatite (HA) ceramic discs in an in vitro approach. A novel method for protein immobilization was evaluated using the aminobisphosphonates pamidronate and alendronate, which are strong Ca chelating agents, and was compared with the established silanization technique. Lysozyme and bone morphogenetic protein-2 (BMP-2) were used to assess the suitability of the two surface modification methods with regard to the enzymatic activity of lysozyme and to the capacity of BMP-2 to stimulate the osteoblastic differentiation of C2C12 mouse myoblasts. After immobilization, a 2.5-fold increase in enzymatic activity of lysozyme was observed compared with the control. The alkaline phosphatase activity per cell stimulated by immobilized BMP-2 was 2.5-fold higher [9 x 10(-6) I.U.] than the growth factor on unmodified surfaces [2-4 x 10(-6) I.U.]. With regard to the increase in protein activity, both procedures lead to equivalent results. Thus, the bisphosphonate-based surface modification represents a safe and easy alternative for the attachment of proteins to HA surfaces. Copyright 2008 Wiley Periodicals, Inc.

  17. Stabilization of enzymes by multipoint covalent immobilization on supports activated with glyoxyl groups.

    Science.gov (United States)

    López-Gallego, Fernando; Fernandez-Lorente, Gloria; Rocha-Martin, Javier; Bolivar, Juan M; Mateo, Cesar; Guisan, Jose M

    2013-01-01

    Stabilization of enzymes via immobilization techniques is a valuable approach in order to convert a necessary protocol (immobilization) into a very interesting tool to improve key enzyme properties (stabilization). Multipoint covalent attachment of each immobilized enzyme molecule may promote a very interesting stabilizing effect. The relative distances among all enzyme residues involved in immobilization has to remain unaltered during any conformational change induced by any distorting agent. Amino groups are very interesting nucleophiles placed on protein surfaces. The immobilization of enzyme through the region having the highest amount of amino groups (Lys residues) is key for a successful stabilization. Glyoxyl groups are small aliphatic aldehydes that form very unstable Schiff's bases with amino groups and they do not seem to be useful for enzyme immobilization at neutral pH. However, under alkaline conditions, glyoxyl supports are able to immobilize enzymes via a first multipoint covalent immobilization through the region having the highest amount of Lysine groups. Activation of supports with a high surface density of glyoxyl groups and the performance of very intense enzyme-support multipoint covalent attachments are here described.

  18. Immobilization of α-amylase onto poly(glycidyl methacrylate) grafted electrospun fibers by ATRP

    International Nuclear Information System (INIS)

    Oktay, Burcu; Demir, Serap; Kayaman-Apohan, Nilhan

    2015-01-01

    In this study, novel α-amylase immobilized poly(vinyl alcohol) (PVA) nanofibers were prepared. The PVA nanofiber surfaces were functionalized with 2-bromoisobutyryl bromide (BiBBr) and followed by surface initiated atom transfer radical polymerization (SI-ATRP) of glycidyl methacrylate (GMA). The morphology of the poly(glycidyl methacrylate) (PGMA) grafted PVA nanofibers was characterized by scanning electron microscopy (SEM). Also PGMA brushes were confirmed by X-ray photo electron microscopy (XPS). α-Amylase was immobilized in a one step process onto the PGMA grafted PVA nanofiber. The characteristic properties of the immobilized and free enzymes were examined. The thermal stability of the enzyme was improved and showed maximum activity at 37 °C by immobilization. pH values of the maximum activity of the free and immobilized enzymes were also found at 6.0 and 6.5, respectively. Free enzyme lost its activity completely within 15 days. The immobilized enzyme lost only 23.8% of its activity within 30 days. - Highlights: • Electrospun photocrosslinkable PVA nanofiber was prepared. • PGMA brushes were conducted on PVA nanofiber via SI-ATRP. • The immobilized enzyme showed maximum activity at pH 6.0 and at 37 °C. • Functionalized nanofibers may be used as promising supports for enzyme immobilization

  19. Immobilization of α-amylase onto poly(glycidyl methacrylate) grafted electrospun fibers by ATRP

    Energy Technology Data Exchange (ETDEWEB)

    Oktay, Burcu; Demir, Serap; Kayaman-Apohan, Nilhan, E-mail: napohan@marmara.edu.tr

    2015-05-01

    In this study, novel α-amylase immobilized poly(vinyl alcohol) (PVA) nanofibers were prepared. The PVA nanofiber surfaces were functionalized with 2-bromoisobutyryl bromide (BiBBr) and followed by surface initiated atom transfer radical polymerization (SI-ATRP) of glycidyl methacrylate (GMA). The morphology of the poly(glycidyl methacrylate) (PGMA) grafted PVA nanofibers was characterized by scanning electron microscopy (SEM). Also PGMA brushes were confirmed by X-ray photo electron microscopy (XPS). α-Amylase was immobilized in a one step process onto the PGMA grafted PVA nanofiber. The characteristic properties of the immobilized and free enzymes were examined. The thermal stability of the enzyme was improved and showed maximum activity at 37 °C by immobilization. pH values of the maximum activity of the free and immobilized enzymes were also found at 6.0 and 6.5, respectively. Free enzyme lost its activity completely within 15 days. The immobilized enzyme lost only 23.8% of its activity within 30 days. - Highlights: • Electrospun photocrosslinkable PVA nanofiber was prepared. • PGMA brushes were conducted on PVA nanofiber via SI-ATRP. • The immobilized enzyme showed maximum activity at pH 6.0 and at 37 °C. • Functionalized nanofibers may be used as promising supports for enzyme immobilization.

  20. Immobilization and controlled release of drug using plasma polymerized thin film

    Energy Technology Data Exchange (ETDEWEB)

    Myung, Sung-Woon [Department of Dental Materials, School of Dentistry, MRC Center, Chosun University, 309 Pilmun-daero, Dong-gu, Gwangju (Korea, Republic of); Jung, Sang-Chul [Department of Environmental Engineering, Sunchon National University, Sunchon 540-742 (Korea, Republic of); Kim, Byung-Hoon, E-mail: kim5055@chosun.ac.kr [Department of Dental Materials, School of Dentistry, MRC Center, Chosun University, 309 Pilmun-daero, Dong-gu, Gwangju (Korea, Republic of)

    2015-06-01

    In this study, plasma polymerization of acrylic acid was employed to immobilize drug and control its release. Doxorubicin (DOX) was immobilized covalently on the glass surface deposited with plasma polymerized acrylic acid (PPAAc) thin film containing the carboxylic group. At first, the PPAAc thin film was coated on a glass surface at a pressure of 1.33 Pa and radio frequency (RF) discharge power of 20 W for 10 min. DOX was immobilized on the PPAAc deposition in a two environment of phosphate buffer saline (PBS) and dimethyl sulfoxide (DMSO) solutions. The DOX immobilized surface was characterized by scanning electron microscope, atomic force microscope and attenuated total reflection Fourier transform infrared spectroscopy. The DOX molecules were more immobilized in PBS than DMSO solution. The different immobilization and release profiles of DOX result from the solubility of hydrophobic DOX in aqueous and organic solutions. Second, in order to control the release of the drug, PPAAc thin film was covered over DOX dispersed layer. Different thicknesses and cross-linked PPAAc thin films by adjusting deposition time and RF discharge power were covered on the DOX layer dispersed. PPAAc thin film coated DOX layer reduced the release rate of DOX. The thickness control of plasma deposition allows controlling the release rate of drug. - Highlights: • Doxorubicin was immobilized on the surface of plasma polymerized acrylic acid thin film. • Release profile of doxorubicin was affected by aqueous and organic solutions. • Plasma polymerized acrylic acid thin film can be used to achieve controlled release.

  1. Immobilization and controlled release of drug using plasma polymerized thin film

    International Nuclear Information System (INIS)

    Myung, Sung-Woon; Jung, Sang-Chul; Kim, Byung-Hoon

    2015-01-01

    In this study, plasma polymerization of acrylic acid was employed to immobilize drug and control its release. Doxorubicin (DOX) was immobilized covalently on the glass surface deposited with plasma polymerized acrylic acid (PPAAc) thin film containing the carboxylic group. At first, the PPAAc thin film was coated on a glass surface at a pressure of 1.33 Pa and radio frequency (RF) discharge power of 20 W for 10 min. DOX was immobilized on the PPAAc deposition in a two environment of phosphate buffer saline (PBS) and dimethyl sulfoxide (DMSO) solutions. The DOX immobilized surface was characterized by scanning electron microscope, atomic force microscope and attenuated total reflection Fourier transform infrared spectroscopy. The DOX molecules were more immobilized in PBS than DMSO solution. The different immobilization and release profiles of DOX result from the solubility of hydrophobic DOX in aqueous and organic solutions. Second, in order to control the release of the drug, PPAAc thin film was covered over DOX dispersed layer. Different thicknesses and cross-linked PPAAc thin films by adjusting deposition time and RF discharge power were covered on the DOX layer dispersed. PPAAc thin film coated DOX layer reduced the release rate of DOX. The thickness control of plasma deposition allows controlling the release rate of drug. - Highlights: • Doxorubicin was immobilized on the surface of plasma polymerized acrylic acid thin film. • Release profile of doxorubicin was affected by aqueous and organic solutions. • Plasma polymerized acrylic acid thin film can be used to achieve controlled release

  2. Immobile Complex Verbs in Germanic

    DEFF Research Database (Denmark)

    Vikner, Sten

    2005-01-01

    Certain complex verbs in Dutch, German, and Swiss German do not undergo verb movement. The suggestion to be made in this article is that these ‘‘immobile'' verbs have to fulfill both the requirements imposed on complex verbs of the V° type (=verbs with non-separable prefixes) and the requirements...... imposed on complex verbs of the V* type (=verbs with separable prefixes). This results in such verbs being morphologically unexceptional, i.e., having a full set of forms but syntactically peculiar (‘‘immobile''), i.e., they can only occur in their base position. Any movement is incompatible with either...... frequent type with only one prefix-like part (e.g., German bauchreden/Dutch buikspreken ‘to ventriloquize'). This analysis will try to account not only for the data discussed in Haider (1993) and Koopman (1995) but also for the following: - why immobile verbs include verbs with only one prefix-like part...

  3. Synthesis of a polyhistidine-bearing amphipol and its use for immobilizing membrane proteins

    DEFF Research Database (Denmark)

    Giusti, Fabrice; Kessler, Pascal; Hansen, Randi Westh

    2015-01-01

    APol, are described. Its ability to immobilize MPs on nickel ion-bearing surfaces was tested using two complementary methods, immobilized metal affinity chromatography (IMAC) and surface plasmon resonance (SPR). Compared to a single His6-tag fused at one extremity of a MP, the presence of several His6-tags carried...... by the APol belt surrounding the transmembrane domain of a MP increases remarkably the affinity of the protein/APol complex for nickel ion-bearing SPR chips, whereas it does not show such a strong effect on an IMAC resin. HistAPol-mediated immobilization, which allows reversibility of the interaction and easy...

  4. Surface Water Quality Assessment and Prioritize the Factors Pollute This Water Using Topsis Fuzzy Hierarchical Analysis

    Directory of Open Access Journals (Sweden)

    Mehdi Komasi

    2017-03-01

    Full Text Available Background & Objective: Nowadays, according to growth of industry and increasing population, water resources are seriousely shortened. This lack of water resources will require special management to be considered in industry and agriculture. Among the various sources of water, surface waters are more susceptible to infection. The most important of these sources of pollution are industrial pollution, detergent, pesticides, radioactive materials, heat and salt concentration.  Materials & methods: In this article, at first the importance of each pollutant will be evaluated base on the effects and its results and then quality evaluation of surface water will be studied. In order to assess the relative importance of these pollutants primarily using TOPSIS software, prioritize these factors as one of the hierarchical analysis and then is modeled with decision tree method using Weka software, the importance of each factor is evaluated and if it does not meet the minimal importance of the decision tree will be removed. Results: The results obtained from the Topsis fuzzy analysis indicate that surface water and groundwater are exposed to pollution about 74% and 26% respectively among the six pollutants examined in this study. In addition, results obtaned from the hierarchical tree in software Weka has shown that the heat factor, soluble salts and industrial pollutants give impac factor or purity about 0.1338, 0.0523 and 1.2694 respectively. Conclusion: Surface water is at greater risk of being polluted compared with groundwater. The heat factor and low concentration of dissolved salts have the low impact and industrial pollutants are considered as the most influential factors in surface water pollution.

  5. Improvement of the homogeneity of protein-imprinted polymer films by orientated immobilization of the template

    International Nuclear Information System (INIS)

    Liu Lijian; Zheng Jingjing; Fang Guijie; Xie Weihong

    2012-01-01

    Highlights: ► MPH was genetically modified at its C-terminal with (Gly-Ser) 5 –Cys. ► MPH-L was immobilized with fixed orientation via disulfide chemistry. ► The immobilized MPH-L retained the activity of MPH. ► MPH-L formed a homogeneous template. ► Homogeneous MIP film was obtained with orientated immobilization of the template. - Abstract: A method for preparing homogeneous protein-imprinted polymer films with orientated immobilization of template is described. The template methyl parathion hydrolase (MPH) was modified with a peptide linker (Gly-Ser) 5 –Cys and was immobilized on a cover glass with a fixed orientation via the linker. The activity of the fusion enzyme (MPH-L) was evaluated by determining the product's absorbance at 405 nm (A 405 ). Both the free and the immobilized MPH-L showed higher retention of the bioactivity than the wide type enzyme (MPH-W) as revealed by the A 405 values for MPH-L free /MPH-W free (1.159/1.111) and for MPH-L immobilized /MPH-W immobilized (0.348/0.118). The immobilized MPH-L also formed a more homogeneous template stamp compared to the immobilized MPH-W. The molecularly imprinted polymer films prepared with the immobilized MPH-L exhibited high homogeneity with low Std. Deviations of 80 and 200 from the CL intensity mean volumes which were observed for batch-prepared films and an individual film, respectively. MPH-L-imprinted polymer film also had a larger template binding capacity indicated by higher CL intensity mean volume of 3900 INT over 2500 INT for MPH-W-imprinted films. The imprinted film prepared with the orientated immobilization of template showed an imprinting factor of 1.7, while the controls did not show an imprinting effect.

  6. Factors influencing prokaryotic community structure composition in sub-surface coastal sediments

    Science.gov (United States)

    Molari, Massimiliano; Giovannelli, Donato; d'Errico, Giuseppe; Manini, Elena

    2012-01-01

    Despite the major influence of the marine sub-surface area in carbon cycling and global biogeochemistry, there is little known of prokaryotic distribution and community structure information of the marine sub-surface and the factors that influence sub-surface prokaryotic assemblages. We provide quantitative estimations of active Bacteria and Archaea down vertical profiles of sub-surface coastal sediments from the southern Adriatic Sea (Manfredonia Gulf). Prokaryotic biomass, carbon incorporation and the metabolically active fraction were compared with environmental, trophic, and predatory variables down 100-cm sediment cores sectioned into 21 layers. Multiple regression analysis was used to identify variables that can explain the variance of community compositions down vertical profiles. CARD-FISH analysis showed Bacteria dominance for the first 11 cm below sediment surface. The community then changed significantly at increasing depth, towards Archaea dominance. The models tested show that prokaryotic abundance in superficial marine sediments is controlled by organic trophic resources, while in sub-surface sediments, active prokaryotic abundance is driven by environmental factors and predatory pressure, suggesting that the shift in prokaryotic community structure could be coupled to a change in life-style of microbial assemblages.

  7. Quality-factor and reflection-coefficient estimation using reflected surface waves

    NARCIS (Netherlands)

    Draganov, D.S.; Ruigrok, E.N.; Ghose, R.; Mikesell, D.; Van Wijk, K.

    2014-01-01

    We propose a method for estimating the reflection coefficient of a subvertical boundary and the the quality factor of the medium between a receiver and the subvertical boundary. The method uses surface waves from transient deterministic sources and is inspired by the occurrence of non-physical

  8. Basement to surface expressions and critical factors in the genesis of unconformity-related deposits

    International Nuclear Information System (INIS)

    Potter, Eric

    2014-01-01

    Two subprojects: 1) Basement to surface expressions of deep mineralization and refinement of critical factors leading to the genesis of unconformity-related uranium deposits; and 2) Recognition of uranium ore system alteration signatures in complex terranes: IOCG vs albite-hosted uranium vs volcanic-hosted uranium.

  9. Dimension of surface microstructure as an osteogenic factor in calcium phosphate ceramics

    NARCIS (Netherlands)

    Zhang, Jingwei; Luo, Xiaoman; Barbieri, D.; Barradas, A.M.C.; de Bruijn, Joost Dick; van Blitterswijk, Clemens; Yuan, Huipin

    2014-01-01

    Microporosity of calcium phosphate (CaP) ceramics has shown its essential role in osteoinduction by CaP ceramics after ectopic implantation. Here we show that not the microporosity but the dimension of surface microstructural features is likely the osteogenic factor. Two tricalcium phosphate

  10. Shielding Factors for Gamma Radiation from Activity Deposited on Structures and Ground Surfaces

    DEFF Research Database (Denmark)

    Jensen, Per Hedemann

    1985-01-01

    A computer model DEPSHIELD for the calculation of shielding factors for gamma radiation at indoor residences in multistorey and single-family houses has been developed. The model is based on the exponential point kernel that links the radiation flux density at a given detector point to a point......-source strength. The radiation sources considered in the model are fallout radioactivity deposited on roofs, outer walls, and ground surfaces. For any combination of source strength on roof, outer wall, and ground surface, the model calculates shielding factors for specified photon energies. The input data...... it possible to determine the dose reduction effect from a decontamination of the different surfaces. The model has been used in a study of the consequences of land contamination of Danish territory after hypothetical core-melt accidents at the Barseback nuclear power plant in Sweden. The model has also been...

  11. Influence of climatic factors and the ground surface on the required noise abatement from power equipment

    Science.gov (United States)

    Tupov, V. B.; Taratorin, A. A.

    2013-07-01

    The influence of climatic factors and the ground surface on the required noise abatement from the power equipment is analyzed. It is shown that annual oscillations of temperature and humidity lead to substantial variations in the levels of the sound and the sound pressure from the same source in the design point, while the ground effect surface can in some cases cause an increase in the sound pressure levels in the design point, and in other cases—their decrease. When developing the measures on sound suppression of the power equipment, it is recommended to take into account the influence of climatic factors depending on annual variations in temperature and humidity for this terrain as well as on the category of the ground surface.

  12. Acetylcholinesterase immobilization and characterization, and comparison of the activity of the porous silicon-immobilized enzyme with its free counterpart

    Science.gov (United States)

    Saleem, Muhammad; Rafiq, Muhammad; Seo, Sung-Yum; Lee, Ki Hwan

    2016-01-01

    A successful prescription is presented for acetylcholinesterase physically adsorbed on to a mesoporous silicon surface, with a promising hydrolytic response towards acetylthiocholine iodide. The catalytic behaviour of the immobilized enzyme was assessed by spectrophotometric bioassay using neostigmine methyl sulfate as a standard acetycholinesterase inhibitor. The surface modification was studied through field emission SEM, Fourier transform IR spectroscopy, energy-dispersive X-ray spectroscopy, cathode luminescence and X-ray photoelectron spectroscopy analysis, photoluminescence measurement and spectrophotometric bioassay. The porous silicon-immobilized enzyme not only yielded greater enzyme stability, but also significantly improved the native photoluminescence at room temperature of the bare porous silicon architecture. The results indicated the promising catalytic behaviour of immobilized enzyme compared with that of its free counterpart, with a greater stability, and that it aided reusability and easy separation from the reaction mixture. The porous silicon-immobilized enzyme was found to retain 50% of its activity, promising thermal stability up to 90°C, reusability for up to three cycles, pH stability over a broad pH of 4–9 and a shelf-life of 44 days, with an optimal hydrolytic response towards acetylthiocholine iodide at variable drug concentrations. On the basis of these findings, it was believed that the porous silicon-immobilized enzyme could be exploited as a reusable biocatalyst and for screening of acetylcholinesterase inhibitors from crude plant extracts and synthesized organic compounds. Moreover, the immobilized enzyme could offer a great deal as a viable biocatalyst in bioprocessing for the chemical and pharmaceutical industries, and bioremediation to enhance productivity and robustness. PMID:26839417

  13. Immobilization of alkaline phosphatase on magnetic particles by site-specific and covalent cross-linking catalyzed by microbial transglutaminase.

    Science.gov (United States)

    Moriyama, Kousuke; Sung, Kyunga; Goto, Masahiro; Kamiya, Noriho

    2011-06-01

    Bacterial alkaline phosphatase (BAP) was site-specifically and covalently immobilized on magnetic particles (MPs) using the enzymatic reaction of microbial transglutaminase (MTG). Immobilization efficiency was affected by the chemical surface treatment of MPs and immobilized BAP exhibited more than 90% of the initial activity after 10 rounds of recycling. Copyright © 2011 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  14. Downscaling Land Surface Temperature in Complex Regions by Using Multiple Scale Factors with Adaptive Thresholds.

    Science.gov (United States)

    Yang, Yingbao; Li, Xiaolong; Pan, Xin; Zhang, Yong; Cao, Chen

    2017-04-01

    Many downscaling algorithms have been proposed to address the issue of coarse-resolution land surface temperature (LST) derived from available satellite-borne sensors. However, few studies have focused on improving LST downscaling in urban areas with several mixed surface types. In this study, LST was downscaled by a multiple linear regression model between LST and multiple scale factors in mixed areas with three or four surface types. The correlation coefficients (CCs) between LST and the scale factors were used to assess the importance of the scale factors within a moving window. CC thresholds determined which factors participated in the fitting of the regression equation. The proposed downscaling approach, which involves an adaptive selection of the scale factors, was evaluated using the LST derived from four Landsat 8 thermal imageries of Nanjing City in different seasons. Results of the visual and quantitative analyses show that the proposed approach achieves relatively satisfactory downscaling results on 11 August, with coefficient of determination and root-mean-square error of 0.87 and 1.13 °C, respectively. Relative to other approaches, our approach shows the similar accuracy and the availability in all seasons. The best (worst) availability occurred in the region of vegetation (water). Thus, the approach is an efficient and reliable LST downscaling method. Future tasks include reliable LST downscaling in challenging regions and the application of our model in middle and low spatial resolutions.

  15. Shielding factors for gamma radiation from activity deposited on structures and ground surfaces

    International Nuclear Information System (INIS)

    Hedemann Jensen, P.

    1982-11-01

    This report describes a computer model that calculates shielding factors for indoor residence in multistorey and single-family houses for gamma radiation from activity despoited on roofs, outer walls, and ground surfaces. The dimensions of the buildings including window areas and the nearby surroundings has to be speficied in the calculations. Shielding factors can be calculated for different photon energies and for a uniform surface activity distribution as well as for separate activity on roof, outer wall, and ground surface achieved from decontamination or different deposition velocities. For a given area with a known distribution of different houses a weighted shielding factor can be calculated as well as a time-averaged one based on a given residence time distribution for work/school, home, outdoors, and transportation. Calculated shielding factors are shown for typical Danish houses. To give an impression of the sensitivity of the shielding factor on the parameters used in the model, variations were made in some of the most important parameters: wall thickness, road and ground width, percentage of outer wall covered by windows, photon energy, and decontamination percentage for outer walls, ground and roofs. The uncertainity of the calculations is discussed. (author)

  16. Radiation immobilization of catalase and its application

    International Nuclear Information System (INIS)

    Wang Guanghui; Ha Hongfei; Wang Xia; Wu Jilan

    1988-01-01

    Catalase was immobilized by a chemical method on porous polyacrylamide particles produced by radiation polymerization of acrylamide monomer at low temperature (-78 0 C). Activity of immobilized catalase was enhanced distinctly by joining a chemical arm to the support. The method of recovery of catalase activity on immobilized polymer was found by soaking it in certain buffer. The treatment of H 2 O 2 both in aqueous solution and alcoholic solution by using the immobilized catalase was performed. (author)

  17. One-pot preparation of conducting composite containing abundant amino groups on electrode surface for electrochemical detection of von willebrand factor

    Science.gov (United States)

    Wang, Wen; Ma, Chao; Li, Yi; Liu, Baihui; Tan, Liang

    2018-03-01

    A one-pot protocol based on cyclic voltammetric scan was employed to prepare new conducting composite that was abundant in amino groups. The scanning electron microscope, atomic force microscope, X-ray photoelectron spectroscopy and infrared spectrum characterization demonstrate that poly(azure A), gold nanoparticles, chitosan and cysteine were immobilized simultaneously on glassy carbon electrode surface. Von Willebrand factor (vWF) antibody (Ab) was subsequently assembled by using glutaraldehyde to construct the Ab/composite-modified electrode. The capture of vWF could inhibit the charge transfer between the ferri-/ferrocyanide probe and the electrode and exert the negative effect on the electrochemical response of the dye polymer in the conducting composite due to the strong steric hindrance effect. The DPV peak current change before and after the immunoreaction was found to be proportional to the logarithm of the vWF concentration from 0.001 to 100 μg mL-1 with a detection limit of 0.4 ng mL-1. The proposed label-free electrochemical method was employed in the investigation on the release of vWF by oxidation-injured vascular endothelial cells. The experimental results exhibit that the vWF content in growth medium was increased when the oxidation injury of the cells was intensified in the presence of H2O2.

  18. Early halo immobilization of displaced traumatic spondylolisthesis of the axis.

    Science.gov (United States)

    Vaccaro, Alexander R; Madigan, Luke; Bauerle, Wayne B; Blescia, Adam; Cotler, Jerome M

    2002-10-15

    A retrospective study evaluating early halo immobilization of Types II and IIA hangman's fractures. To determine the treatment efficacy of early halo immobilization of Type II and IIA hangman's fractures. The treatment of hangman's fractures with traction reduction is well established, but the time required in traction before the institution of halo-vest immobilization is controversial. A retrospective review of all patients admitted to a level one spinal cord injury center between 1986 and 1999 with either a Type II or IIA hangman's fracture was performed. Initial and final radiographs were measured for translation and angulation. The need for reapplication of traction was also recorded. There were a total of 27 Type II and four Type IIA fractures. Of the Type II fractures, 21 went onto union after early halo immobilization. Six patients required reapplication of traction reduction because of fracture displacement. The Type IIA fractures all progressed to union. The discerning factor between the failure group and the success group was the initial degree of angulation on lateral cervical radiography. The patients requiring reapplication of traction had an initial fracture angulation of 12 degrees or greater. Early halo immobilization after traction reduction of Type II and IIA hangman's fractures is an effective method of management. Type II fractures with an angulation of greater than or equal to 12 degrees may require an extended period of traction to ensure adequate long-term fracture alignment.

  19. A review and assessment of the stress-intensity factors for surface cracks

    Science.gov (United States)

    Newman, J. C., Jr.

    1979-01-01

    The stress-intensity factor solutions proposed for a surface crack in a finite plate subjected to uniform tension are reviewed. Fourteen different solutions obtained over the past 16 years using approximate analytical methods, experimental methods, and engineering estimates are compared. The accuracy of the various solutions is assessed by correlating fracture data on surface-cracked tension specimens made of a brittle epoxy material. Fracture of the epoxy material was characterized by a constant value of stress-intensity factor as failure. Thus, the correctness of various solutions is judged by the variations in the stress-intensity factors at failure. The solutions were ranked in order of minimum standard deviation. The highest ranking solutions correlated 95% of data analyzed within + or - 10% whereas the lowest ranking solutions correlated 95% of data analyzed within + or - 20%. Some solutions could be applied to all data considered, whereas others were limited with respect to crack shapes and crack sizes that could be analyzed.

  20. Modification of transition's factor in the compact surface-potential-based MOSFET model

    Directory of Open Access Journals (Sweden)

    Kevkić Tijana

    2016-01-01

    Full Text Available The modification of an important transition's factor which enables continual behavior of the surface potential in entire useful range of MOSFET operation is presented. The various modifications have been made in order to obtain an accurate and computationally efficient compact MOSFET model. The best results have been achieved by introducing the generalized logistic function (GL in fitting of considered factor. The smoothness and speed of the transition of the surface potential from the depletion to the strong inversion region can be controlled in this way. The results of the explicit model with this GL functional form for transition's factor have been verified extensively with the numerical data. A great agreement was found for a wide range of substrate doping and oxide thickness. Moreover, the proposed approach can be also applied on the case where quantum mechanical effects play important role in inversion mode.

  1. Laser-assisted immobilization of colloid silver nanoparticles on polyethyleneterephthalate

    Czech Academy of Sciences Publication Activity Database

    Siegel, J.; Lyutakov, O.; Polívková, M.; Staszek, M.; Hubáček, Tomáš; Švorčík, V.

    2017-01-01

    Roč. 420, OCT (2017), s. 661-668 ISSN 0169-4332 R&D Projects: GA MŠk LM2015075 Institutional support: RVO:60077344 Keywords : silver nanoparticles * polyethyleneterephthalate * excimer laser * immobilization Subject RIV: JJ - Other Materials OBOR OECD: Materials engineering Impact factor: 3.387, year: 2016

  2. Immobilization of technetium and nitrate in cement-based materials

    International Nuclear Information System (INIS)

    Tallent, O.K.; McDaniel, E.W.; Del Cul, G.D.; Dodson, K.E.; Trotter, D.R.

    1987-01-01

    The leachabilities of technetium and nitrate wastes immobilized in cement-based grouts have been investigated. Factors found to affect the leachabilities include grout mix ratio, grout fluid density, dry solid blend composition, and waste concentration. 10 refs., 7 figs., 3 tabs

  3. Method of immobilizing carbon dioxide from gas streams

    International Nuclear Information System (INIS)

    Holladay, D.W.; Haag, G.L.

    1979-01-01

    This invention comprises a method for rapidly and continuously immobilizing carbon dioxide contained in various industrial off-gas streams, the carbon dioxide being immobilized as dry, stable, and substantially water-insoluble particulates. Briefly, the method comprises passing the gas stream through a fixed or fluidized bed of hydrated barium hydroxide to remove and immobilize the carbon dioxide by converting the bed to barium carbonate. The method has several important advantages: it can be conducted effectively at ambient temperature; it provides a very rapid reaction rate over a wide range of carbon dioxide concentrations; it provides high decontamination factors; and it has a high capacity for carbon dioxide. The invention is especially well suited for the removal of radioactive carbon dioxide from off-gases generated by nuclear-fuel reprocessing facilities and nuclear power plants

  4. Immobilization and characterization of inulinase from Ulocladium

    Indian Academy of Sciences (India)

    Ulocladium atrum inulinase was immobilized on different composite membranes composed of chitosan/nonwoven fabrics. Km values of free and immobilized U. atrum inulinase on different composite membranes were calculated. The enzyme had optimum pH at 5.6 for free and immobilized U. atrum inulinase on polyester ...

  5. Improved methane removal in exhaust gas from biogas upgrading process using immobilized methane-oxidizing bacteria.

    Science.gov (United States)

    Sun, Meng-Ting; Yang, Zhi-Man; Fu, Shan-Fei; Fan, Xiao-Lei; Guo, Rong-Bo

    2018-05-01

    Methane in exhaust gas from biogas upgrading process, which is a greenhouse gas, could cause global warming. The biofilter with immobilized methane-oxidizing bacteria (MOB) is a promising approach for methane removal, and the selections of inoculated MOB culture and support material are vital for the biofilter. In this work, five MOB consortia were enriched at different methane concentrations. The MOB-20 consortium enriched at the methane concentration of 20.0% (v/v) was then immobilized on sponge and two particle sizes of volcanic rock in biofilters to remove methane in exhaust gas from biogas upgrading process. Results showed that the immobilized MOB performed more admirable methane removal capacity than suspended cells. The immobilized MOB on sponge reached the highest methane removal efficiency (RE) of 35%. The rough surface, preferable hydroscopicity, appropriate pore size and particle size of support material might favor the MOB immobilization and accordingly methane removal. Copyright © 2018 Elsevier Ltd. All rights reserved.

  6. A Versatile Microarray Immobilization Strategy Based on a Biorthogonal Reaction Between Tetrazine and Trans-Cyclooctene.

    Science.gov (United States)

    Wang, Ping; Gao, Liqian; Lei, Haipeng; Lee, Su Seong; Yao, Shao Q; Sun, Hongyan

    2017-01-01

    Given its increasing importance in transforming biomedical research in recent years, microarray technology has become highly popular as a powerful screening platform in detecting biomolecule interactions, discovering new inhibitors, and identifying biomarkers as well as diagnosing disease. The success of microarray technology in various biological applications is highly dependent on the accessibility, the functionality, and the density of the surface bound biomolecules. Therefore, compound immobilization represents a critical step for the successful implementation of microarray screening. Herein we describe a fast and site-specific microarray immobilization approach by using trans-cyclooctene-tetrazine ligation. This approach not only ensures fast immobilization and uniform display of biomolecules, but also allows the optimum orientation of biomolecules after immobilization. All these excellent properties facilitate subsequent interactions of the biomolecules and their interacting partners during the screening process. We envision that the immobilization strategy described here can find useful applications in many other microarray related studies.

  7. Evaluation of immobilized-lysozyme by means of TOF-SIMS

    Science.gov (United States)

    Okada, Keigo; Aoyagi, Satoka; Dohi, Makoto; Kato, Nobuhiko; Kudo, Masahiro; Tozu, Miyako; Miyayama, Takuya; Sanada, Noriaki

    2008-12-01

    Evaluation of immobilized-proteins on bio-devices is important for the development of sophisticated devices. Lysozyme molecules immobilized on substrates were evaluated by means of time-of-flight secondary ion mass spectrometry (TOF-SIMS). Two types of the lysozyme-immobilized samples were prepared by controlling the binding parts, i.e., the amino groups or carboxyl groups, of the protein. The TOF-SIMS spectra of each sample were analyzed with mutual information to select fragment ions specific to each sample. According to the results, differences between the samples being immobilized in the different ways are suggested, and the surface structure of the lysozyme molecule immobilized at amino groups is determined based on three-dimensional structure of lysozyme in the Protein Data Bank.

  8. Frozen Microemulsions for MAPLE Immobilization of Lipase

    Directory of Open Access Journals (Sweden)

    Valeria Califano

    2017-12-01

    Full Text Available Candida rugosa lipase (CRL was deposited by matrix assisted pulsed laser evaporation (MAPLE in order to immobilize the enzyme with a preserved native conformation, which ensures its catalytic functionality. For this purpose, the composition of the MAPLE target was optimized by adding the oil phase pentane to a water solution of the amino acid 3-(3,4-dihydroxyphenyl-2-methyl-l-alanine (m-DOPA, giving a target formed by a frozen water-lipase-pentane microemulsion. Fourier transform infrared (FTIR spectroscopy and atomic force microscopy (AFM were used to investigate the structure of MAPLE deposited lipase films. FTIR deconvolution of amide I band indicated a reduction of unfolding and aggregation, i.e., a better preserved lipase secondary structure in the sample deposited from the frozen microemulsion target. AFM images highlighted the absence of big aggregates on the surface of the sample. The functionality of the immobilized enzyme to promote transesterification was determined by thin layer chromatography, resulting in a modified specificity.

  9. Surface Irregularity Factor as a Parameter to Evaluate the Fatigue Damage State of CFRP

    Directory of Open Access Journals (Sweden)

    Pablo Zuluaga-Ramírez

    2015-11-01

    Full Text Available This work presents an optical non-contact technique to evaluate the fatigue damage state of CFRP structures measuring the irregularity factor of the surface. This factor includes information about surface topology and can be measured easily on field, by techniques such as optical perfilometers. The surface irregularity factor has been correlated with stiffness degradation, which is a well-accepted parameter for the evaluation of the fatigue damage state of composite materials. Constant amplitude fatigue loads (CAL and realistic variable amplitude loads (VAL, representative of real in- flight conditions, have been applied to “dog bone” shaped tensile specimens. It has been shown that the measurement of the surface irregularity parameters can be applied to evaluate the damage state of a structure, and that it is independent of the type of fatigue load that has caused the damage. As a result, this measurement technique is applicable for a wide range of inspections of composite material structures, from pressurized tanks with constant amplitude loads, to variable amplitude loaded aeronautical structures such as wings and empennages, up to automotive and other industrial applications.

  10. Dose behind various immobilization and beam-modifying devices

    International Nuclear Information System (INIS)

    Mellenberg, David E.

    1995-01-01

    Purpose: To quantify the degradation of skin sparing associated with using beam modifiers such as compensators, immobilization devices, and custom blocks for high energy photon beams. Methods and Materials: The degradation of skin sparing was quantified by measuring dose build-up curves with an extrapolation chamber for 6 and 15 MV photon beams. Uniform thickness compensators made of gypsum and lead, thermoplastic mask material, immobilization cradle foam, and cerrobend custom blocks were placed in geometries that mimic relevant clinical situations. Results: Compensators, whether made of gypsum or lead, placed in the linear accelerator's wedge slot did not significantly effect the depth dose curve's build-up region. Immobilization devices such as cradle foam or thermoplastic placed in contact with the patient degrade the skin sparing expected from high energy photon beams proportional to their thickness and density. Measurements behind custom blocks show that surface and near surface doses for a blocked field are best described by build-up curves for an equivalent size open field. Conclusions: These results allow explanation and possibly prediction of skin reactions on patients in which compensators, foam immobilization cradles, thermoplastic masks, or custom blocks are used. These results also provide a baseline by which substitute materials can be evaluated

  11. SHINE transcription factors act redundantly to pattern the archetypal surface of Arabidopsis flower organs.

    Directory of Open Access Journals (Sweden)

    Jian Xin Shi

    2011-05-01

    Full Text Available Floral organs display tremendous variation in their exterior that is essential for organogenesis and the interaction with the environment. This diversity in surface characteristics is largely dependent on the composition and structure of their coating cuticular layer. To date, mechanisms of flower organ initiation and identity have been studied extensively, while little is known regarding the regulation of flower organs surface formation, cuticle composition, and its developmental significance. Using a synthetic microRNA approach to simultaneously silence the three SHINE (SHN clade members, we revealed that these transcription factors act redundantly to shape the surface and morphology of Arabidopsis flowers. It appears that SHNs regulate floral organs' epidermal cell elongation and decoration with nanoridges, particularly in petals. Reduced activity of SHN transcription factors results in floral organs' fusion and earlier abscission that is accompanied by a decrease in cutin load and modified cell wall properties. SHN transcription factors possess target genes within four cutin- and suberin-associated protein families including, CYP86A cytochrome P450s, fatty acyl-CoA reductases, GSDL-motif lipases, and BODYGUARD1-like proteins. The results suggest that alongside controlling cuticular lipids metabolism, SHNs act to modify the epidermis cell wall through altering pectin metabolism and structural proteins. We also provide evidence that surface formation in petals and other floral organs during their growth and elongation or in abscission and dehiscence through SHNs is partially mediated by gibberellin and the DELLA signaling cascade. This study therefore demonstrates the need for a defined composition and structure of the cuticle and cell wall in order to form the archetypal features of floral organs surfaces and control their cell-to-cell separation processes. Furthermore, it will promote future investigation into the relation between the

  12. SHINE transcription factors act redundantly to pattern the archetypal surface of Arabidopsis flower organs.

    Science.gov (United States)

    Shi, Jian Xin; Malitsky, Sergey; De Oliveira, Sheron; Branigan, Caroline; Franke, Rochus B; Schreiber, Lukas; Aharoni, Asaph

    2011-05-01

    Floral organs display tremendous variation in their exterior that is essential for organogenesis and the interaction with the environment. This diversity in surface characteristics is largely dependent on the composition and structure of their coating cuticular layer. To date, mechanisms of flower organ initiation and identity have been studied extensively, while little is known regarding the regulation of flower organs surface formation, cuticle composition, and its developmental significance. Using a synthetic microRNA approach to simultaneously silence the three SHINE (SHN) clade members, we revealed that these transcription factors act redundantly to shape the surface and morphology of Arabidopsis flowers. It appears that SHNs regulate floral organs' epidermal cell elongation and decoration with nanoridges, particularly in petals. Reduced activity of SHN transcription factors results in floral organs' fusion and earlier abscission that is accompanied by a decrease in cutin load and modified cell wall properties. SHN transcription factors possess target genes within four cutin- and suberin-associated protein families including, CYP86A cytochrome P450s, fatty acyl-CoA reductases, GSDL-motif lipases, and BODYGUARD1-like proteins. The results suggest that alongside controlling cuticular lipids metabolism, SHNs act to modify the epidermis cell wall through altering pectin metabolism and structural proteins. We also provide evidence that surface formation in petals and other floral organs during their growth and elongation or in abscission and dehiscence through SHNs is partially mediated by gibberellin and the DELLA signaling cascade. This study therefore demonstrates the need for a defined composition and structure of the cuticle and cell wall in order to form the archetypal features of floral organs surfaces and control their cell-to-cell separation processes. Furthermore, it will promote future investigation into the relation between the regulation of organ

  13. Functionalization of silicone rubber for the covalent immobilization of fibronectin.

    Science.gov (United States)

    Völcker, N; Klee, D; Höcker, H; Langefeld, S

    2001-02-01

    Surface modification techniques were employed in order to provide functionalized silicone rubber with enhanced cytocompatibility. Acrylic acid (AAc), methacrylic acid (MAAc) and glycidylmethacrylate (GMA) were graft-co-polymerized onto the surface of silicone induced by an argon plasma and thermal initiation. The polymerizations were carried out in solution, in the case of acrylic acid a vapor phase graft-co-polymerization subsequent to argon plasma activation was carried out as well. Human fibronectin (hFn), which acts as a cell adhesion mediator for fibroblasts, was immobilized by making use of the generated carboxylic or epoxy groups, respectively. Surface analysis was accomplished by means of X-ray photoelectron spectroscopy (XPS), infrared spectroscopy in attenuated total reflection mode (IR-ATR), scanning electron microscopy (SEM), atomic force microscopy (AFM) and dynamic contact angle measurements using the Wilhelmy-plate method. The amount of immobilized active hFn was semiquantified by enzyme-linked immunosorbent assay (ELISA) using a structure-specific antibody against the cell-binding domain of hFn. In vitro testing showed a remarkable difference between surfaces exposing adsorbed-only and surfaces with covalently immobilized hFn. Copyright 2001 Kluwer Academic Publishers

  14. The mechanisms of heavy metal immobilization by cementitious material treatments and thermal treatments: A review.

    Science.gov (United States)

    Guo, Bin; Liu, Bo; Yang, Jian; Zhang, Shengen

    2017-05-15

    Safe disposal of solid wastes containing heavy metals is a significant task for environment protection. Immobilization treatment is an effective technology to achieve this task. Cementitious material treatments and thermal treatments are two types of attractive immobilization treatments due to that the heavy metals could be encapsulated in their dense and durable wasteforms. This paper discusses the heavy metal immobilization mechanisms of these methods in detail. Physical encapsulation and chemical stabilization are two fundamental mechanisms that occur simultaneously during the immobilization processes. After immobilization treatments, the wasteforms build up a low permeable barrier for the contaminations. This reduces the exposed surface of wastes. Chemical stabilization occurs when the heavy metals transform into more stable and less soluble metal bearing phases. The heavy metal bearing phases in the wasteforms are also reviewed in this paper. If the heavy metals are incorporated into more stable and less soluble metal bearing phases, the potential hazards of heavy metals will be lower. Thus, converting heavy metals into more stable phases during immobilization processes should be a common way to enhance the immobilization effect of these immobilization methods. Copyright © 2017 Elsevier Ltd. All rights reserved.

  15. Graphene immobilized enzyme/polyethersulfone mixed matrix membrane: Enhanced antibacterial, permeable and mechanical properties

    Science.gov (United States)

    Duan, Linlin; Wang, Yuanming; Zhang, Yatao; Liu, Jindun

    2015-11-01

    Enzyme immobilization has been developed to address lots of issues of free enzyme, such as instability, low activity and difficult to retain. In this study, graphene was used as an ideal carrier for lysozyme immobilization, including graphene oxide (GO) immobilized lysozyme (GO-Ly) and chemically reduced graphene oxide (CRGO) immobilized lysozyme (CRGO-Ly). Herein, lysozyme as a bio-antibacterial agent has excellent antibacterial performance and the products of its catalysis are safety and nontoxic. Then the immobilized lysozyme materials were blended into polyethersulfone (PES) casting solution to prepare PES ultrafiltration membrane via phase inversion method. GO and CRGO were characterized by Fourier transform infrared spectroscopy (FTIR), Ultraviolet-visible spectrum (UV), X-ray diffraction (XRD), and transmission electron microscopy (TEM) and the immobilized lysozyme composites were observed by fluorescent microscopy. The results revealed that GO and CRGO were successfully synthesized and lysozyme was immobilized on their surfaces. The morphology, hydrophilicity, mechanical properties, separation properties and antibacterial activity of the hybrid membranes were characterized in detail. The hydrophilicity, water flux and mechanical strength of the hybrid membranes were significantly enhanced after adding the immobilized lysozyme. In the antibacterial experiment, the hybrid membranes exhibited an effective antibacterial performance against Escherichia coli (E. coli).

  16. Cadmium immobilization by hydroxyapatite

    Directory of Open Access Journals (Sweden)

    Smičiklas Ivana D.

    2003-01-01

    Full Text Available The contamination of air, soil and water by cadmium is a great environmental problem. If cadmium occurs in nature in ionic form, soluble in water, it easily enters into the food chain. Hydroxyapatite (HAP, Ca-o(POAe(OH2 is a sparingly soluble salt and an excellent matrix for the removal of heavy metals from solutions. Considerable research attention has been paid to the bond between Cc/2+ ions and synthetic hydroxyapatite of known composition. The sorption mechanism is complex. The dominant process is ion exchange, but surface adsorption, surface complexation and coprecipitation can also contribute to the overall mechanism. The sorption capacity depends on the characteristics of hydroxyapatite itself and on the experimental conditions. Under optimum conditions a maximum capacity of 0.8 mol Cd2+/mol HAP can be achieved. HAP is a potential sorbent for the remediation of contaminated water and soil, for industrial waste treatment, and it is also referenced as a material that can be used as a barrier around waste depositories.

  17. Covalent immobilization of xylanase produced from Bacillus pumilus SV-85S on electrospun polymethyl methacrylate nanofiber membrane.

    Science.gov (United States)

    Kumar, Pankaj; Gupta, Ashish; Dhakate, Sanjay R; Mathur, Rakesh B; Nagar, Sushil; Gupta, Vijay K

    2013-01-01

    Polymethyl methacrylate (PMMA) nanofiber membrane (NFM) was synthesized by an electrospinning technique. These membranes were utilized as a support for immobilization of xylanase enzyme to study its pH stability, thermal stability, and reusability. The morphology of aligned NFM was studied by optical microscopy and scanning electron microscopy. The PMMA NFM was functionalized with phenylenediamine and activated with glutaraldehyde to yield an aldehyde group on its surface for covalent immobilization of xylanase. The Fourier transform infrared analysis of the covalently immobilized xylanase confirmed that the enzyme was immobilized on PMMA NFM via amide linkages. The immobilization efficiency of covalently bound xylanase was found experimentally to be 90%. A forward shift in pH optima from 6.0-7.0 (soluble enzyme) to 7.0-9.0 (immobilized enzyme) was observed after xylanase immobilization. The pH and temperature stability of xylanase were enhanced upon its covalent immobilization. The immobilized enzyme was active on repeated use and retained ∼80% of its initial activity after 11 reaction cycles. The improved thermal and operational stability of the covalently immobilized enzyme on PMMA NFM might be advantageous for industrial applications. © 2013 International Union of Biochemistry and Molecular Biology, Inc.

  18. A facile and efficient method of enzyme immobilization on silica particles via Michael acceptor film coatings: immobilized catalase in a plug flow reactor.

    Science.gov (United States)

    Bayramoglu, Gulay; Arica, M Yakup; Genc, Aysenur; Ozalp, V Cengiz; Ince, Ahmet; Bicak, Niyazi

    2016-06-01

    A novel method was developed for facile immobilization of enzymes on silica surfaces. Herein, we describe a single-step strategy for generating of reactive double bonds capable of Michael addition on the surfaces of silica particles. This method was based on reactive thin film generation on the surfaces by heating of impregnated self-curable polymer, alpha-morpholine substituted poly(vinyl methyl ketone) p(VMK). The generated double bonds were demonstrated to be an efficient way for rapid incorporation of enzymes via Michael addition. Catalase was used as model enzyme in order to test the effect of immobilization methodology by the reactive film surface through Michael addition reaction. Finally, a plug flow type immobilized enzyme reactor was employed to estimate decomposition rate of hydrogen peroxide. The highly stable enzyme reactor could operate continuously for 120 h at 30 °C with only a loss of about 36 % of its initial activity.

  19. Metal organic frameworks for enzyme immobilization in biofuel cells

    Science.gov (United States)

    Bodell, JaDee

    Interest in biofuel cells has been rapidly expanding as an ever-growing segment of the population gains access to electronic devices. The largest areas of growth for new populations using electronic devices are often in communities without electrical infrastructure. This lack of infrastructure in remote environments is one of the key driving factors behind the development of biofuel cells. Biofuel cells employ biological catalysts such as enzymes to catalyze oxidation and reduction reactions of select fuels to generate power. There are several benefits to using enzymes to catalyze reactions as compared to traditional fuel cells which use metal catalysts. First, enzymes are able to catalyze reactions at or near room temperature, whereas traditional metal catalysts are only efficient at very high temperatures. Second, biofuel cells can operate under mild pH conditions which is important for the eventual design of safe, commercially viable devices. Also, biofuel cells allow for implantable and flexible technologies. Finally, enzymes exhibit high selectivity and can be combined to fully oxidize or reduce the fuel which can generate several electrons from a single molecule of fuel, increasing the overall device efficiency. One of the main challenges which persist in biofuel cells is the instability of enzymes over time which tend to denature after hours or days. For a viable commercial biofuel cell to be produced, the stability of enzymes must be extended to months or years. Enzymes have been shown to have improved stability after being immobilized. The focus of this research was to find a metal organic framework (MOF) structure which could successfully immobilize enzymes while still allowing for electron transport to occur between the catalytic center of the enzyme and the electrode surface within a biofuel cell for power generation. Four MOF structures were successfully synthesized and were subsequently tested to determine the MOF's ability to immobilize the following

  20. Root caries, root surface restorations and lifestyle factors in adult Danes

    DEFF Research Database (Denmark)

    Christensen, Lisa Bøge; Jensen, Allan Bardow; Ekstrand, Kim

    2015-01-01

    AIM: To investigate selected lifestyle factors in relation to active caries and restored root surface lesions in adults. MATERIALS AND METHODS: Based on clinical examinations and questionnaires, data on root caries, socioeconomic status, body mass index, dietary habits, alcohol consumption, tobacco...... use and oral hygiene routines were collected from 4369 adults aged 21-89 who took part in a survey covering 13 municipalities across Denmark. Uni- and multivariate logistic regression analyses were applied to analyse the relationship between the independent lifestyle variables and active caries...... and restored root surface lesions, respectively. RESULTS: The prevalence of active root caries was 4%, while 26% displayed restored root surfaces. The sugar intake was not related to root caries. A multivariate logistic regression analysis revealed that, in subjects aged 45 or over, smoking and wearing...

  1. Biodiesel production with immobilized lipase: A review.

    Science.gov (United States)

    Tan, Tianwei; Lu, Jike; Nie, Kaili; Deng, Li; Wang, Fang

    2010-01-01

    Fatty acid alkyl esters, also called biodiesel, are environmentally friendly and show great potential as an alternative liquid fuel. Biodiesel is produced by transesterification of oils or fats with chemical catalysts or lipase. Immobilized lipase as the biocatalyst draws high attention because that process is "greener". This article reviews the current status of biodiesel production with immobilized lipase, including various lipases, immobilization methods, various feedstocks, lipase inactivation caused by short chain alcohols and large scale industrialization. Adsorption is still the most widely employed method for lipase immobilization. There are two kinds of lipase used most frequently especially for large scale industrialization. One is Candida antartica lipase immobilized on acrylic resin, and the other is Candida sp. 99-125 lipase immobilized on inexpensive textile membranes. However, to further reduce the cost of biodiesel production, new immobilization techniques with higher activity and stability still need to be explored. Copyright 2010 Elsevier Inc. All rights reserved.

  2. Determination of conversion factors and efficiency for GM detectors used in measurements of surface pollution

    International Nuclear Information System (INIS)

    Ayala G, J.; Alvarez R, J.T.

    1999-01-01

    One of the objectives of the Radiation protection, is to observe with the National and as International standardization referring to the ICRP dose limitation system (ICRP 26, ICRP 60). In this work it was treated the problem corresponding about how to determine the conversion factor of cpm/mR/h and the absolute efficiency ε, for a Geiger-Muller equipment with thin window. This equipment is used for the beta particle detection. Thus the correct use of calibration factors and the procedures to convert cpm in Bq is expedited and also to apply the ISO procedure for to evaluate contaminated surfaces. (Author)

  3. Tailoring Surface Impurity Content to Maximize Q-factors of Superconducting Resonators

    Energy Technology Data Exchange (ETDEWEB)

    Martinello, Martina [Illinois Inst. of Technology, Chicago, IL (United States); Fermi National Accelerator Lab. (FNAL), Batavia, IL (United States); Checchin, Mattia [Illinois Inst. of Technology, Chicago, IL (United States); Fermi National Accelerator Lab. (FNAL), Batavia, IL (United States); Grassellino, Anna [Fermi National Accelerator Lab. (FNAL), Batavia, IL (United States); Melnychuk, Oleksandr [Fermi National Accelerator Lab. (FNAL), Batavia, IL (United States); Posen, Sam [Fermi National Accelerator Lab. (FNAL), Batavia, IL (United States); Romanenko, Alexander [Fermi National Accelerator Lab. (FNAL), Batavia, IL (United States); Sergatskov, Dmitri [Fermi National Accelerator Lab. (FNAL), Batavia, IL (United States); Zasadzinski, John [Illinois Inst. of Technology, Chicago, IL (United States)

    2016-06-01

    Quality factor of superconducting radio-frequency (SRF) cavities is degraded whenever magnetic flux is trapped in the cavity walls during the cooldown. In this contribution we study how the trapped flux sensitivity, defined as the trapped flux surface resistance normalized for the amount of flux trapped, depends on the mean free path. A variety of 1.3 GHz cavities with different surface treatments (EP, 120 C bake and different N-doping) were studied in order to cover the largest range of mean free path nowadays achievable, from few to thousands of nanometers. A bell shaped trend appears for the range of mean free path studied. Over doped cavities falls at the maximum of this curve defining the largest values of sensitivity. In addition, we have also studied the trend of the BCS surface resistance contribution as a function of mean free path, revealing that N-doped cavities follow close to the theoretical minimum of the BCS surface resistance as a function of the mean free path. Adding these results together we unveil that optimal N-doping treatment allows to maximize Q-factor at 2 K and 16 MV/m until the magnetic field fully trapped during the cavity cooldown stays below 10 mG.

  4. Statistical Modelling and Optimization of Fermentation Medium for Lincomycin Production by Streptomyces lincolnensis Immobilized Cells

    Directory of Open Access Journals (Sweden)

    Nayera A.M. Abdelwahed

    2017-05-01

    Full Text Available ABSTRACT Response surface methodology was used to optimize lincomycin production by Streptomyces lincolnensis NRRL ISP-5355 in submerged fermentation. Screening of fermentation medium components to find their relative effect on lincomycin production was done using Plackett-Burman design. Malt extract, dextrin, soluble starch and (NH42SO4 were the most significant nutrient influenced on lincomycin production. Central composite design was applied to determine optimal concentrations of these factors and the effect of their mutual interactions. The interaction between soluble starch and (NH42SO4 was found to enhance the production, whereas malt extract and dextrin exhibited an influence independent from the other two factors. Using this statistical optimization method, maximum lincomycin concentration of 1345 μg/ml was obtained which represented a 40.5 % increase in titer than that acquired from the non-optimized medium. This statistically optimized medium was employed for lincomycin production through immobilization of Streptomyces lincolnensis by adsorption on synthetic cotton fibers. Immobilization technique improved the concentration to 1350 μg/ml higher than that produced from free cells cultures and could be maintained for longer than 17 days in a repeated batch system.

  5. Immobilization of olfactory receptors onto gold electrodes for electrical biosensor

    Energy Technology Data Exchange (ETDEWEB)

    Casuso, Ignacio [Departament d' Electronica, Universitat de Barcelona, Laboratori de Nanobioenginyeria-IBEC, Parc Cientific de Barcelona, C/Josep Samitier 1-5, Barcelona (Spain)], E-mail: icasuso@pcb.ub.es; Pla-Roca, Mateu [Departament d' Electronica, Universitat de Barcelona, Laboratori de Nanobioenginyeria-IBEC, Parc Cientific de Barcelona, C/Josep Samitier 1-5, Barcelona (Spain); Gomila, Gabriel [Departament d' Electronica, Universitat de Barcelona, Laboratori de Nanobioenginyeria-IBEC, Parc Cientific de Barcelona, C/Josep Samitier 1-5, Barcelona (Spain)], E-mail: ggomila@pcb.ub.es; Samitier, Josep [Departament d' Electronica, Universitat de Barcelona, Laboratori de Nanobioenginyeria-IBEC, Parc Cientific de Barcelona, C/Josep Samitier 1-5, Barcelona (Spain); Minic, Jasmina; Persuy, Marie A.; Salesse, Roland; Pajot-Augy, Edith [INRA, Neurobiologie de l' Olfaction et de la Prise Alimentaire, Equipe Recepteurs et Communication Chimique, Domaine de Vilvert, Jouy en Josas Cedex (France)

    2008-07-01

    We investigate the immobilization of native nanovesicles containing functional olfactory receptors onto gold electrodes by means of atomic force microscopy in liquid. We show that nanovesicles can be adsorbed without disrupting them presenting sizes once immobilized ranging from 50 nm to 200 nm in diameter. The size of the nanovesicles shows no dependence on the electrode hydrophobicity being constant in a height/width ratio close to 1:3. Nevertheless, electrode hydrophobicity does affect the surface coverage, the surface coverage is five times higher in hydrophilic electrodes than on hydrophobic ones. Surface coverage is also affected by nanovesicles dimensions in suspension, the size homogenization to around 50 nm yields a further five fold increment in surface coverage achieving a coverage of about 50% close to the hard spheres jamming limit (54.7%). A single layer of nanovesicles is always formed with no particle overlap. Present results provide insights into the immobilization on electrodes of olfactory receptors for further olfactory electrical biosensor development.

  6. Immobilization of chlorine dioxide modified cells for uranium absorption

    International Nuclear Information System (INIS)

    He, Shengbin; Ruan, Binbiao; Zheng, Yueping; Zhou, Xiaobin; Xu, Xiaoping

    2014-01-01

    There has been a trend towards the use of microorganisms to recover metals from industrial wastewater, for which various methods have been reported to be used to improve microorganism adsorption characteristics such as absorption capacity, tolerance and reusability. In present study, chlorine dioxide(ClO 2 ), a high-efficiency, low toxicity and environment-benign disinfectant, was first reported to be used for microorganism surface modification. The chlorine dioxide modified cells demonstrated a 10.1% higher uranium adsorption capacity than control ones. FTIR analysis indicated that several cell surface groups are involved in the uranium adsorption and cell surface modification. The modified cells were further immobilized on a carboxymethylcellulose (CMC) matrix to improve their reusability. The cell-immobilized adsorbent could be employed either in a high concentration system to move vast UO 2 2+ ions or in a low concentration system to purify UO 2 2+ contaminated water thoroughly, and could be repeatedly used in multiple adsorption-desorption cycles with about 90% adsorption capacity maintained after seven cycles. - Highlights: • Chlorine dioxide was first reported to be used for microorganism surface modification. • The chlorine dioxide modified cells demonstrated a 10.1% higher uranium adsorption capacity than control ones. • The chlorine dioxide modified cells were further immobilized by carboxymethylcellulose to improve their reusability

  7. The development, characterization, and application of biomimetic nanoscale enzyme immobilization

    Science.gov (United States)

    Haase, Nicholas R.

    The utilization of enzymes is of interest for applications such as biosensors and biofuel cells. Immobilizing enzymes provides a means to develop these applications. Previous immobilization efforts have been accomplished by exposing surfaces on which silica-forming molecules are present to solutions containing an enzyme and a silica precursor. This approach leads to the enzyme being entrapped in a matrix three orders of magnitude larger than the enzyme itself, resulting in low retention of enzyme activity. The research herein introduces a method for the immobilization of enzymes during the layer-by-layer buildup of Si-O and Ti-O coatings which are nanoscale in thickness. This approach is an application of a peptide-induced mineral deposition method developed in the Sandhage and Kroger groups, and it involves the alternating exposure of a surface to solutions containing the peptide protamine and then an aqueous precursor solution of silicon- or titanium-oxide at near-neutral pH. A method has been developed that enables in situ immobilization of enzymes in the protamine/mineral oxide coatings. Depending on the layer and mineral (silica or titania) within which the enzyme is incorporated, the resulting multilayer biocatalytic hybrid materials retain 20 -- 100% of the enzyme activity. Analyses of kinetic properties of the immobilized enzyme, coupled with characterization of physical properties of the mineral-bearing layers (thickness, porosity, pore size distribution), indicates that the catalytic activities of the enzymes immobilized in the different layers are largely determined by substrate diffusion. The enzyme was also found to be substantially stabilized against heat-induced denaturation and largely protected from proteolytic attack. These functional coatings are then developed for use as antimicrobial materials. Glucose oxidase, which catalyzes production of the cytotoxic agent hydrogen peroxide, was immobilized with silver nanoparticles, can release

  8. Effect of extracytoplasmic function sigma factors on autoaggregation, hemagglutination, and cell surface properties of Porphyromonas gingivalis

    Science.gov (United States)

    Kokubu, Eitoyo; Okamoto-Shibayama, Kazuko; Ishihara, Kazuyuki

    2017-01-01

    Porphyromonas gingivalis is a bacterium frequently isolated from chronic periodontal lesions and is involved in the development of chronic periodontitis. To colonize the gingival crevice, P. gingivalis has to adapt to environmental stresses. Microbial gene expression is regulated by transcription factors such as those in two-component systems and extracytoplasmic function (ECF) sigma factors. ECF sigma factors are involved in the regulation of environmental stress response genes; however, the roles of individual ECF sigma factors are largely unknown. The purpose of this study was to investigate the functions, including autoaggregation, hemagglutination, gingipain activity, susceptibility to antimicrobial agents, and surface structure formation, of P. gingivalis ECF sigma factors encoded by SigP (PGN_0274), SigCH (PGN_0319), PGN_0450, PGN_0970, and SigH (PGN_1740). Various physiological aspects of the sigP mutant were affected; autoaggregation was significantly decreased at 60 min (p < 0.001), hemagglutination activity was markedly reduced, and enzymatic activities of Kgp and Rgps were significantly decreased (p < 0.001). The other mutants also showed approximately 50% reduction in Rgps activity. Kgp activity was significantly reduced in the sigH mutant (p < 0.001). No significant differences in susceptibilities to tetracycline and ofloxacin were observed in the mutants compared to those of the wild-type strain. However, the sigP mutant displayed an increased susceptibility to ampicillin, whereas the PGN_0450 and sigH mutants showed reduced susceptibility. Transmission electron microscopy images revealed increased levels of outer membrane vesicles formed at the cell surfaces of the sigP mutant. These results indicate that SigP is important for bacterial surface-associated activities, including gingipain activity, autoaggregation, hemagglutination, vesicle formation, and antimicrobial susceptibility. PMID:28931045

  9. Emission factors for wind erosion of exposed aggregates at surface coal mines

    Energy Technology Data Exchange (ETDEWEB)

    Cowherd, C.

    1982-06-01

    The information presented in this paper is directed to those interested in inventorying dust emissions generated by wind erosion of exposed aggregates at surface coal mines. A testing program is described which entailed the use of a portable wind tunnel and an isokinetic sampling system to measure windblown dust emissions from coal and overburden materials at three western mine sites. Test measurements consisted of particle mass emission rates and size distributions for various control wind speeds and times after the initiation of wind erosion. The results indicate that natural surface crusts are very effective in mitigating suspended dust emissions and that a given surface has a finite potential for wind erosion subsequent to mechanical disturbance. Test data are used to develop a predictive emission factor equation which relates emission rate (per unit surface area) to the frequency of disturbance and the erosion potential corresponding to the fastest mile of wind for the period between disturbances. This equation can be used directly for flat surfaces or it can be coupled with an analysis of wind flow patterns around elevated storage piles to develop dust emission estimates for overall pile erosion.

  10. Laser induced forward transfer technique for the immobilization of biomaterials in biosensors applications (Conference Presentation)

    Science.gov (United States)

    Papazoglou, Symeon; Chatzipetrou, Marianeza; Massaouti, Maria; Zergioti, Ioanna

    2017-02-01

    Laser Induced Forward Transfer (LIFT) is a direct write technique, able to create micropatterns of biomaterials on sensing devices. In this conference we will present a new approach using LIFT for the printing and direct immobilization of biomaterials on a great variety of surfaces, for bio-sensor applications. The basic requirement for the fabrication of a biosensor is to stabilize a biomaterial that brings the physicochemical changes in close proximity to a transducer. In this direction, several immobilization methods such as covalent binding and crosslinking have been implemented. The presence of the additional functionalization steps in the biosensors fabrication, is among the main disadvantages of chemical immobilization methods. Our approach employs the LIFT technique for the direct immobilization of biomaterials, either by physical adsorption or by covalent bonding of the biomaterials. The physical adsorption of the biomaterials, occurs on hydrophobic or super-hydrophobic surfaces, due to the transition of the wetting properties of the surfaces upon the impact of the biomaterials with high velocity. The unique characteristic of LIFT technique to create high speed liquid jets, leads to the penetration of the biomaterial in the micro/nano roughness of the surface, resulting in their direct immobilization, without the need of any chemical functionalization layers. Moreover, we will also present the direct immobilization of biomaterials on Screen Printed Electrodes, for enzymatic biosensors, with a limit of detection (LOD) for catechol at 150 nM, and protein biosensors, used for the detection of herbicides, with an LOD of 8-10 nM.

  11. Stabilization of dimeric β-glucosidase from Aspergillus niger via glutaraldehyde immobilization under different conditions.

    Science.gov (United States)

    Vazquez-Ortega, Perla Guadalupe; Alcaraz-Fructuoso, Maria Teresa; Rojas-Contreras, Juan A; López-Miranda, Javier; Fernandez-Lafuente, Roberto

    2018-03-01

    The dimeric enzyme β-glucosidase from Aspergillus niger has been immobilized on different amino-agarose beads at pH 5 and 7, exploiting the versatility of glutaraldehyde. The stability of the free enzyme depended on enzyme concentration. Immobilization via ion exchange improved enzyme stability/activity, depending on the immobilization pH. However, the enzyme was desorbed in 75 mM NaCl at pH 7 and some stability/enzyme concentration dependence still existed. of these biocatalysts with glutaraldehyde increased enzyme stability (e.g. at pH 5, after incubation under conditions where the enzyme just ionically exchanged was fully inactivated, the activity of the glutaraldehyde treated enzyme remained unaltered). Immobilization on glutaraldehyde pre-activated supports yielded a higher increase in enzyme activity, but the stabilization was lower. While when measuring the enzyme activity at pH 4 there were no changes after immobilization, all immobilized enzymes were more active than the free enzyme at pH 6 and 7 (2-3 times). The Ki/Km ratio did not significantly decrease in any immobilized biocatalysts, and in some cases it worsened in a significant way (by a 9 fold factor using preactivated supports). The new biocatalysts are significantly more stable and avoid enzyme subunit desorption, being the immobilization pH a key point in their design. Copyright © 2017 Elsevier Inc. All rights reserved.

  12. Nanostructured ZnO-based biosensor: DNA immobilization and hybridization

    Directory of Open Access Journals (Sweden)

    Ahmed Mishaal Mohammed

    2017-09-01

    Full Text Available An electrochemical DNA biosensor was successfully fabricated by using (3-aminopropyl triethoxysilane (APTES with zinc oxide (ZnO nanorods synthesized using microwave-assisted chemical bath deposition method on thermally oxidized SiO2 thin films. The structural quality and morphology of the ZnO nanorods were determined by employing scanning electron microscopy (SEM and X-ray diffraction (XRD, which show a hexagonal wurtzite structure with a preferred orientation along the (101 direction. The surface of the SiO2 thin films was chemically modified with ZnO. Label-free detection DNA immobilization and hybridization were performed using potassium hexacyanoferrate with cyclic voltammetry (CV measurements. The capacitance, permittivity, and conductivity profiles of the fabricated sensor clearly indicate DNA immobilization and hybridization. Results show that the capacitance values of bare, ZnO- modified surface immobilization, and target DNA hybridization were 46 × 10−12 F, 47 × 10−8 F, 27 μF, and 17 μF, respectively, at 1 Hz. The permittivity measurement increased from 3.94 × 103 to 251 × 103 and 165 × 103 at the frequency range of approximately 200 to 1 Hz for bare and DNA immobilization and hybridization, respectively. The measured conductivity values for the bare, ZnO, immobilized, and hybridization device were 2.4 × 10−9, 10 × 10−8, 1.6 × 10−7, and 1.3 × 10−7 S cm−1, respectively.

  13. Perirenal fat surface area as a risk factor for morbidity after elective colorectal surgery.

    Science.gov (United States)

    Jung, Minoa; Volonté, Francesco; Buchs, Nicolas C; Gayet-Ageron, Angèle; Pugin, François; Gervaz, Pascal; Ris, Frederic; Morel, Philippe

    2014-02-01

    Visceral obesity appears to be an emerging parameter affecting postoperative outcome after abdominal surgery. However, total visceral fat remains time consuming to calculate, and there is still a lack of data about its value as an independent risk factor in colorectal surgery. The aim of this study was to validate the simple measurement of perirenal fat surface as a surrogate of visceral obesity, and to test the value of perirenal fat surface as a risk factor for morbidity in colorectal surgery and to compare it with the predictive value of other obesity parameters such as BMI and waist-hip ratio. This is a prospective observational cohort study. The study was conducted at a tertiary university hospital. Two hundred twenty-four consecutive patients (130 male) undergoing elective colorectal surgery with a mean age of 65.2 years (SD, ±12.9) were identified. Elective colorectal resections were performed. We assessed complications as the primary outcome measure. Secondary outcome measures were the conversion rates, duration of operation, and length of hospital stay. Perirenal fat surface was validated as a surrogate of visceral fat and a strong correlation between the 2 was confirmed (Spearman correlation coefficient ρ = 0.96). The overall postoperative complication rate was 22.8% (51/224) with 14.7% moderate complications (grade I and II) and 7.6% severe complications (grade III-IV), with a mortality rate of 0.5%. Multivariate analysis confirmed perirenal fat surface as an independent risk factor for postoperative complications (OR, 3.87; 95% CI, 1.73-8.64; p = 0.001), whereas BMI and waist-hip ratio were not statistically associated with postoperative complications (OR, 1.16; 95% CI, 0.51-2.66; p = 0.72). This study was limited by its sample size. Perirenal fat surface is an excellent and easy-to-reproduce indicator of visceral fat volume. Furthermore, perirenal fat surface is an independent risk factor for postoperative outcome in colorectal surgery that appears

  14. Specific and reversible immobilization of histidine-tagged proteins on functionalized silicon nanowires

    DEFF Research Database (Denmark)

    Liu, Yi-Chi; Rieben, Nathalie Ines; Iversen, Lars

    2010-01-01

    recognition motif attached to the Si NWs. In this study, we show that SiNWs can be chemically functionalized with Ni:NTA motifs, suitable for the specific immobilization of proteins via a short polyhistidine tag (His-tag) at close proximity to the SiNW surface. We demonstrate that the proteins preserve...... their function upon immobilization onto SiNWs. Importantly, the protein immobilization on the Si NWs is shown to be reversible after addition of EDTA or imidazole, thus allowing the regeneration of the bioFET when needed, such as in the case of proteins having a limited lifetime. We anticipate that our...

  15. Immobilization of iodine in concrete

    Science.gov (United States)

    Clark, Walter E.; Thompson, Clarence T.

    1977-04-12

    A method for immobilizing fission product radioactive iodine recovered from irradiated nuclear fuel comprises combining material comprising water, Portland cement and about 3-20 wt. % iodine as Ba(IO.sub.3).sub.2 to provide a fluid mixture and allowing the fluid mixture to harden, said Ba(IO.sub.3).sub.2 comprising said radioactive iodine. An article for solid waste disposal comprises concrete prepared by this method. BACKGROUND OF THE INVENTION This invention was made in the course of, or under a contract with the Energy Research and Development Administration. It relates in general to reactor waste solidification and more specifically to the immobilization of fission product radioactive iodine recovered from irradiated nuclear fuel for underground storage.

  16. Nuclear waste immobilization. Progress report

    Energy Technology Data Exchange (ETDEWEB)

    Ringwood, A.E.; Sinclair, W.; McLaughlin, G.M.

    1979-11-20

    United States defense nuclear wastes are presently in tank storage, largely as sludges comprising Fe, Mn, Ni, U and Na oxides and hydroxides, together with 0.5 to 5 percent of fission products and actinides (exclusive of uranium). The relative proportions of Al, Fe, Mn, Ni, U and Na in the sludges from different tanks vary considerably, except that (Fe + Al + Mn) are by far the major components and Fe is more abundant than Mn. Typical compositions of some calcined sludges from Savannah River are given. This paper briefly describes how the SYNROC process, utilizing straightforward technology, can be readily adapted to the problem of defense waste immobilization, yielding a dense, inert, ceramic waste-form, SYNROC-D. Two classes of processes are discussed - one designed to immobilize sludges containing normal amounts of sodium and the other designed for otherwise similar sludges which are, however, strongly depleted in sodium as a result of more efficient washing procedures.

  17. The 1.7 Å X-ray crystal structure of the porcine factor VIII C2 domain and binding analysis to anti-human C2 domain antibodies and phospholipid surfaces.

    Directory of Open Access Journals (Sweden)

    Caileen M Brison

    Full Text Available The factor VIII C2 domain is essential for binding to activated platelet surfaces as well as the cofactor activity of factor VIII in blood coagulation. Inhibitory antibodies against the C2 domain commonly develop following factor VIII replacement therapy for hemophilia A patients, or they may spontaneously arise in cases of acquired hemophilia. Porcine factor VIII is an effective therapeutic for hemophilia patients with inhibitor due to its low cross-reactivity; however, the molecular basis for this behavior is poorly understood. In this study, the X-ray crystal structure of the porcine factor VIII C2 domain was determined, and superposition of the human and porcine C2 domains demonstrates that most surface-exposed differences cluster on the face harboring the "non-classical" antibody epitopes. Furthermore, antibody-binding results illustrate that the "classical" 3E6 antibody can bind both the human and porcine C2 domains, although the inhibitory titer to human factor VIII is 41 Bethesda Units (BU/mg IgG versus 0.8 BU/mg IgG to porcine factor VIII, while the non-classical G99 antibody does not bind to the porcine C2 domain nor inhibit porcine factor VIII activity. Further structural analysis of differences between the electrostatic surface potentials suggest that the C2 domain binds to the negatively charged phospholipid surfaces of activated platelets primarily through the 3E6 epitope region. In contrast, the G99 face, which contains residue 2227, should be distal to the membrane surface. Phospholipid binding assays indicate that both porcine and human factor VIII C2 domains bind with comparable affinities, and the human K2227A and K2227E mutants bind to phospholipid surfaces with similar affinities as well. Lastly, the G99 IgG bound to PS-immobilized factor VIII C2 domain with an apparent dissociation constant of 15.5 nM, whereas 3E6 antibody binding to PS-bound C2 domain was not observed.

  18. Immobilization of iodine in concrete

    International Nuclear Information System (INIS)

    Clark, W.E.; Thompson, C.T.

    1977-01-01

    A method for immobilizing fission product radioactive iodine recovered from irradiated nuclear fuel comprises combining material comprising water, Portland cement and about 3 to 20 wt percent iodine as Ba(IO 3 ) 2 to provide a fluid mixture and allowing the fluid mixture to harden, said Ba(IO 3 ) 2 comprising said radioactive iodine. An article for solid waste disposal comprises concrete prepared by this method. 10 claims, 2 figures

  19. Contaminant immobilization via microbial activity

    International Nuclear Information System (INIS)

    1991-11-01

    The aim of this study was to search the literature to identify biological techniques that could be applied to the restoration of contaminated groundwaters near uranium milling sites. Through bioremediation it was hypothesized that the hazardous heavy metals could be immobilized in a stable, low-solubility form, thereby halting their progress in the migrating groundwater. Three basic mechanisms were examined: reduction of heavy metals by microbially produced hydrogen sulfide; direct microbial mediated reduction; and biosorption

  20. Developing an Efficient and General Strategy for Immobilization of Small Molecules onto Microarrays Using Isocyanate Chemistry.

    Science.gov (United States)

    Zhu, Chenggang; Zhu, Xiangdong; Landry, James P; Cui, Zhaomeng; Li, Quanfu; Dang, Yongjun; Mi, Lan; Zheng, Fengyun; Fei, Yiyan

    2016-03-16

    Small-molecule microarray (SMM) is an effective platform for identifying lead compounds from large collections of small molecules in drug discovery, and efficient immobilization of molecular compounds is a pre-requisite for the success of such a platform. On an isocyanate functionalized surface, we studied the dependence of immobilization efficiency on chemical residues on molecular compounds, terminal residues on isocyanate functionalized surface, lengths of spacer molecules, and post-printing treatment conditions, and we identified a set of optimized conditions that enable us to immobilize small molecules with significantly improved efficiencies, particularly for those molecules with carboxylic acid residues that are known to have low isocyanate reactivity. We fabricated microarrays of 3375 bioactive compounds on isocyanate functionalized glass slides under these optimized conditions and confirmed that immobilization percentage is over 73%.

  1. Controls of evapotranspiration and CO2 fluxes from scots pine by surface conductance and abiotic factors.

    Directory of Open Access Journals (Sweden)

    Tianshan Zha

    Full Text Available Evapotranspiration (E and CO2 flux (Fc in the growing season of an unusual dry year were measured continuously over a Scots pine forest in eastern Finland, by eddy covariance techniques. The aims were to gain an understanding of their biological and environmental control processes. As a result, there were obvious diurnal and seasonal changes in E, Fc , surface conductance (gc , and decoupling coefficient (Ω, showing similar trends to those in radiation (PAR and vapour pressure deficit (δ. The maximum mean daily values (24-h average for E, Fc , gc , and Ω were 1.78 mmol m(-2 s(-1, -11.18 µmol m(-2 s(-1, 6.27 mm s(-1, and 0.31, respectively, with seasonal averages of 0.71 mmol m(-2 s(-1, -4.61 µmol m(-2 s(-1, 3.3 mm s(-1, and 0.16. E and Fc were controlled by combined biological and environmental variables. There was curvilinear dependence of E on gc and Fc on gc . Among the environmental variables, PAR was the most important factor having a positive linear relationship to E and curvilinear relationship to Fc , while vapour pressure deficit was the most important environmental factor affecting gc . Water use efficiency was slightly higher in the dry season, with mean monthly values ranging from 6.67 to 7.48 μmol CO2 (mmol H2O(-1 and a seasonal average of 7.06 μmol CO2 (μmol H2O(-1. Low Ω and its close positive relationship with gc indicate that evapotranspiration was sensitive to surface conductance. Mid summer drought reduced surface conductance and decoupling coefficient, suggesting a more biotic control of evapotranspiration and a physiological acclimation to dry air. Surface conductance remained low and constant under dry condition, supporting that a constant value of surface constant can be used for modelling transpiration under drought condition.

  2. High quality factor surface Fabry-Perot cavity of acoustic waves

    Science.gov (United States)

    Xu, Yuntao; Fu, Wei; Zou, Chang-ling; Shen, Zhen; Tang, Hong X.

    2018-02-01

    Surface acoustic wave (SAW) resonators are critical components in wireless communications and many sensing applications. They have also recently emerged as a subject of study in quantum acoustics at the single phonon level. Acoustic loss reduction and mode confinement are key performance factors in SAW resonators. Here, we report the design and experimental realization of high quality factor Fabry-Perot SAW resonators formed in between the tapered phononic crystal mirrors patterned on a GaN-on-sapphire material platform. The fabricated SAW resonators are characterized by both an electrical network analyzer and an optical heterodyne vibrometer. We observed standing Rayleigh waves inside the cavity, with an intrinsic quality factor exceeding 1.3 × 104 at ambient conditions.

  3. Fate factors and emission flux estimates for emerging contaminants in surface waters

    Directory of Open Access Journals (Sweden)

    Hoa T. Trinh

    2016-01-01

    Full Text Available Pharmaceuticals, personal care products, hormones, and wastewater products are emerging environmental concerns for manifold reasons, including the potential of some compounds found in these products for endocrine disruption at a very low chronic exposure level. The environmental occurrences and sources of these contaminants in the water, soil, sediment and biota in European nations and the United States are well documented. This work reports a screening-level emission and fate assessment of thirty compounds, listed in the National Reconnaissance of the United States Geological Survey (USGS, 1999–2000 as the most frequently detected organic wastewater contaminants in U.S. streams and rivers. Estimations of the surface water fate factors were based on Level II and Level III multimedia fugacity models for a 1000 km2 model environment, the size of a typical county in the eastern United States. The compounds are categorized into three groups based upon the sensitivity of their predicted surface water fate factors to uncertainties in their physicochemical property values and the landscape parameters. The environmental fate factors, mass distributions, and loss pathways of all of the compounds are strongly affected by their assumed modes of entry into the environment. It is observed that for thirteen of the thirty organic wastewater contaminants most commonly detected in surface waters, conventional treatment strategies may be ineffective for their removal from wastewater effluents. The surface water fate factors predicted by the fugacity models were used in conjunction with the surface water concentrations measured in the USGS reconnaissance to obtain emission flux estimates for the compounds into U.S. streams and rivers. These include estimated fluxes of 6.8 × 10−5 to 0.30 kg/h km2 for the biomarker coprostanol; 1.7 × 10−5 to 6.5 × 10−5 kg/h km2 for the insect repellent N,N-diethyltoluamide; and 4.3 × 10−6 to 3.1 × 10−5 kg/h km2 for

  4. Limonene inhibits streptococcal biofilm formation by targeting surface-associated virulence factors.

    Science.gov (United States)

    Subramenium, Ganapathy Ashwinkumar; Vijayakumar, Karuppiah; Pandian, Shunmugiah Karutha

    2015-08-01

    The present study explores the efficacy of limonene, a cyclic terpene found in the rind of citrus fruits, for antibiofilm potential against species of the genus Streptococcus, which have been deeply studied worldwide owing to their multiple pathogenic efficacy. Limonene showed a concentration-dependent reduction in the biofilm formation of Streptococcus pyogenes (SF370), with minimal biofilm inhibitory concentration (MBIC) of 400 μg ml - 1. Limonene was found to possess about 75-95 % antibiofilm activity against all the pathogens tested, viz. Streptococcus pyogenes (SF370 and 5 clinical isolates), Streptococcus mutans (UA159) and Streptococcus mitis (ATCC 6249) at 400 μg ml - 1 concentration. Microscopic analysis of biofilm architecture revealed a quantitative breach in biofilm formation. Results of a surface-coating assay suggested that the possible mode of action of limonene could be by inhibiting bacterial adhesion to surfaces, thereby preventing the biofilm formation cascade. Susceptibility of limonene-treated Streptococcus pyogenes to healthy human blood goes in unison with gene expression studies in which the mga gene was found to be downregulated. Anti-cariogenic efficacy of limonene against Streptococcus mutans was confirmed, with inhibition of acid production and downregulation of the vicR gene. Downregulation of the covR, mga and vicR genes, which play a critical role in regulating surface-associated proteins in Streptococcus pyogenes and Streptococcus mutans, respectively, is yet further evidence to show that limonene targets surface-associated proteins. The results of physiological assays and gene expression studies clearly show that the surface-associated antagonistic mechanism of limonene also reduces surface-mediated virulence factors.

  5. Factors in the Selection of Surface Disinfectants for Use in a Laboratory Animal Setting

    Science.gov (United States)

    Campagna, Michael V; Faure-Kumar, Emmanuelle; Treger, Janet A; Cushman, Jesse D; Grogan, Tristan R; Kasahara, Noriyuki; Lawson, Gregory W

    2016-01-01

    Because surface disinfectants are an important means of pathogen control within laboratory animal facilities, these products must have an appropriate spectrum of antimicrobial activity. However, many other factors must also be considered, including effects on human health, environmental safety, and animal behavior. Aqueous solutions of sodium hypochlorite often are considered to be the ‘gold standard’ for surface disinfection, but these products can be corrosive, caustic, and aversive in odor. This study was designed to identify disinfectants that are as effective as hypochlorite solutions but more acceptable for use in a laboratory animal setting. An antiviral disinfectant-efficacy assay was developed by using viral vectors that expressed green fluorescence protein as surrogates for wild-type viruses of concern in laboratory animals. Efficacy testing revealed that most of the products were highly effective when used against viral vectors in suspension. However, when the disinfectants were challenged by buffering virus in protein or drying virus on nonporous surfaces, the hypochlorite and peroxymonosulfate products performed the best. Review of safety data sheets for the agents indicated that a peroxide-based product was considerably safer than the other products tested and that the pH of most products was not conducive to disposal down a drain. Behavioral testing of Swiss Webster, C57Bl/6, and BALB/c mice showed that the hypochlorite- and peroxide-based products were clearly aversive, given that the mice consistently avoided these products. All of these factors must be considered when choosing the appropriate disinfectant. PMID:27025810

  6. [Factors related to the influence on congenital malformation of body surface in Shaanxi province].

    Science.gov (United States)

    Ying, J; Lei, Q; Qin, B W; Qu, P F; Lei, F L; Dang, S N; Zeng, L X; Yan, H

    2016-12-10

    Objective: To investigate the influencing factors on congenital malformation of body surface. Methods: Multi-stage stratified random sampling method was adopted. A questionnaire survey was conducted among the childbearing aged women who experienced more than 28 weeks pregnancy or having definite pregnancy outcomes in Shaanxi, during 2010-2013. Results: Results from the logistic regression model showed that factors as: intrahepatic cholestasis of pregnancy ( OR =21.76, 95 %CI : 4.46-106.25), histories with abnormal pregnancy or reproduction ( OR =11.88, 95 %CI : 9.14-15.45), having birth defects in the family ( OR =6.15, 95 %CI : 2.66-14.23), being twins ( OR =5.74, 95 % CI : 3.34-9.86), being worker (compared with others, OR =2.47, 95 % CI : 1.30-4.68) or farmer (compared with others, OR =1.91, 95 %CI : 1.14-3.20), less than 4 times of antenatal care (compared with >7, OR =1.84, 95 % CI : 1.28-2.64), occupational exposure to related risks during pregnancy ( OR =1.74, 95 % CI : 1.26-2.42), mother's hometown was from mid-part of Shaanxi (compared with northern Shaanxi, OR =1.65, 95 %CI : 1.20-2.28), mother's native residence was from the rural areas ( OR =1.75, 95 % CI : 1.13-2.71), drug use ( OR =1.64, 95 % CI : 1.26-2.13) etc . were risk factors for congenital malformation of body surface. Iron supplement during pregnancy ( OR =0.46, 95 %CI : 0.21-0.99) appeared as protective factor for congenital malformation of body surface. Conclusion: The following factors seemed to be of risk for congenital malformation of body surface, including: mother's native area was from the middle part of Shaanxi and living in rural area, being worker or farmer, histories of abnormal pregnancy and reproduction, history of birth defects in the family, being twins, with antenatal care less than 4 times, occupational exposure to dangerous materials, drug use, intrahepatic cholestasis during pregnancy etc .

  7. The evaluation for usefulness of the custom made immobilization device for the anteroperitoneal resection patients with rectal cancer

    International Nuclear Information System (INIS)

    Yang, Oh Nam; Lee, Woo Seok; Hong, Tae Kyun; Jo, Young Pih; Yun, Hwa Ryong; Kim, Jung Man

    2003-01-01

    Patient immobilization is essential factor for successful radiation therapy and major problem is reproducibility to maintain patient position during total radiation therapy period. Purpose of this study is evaluation for usefulness of the custom made immobilization device for the anteroperitoneal resection patients with rectal cancer. The object of this study were patients who underwent anteroperitoneal resection and undergo radiation therapy at present with rectal cancer. We made immobilization device for patient individually and analyzed its set up reproducibility, patient position deviation and errors. There was 5 mm-10 mm deviation in patient position without individual immobilization device, but we improved the deviation within few mm limitation with individual immobilization device. Custom made immobilization device was very helpful for anteroperitoneal resection patient with rectal cancer. We improved the patient position deviation within few mm limitation, shorten the set up time and we could give the comfort to patients.

  8. Covalent immobilization of molecularly imprinted polymer nanoparticles using an epoxy silane.

    Science.gov (United States)

    Kamra, Tripta; Chaudhary, Shilpi; Xu, Changgang; Johansson, Niclas; Montelius, Lars; Schnadt, Joachim; Ye, Lei

    2015-05-01

    Molecularly imprinted polymers (MIPs) can be used as antibody mimics to develop robust chemical sensors. One challenging problem in using MIPs for sensor development is the lack of reliable conjugation chemistry that allows MIPs to be fixed on transducer surface. In this work, we study the use of epoxy silane to immobilize MIP nanoparticles on model transducer surfaces without impairing the function of the immobilized nanoparticles. The MIP nanoparticles with a core-shell structure have selective molecular binding sites in the core and multiple amino groups in the shell. The model transducer surface is functionalized with a self-assembled monolayer of epoxy silane, which reacts with the core-shell MIP particles to enable straightforward immobilization. The whole process is characterized by studying the treated surfaces after each preparation step using atomic force microscopy, scanning electron microscopy, fluorescence microscopy, contact angle measurements and X-ray photoelectron spectroscopy. The microscopy results show that the MIP particles are immobilized uniformly on surface. The photoelectron spectroscopy results further confirm the action of each functionalization step. The molecular selectivity of the MIP-functionalized surface is verified by radioligand binding analysis. The particle immobilization approach described here has a general applicability for constructing selective chemical sensors in different formats. Copyright © 2015 The Authors. Published by Elsevier Inc. All rights reserved.

  9. Mechanical Q-factor measurements on a test mass with a structured surface

    Energy Technology Data Exchange (ETDEWEB)

    Nawrodt, R [Institut fuer Festkoerperphysik, Friedrich-Schiller-Universitaet Jena, Helmholtzweg 5, D-07743 Jena (Germany); Zimmer, A [Institut fuer Festkoerperphysik, Friedrich-Schiller-Universitaet Jena, Helmholtzweg 5, D-07743 Jena (Germany); Koettig, T [Institut fuer Festkoerperphysik, Friedrich-Schiller-Universitaet Jena, Helmholtzweg 5, D-07743 Jena (Germany); Clausnitzer, T [Institut fuer Angewandte Physik, Friedrich-Schiller-Universitaet Jena, Max-Wien-Platz 1, D-07743 Jena (Germany); Bunkowski, A [Max-Planck-Institut fuer Gravitationsphysik (Albert-Einstein-Institut) and Institut fuer Gravitationsphysik, Leibniz Universitaet Hannover, Callinstr. 38, D-30167 Hannover (Germany); Kley, E B [Institut fuer Angewandte Physik, Friedrich-Schiller-Universitaet Jena, Max-Wien-Platz 1, D-07743 Jena (Germany); Schnabel, R [Max-Planck-Institut fuer Gravitationsphysik (Albert-Einstein-Institut) and Institut fuer Gravitationsphysik, Leibniz Universitaet Hannover, Callinstr. 38, D-30167 Hannover (Germany); Danzmann, K [Max-Planck-Institut fuer Gravitationsphysik (Albert-Einstein-Institut) and Institut fuer Gravitationsphysik, Leibniz Universitaet Hannover, Callinstr. 38, D-30167 Hannover (Germany); Nietzsche, S [Institut fuer Festkoerperphysik, Friedrich-Schiller-Universitaet Jena, Helmholtzweg 5, D-07743 Jena (Germany); Vodel, W [Institut fuer Festkoerperphysik, Friedrich-Schiller-Universitaet Jena, Helmholtzweg 5, D-07743 Jena (Germany); Tuennermann, A [Institut fuer Angewandte Physik, Friedrich-Schiller-Universitaet Jena, Max-Wien-Platz 1, D-07743 Jena (Germany); Seidel, P [Institut fuer Festkoerperphysik, Friedrich-Schiller-Universitaet Jena, Helmholtzweg 5, D-07743 Jena (Germany)

    2007-07-15

    We present mechanical Q-factors (quality factors) of a crystalline quartz test mass with a nano-structured surface, measured in the temperature regime from 5 to 300 K. The nano-structure was a grating with a period of 2 {mu}m and a depth of about 0.1 {mu}m. Comparative measurements were performed on the plain substrate and on the structured test mass with different numbers of SiO{sub 2}/Ta{sub 2}O{sub 5} coating layers. The measurements at different stages of the test mass fabrication process show that the surface distortion induced by the nanostructure does not severely lower the mechanical Q-factor of the substrate. Damping due to a multi-layer coating stack was found to be orders of magnitude higher. The results provide vital information concerning the potential usage of low-thermal noise nano-structured test masses in future generations of high-precision laser interferometers and in current attempts to measure quantum effects of macroscopic mirror oscillators.

  10. Herbaceous plants as filters: Immobilization of particulates along urban street corridors

    International Nuclear Information System (INIS)

    Weber, Frauke; Kowarik, Ingo; Säumel, Ina

    2014-01-01

    Among air pollutants, particulate matter (PM) is considered to be the most serious threat to human health. Plants provide ecosystem services in urban areas, including reducing levels of PM by providing a surface for deposition and immobilization. While previous studies have mostly addressed woody species, we focus on herbaceous roadside vegetation and assess the role of species traits such as leaf surface roughness or hairiness for the immobilization of PM. We found that PM deposition patterns on plant surfaces reflect site-specific traffic densities and that strong differences in particulate deposition are present among species. The amount of immobilized PM differed according to particle type and size and was related to specific plant species traits. Our study suggests that herbaceous vegetation immobilizes a significant amount of the air pollutants relevant to human health and that increasing biodiversity of roadside vegetation supports air filtration and thus healthier conditions along street corridors. -- Highlights: • We assessed PM immobilization by common urban herbaceous roadside species. • PM deposition was related to traffic density and plant species traits. • Amount of PM deposited differed according to particle type and size. • Increasing biodiversity of roadside vegetation supports air filtration. -- Herbaceous urban roadside vegetation immobilizes particulate matter relevant to human health, thus supporting healthier conditions next to busy roads

  11. Shoe and Field Surface Risk Factors for Acute Lower Extremity Injuries Among Female Youth Soccer Players.

    Science.gov (United States)

    OʼKane, John W; Gray, Kristen E; Levy, Marni R; Neradilek, Moni; Tencer, Allan F; Polissar, Nayak L; Schiff, Melissa A

    2016-05-01

    To describe acute lower extremity injuries and evaluate extrinsic risk factors in female youth soccer. Nested case-control study. Youth soccer clubs in Seattle, WA. Female soccer players (n = 351) ages 11 to 15 years randomly selected from 4 soccer clubs from which 83% of their players were enrolled with complete follow-up for 92% of players. Injured players were interviewed regarding injury, field surface, shoe type, and position. Uninjured controls, matched on game or practice session, were also interviewed. The association between risk factors and acute lower extremity injury using logistic regression to estimate odds ratios (OR) and 95% confidence intervals (CI). One hundred seventy-three acute lower extremity injuries occurred involving primarily the ankle (39.3%), knee (24.9%), and thigh (11.0%). Over half (52.9%) recovered within 1 week, whereas 30.2% lasted beyond 2 weeks. During practices, those injured were approximately 3-fold (OR, 2.83; 95% CI, 1.49-5.31) more likely to play on grass than artificial turf and 2.4-fold (95% CI, 1.03-5.96) more likely to wear cleats on grass than other shoe and surface combinations. During games, injured players were 89% (95% CI, 1.03-4.17) more likely to play defender compared with forward. Half of the acute lower extremity injuries affected the ankle or knee. Grass surface and wearing cleats on grass increased training injuries. The majority, 64%, of female youth soccer players' acute injuries involve the ankle and knee and injury prevention strategies in this age group should target these areas. When considering playing surfaces for training, communities and soccer organizations should consider the third-generation artificial turf a safe alternative to grass.

  12. A simple gold nanoparticle-mediated immobilization method to fabricate highly homogeneous DNA microarrays having higher capacities than those prepared by using conventional techniques

    International Nuclear Information System (INIS)

    Jung, Cheulhee; Mun, Hyo Young; Li, Taihua; Park, Hyun Gyu

    2009-01-01

    A simple, highly efficient immobilization method to fabricate DNA microarrays, that utilizes gold nanoparticles as the mediator, has been developed. The fabrication method begins with electrostatic attachment of amine-modified DNA to gold nanoparticles. The resulting gold-DNA complexes are immobilized on conventional amine or aldehyde functionalized glass slides. By employing gold nanoparticles as the immobilization mediator, implementation of this procedure yields highly homogeneous microarrays that have higher binding capacities than those produced by conventional methods. This outcome is due to the increased three-dimensional immobilization surface provided by the gold nanoparticles as well as the intrinsic effects of gold on emission properties. This novel immobilization strategy gives microarrays that produce more intense hybridization signals for the complementary DNA. Furthermore, the silver enhancement technique, made possible only in the case of immobilized gold nanoparticles on the microarrays, enables simple monitoring of the integrity of the immobilized DNA probe.

  13. Unpacking the enabling factors for hand, cord and birth-surface hygiene in Zanzibar maternity units.

    Science.gov (United States)

    Gon, Giorgia; Ali, Said M; Towriss, Catriona; Kahabuka, Catherine; Ali, Ali O; Cavill, Sue; Dahoma, Mohammed; Faulkner, Sally; Haji, Haji S; Kabole, Ibrahim; Morrison, Emma; Said, Rukaiya M; Tajo, Amour; Velleman, Yael; Woodd, Susannah L; Graham, And Wendy J

    2017-10-01

    Recent national surveys in The United Republic of Tanzania have revealed poor standards of hygiene at birth in facilities. As more women opt for institutional delivery, improving basic hygiene becomes an essential part of preventative strategies for reducing puerperal and newborn sepsis. Our collaborative research in Zanzibar provides an in-depth picture of the state of hygiene on maternity wards to inform action. Hygiene was assessed in 2014 across all 37 facilities with a maternity unit in Zanzibar. We used a mixed methods approach, including structured and semi-structured interviews, and environmental microbiology. Data were analysed according to the WHO 'cleans' framework, focusing on the fundamental practices for prevention of newborn and maternal sepsis. For each 'clean' we explored the following enabling factors: knowledge, infrastructure (including equipment), staffing levels and policies. Composite indices were constructed for the enabling factors of the 'cleans' from the quantitative data: clean hands, cord cutting, and birth surface. Results from the qualitative tools were used to complement this information.Only 49% of facilities had the 'infrastructural' requirements to enable 'clean hands', with the availability of constant running water particularly lacking. Less than half (46%) of facilities met the 'knowledge' requirements for ensuring a 'clean delivery surface'; six out of seven facilities had birthing surfaces that tested positive for multiple potential pathogens. Almost two thirds of facilities met the 'infrastructure (equipment) requirement' for 'clean cord'; however, disposable cord clamps being frequently out of stock, often resulted in the use of non-sterile thread made of fabric. This mixed methods approach, and the analytical framework based on the WHO 'cleans' and the enabling factors, yielded practical information of direct relevance to action at local and ministerial levels. The same approach could be applied to collect and analyse data

  14. Trends in interfacial design for surface plasmon resonance based immunoassays

    Energy Technology Data Exchange (ETDEWEB)

    Shankaran, Dhesingh Ravi [Art, Science and Technology Center for Cooperative Research, Kyushu University, Kasuga-shi, Fukuoka, 816-8580 (Japan); Miura, Norio [Art, Science and Technology Center for Cooperative Research, Kyushu University, Kasuga-shi, Fukuoka, 816-8580 (Japan)

    2007-12-07

    Immunosensors based on surface plasmon resonance (SPR) have become a promising tool in sensor technology for biomedical, food, environmental, industrial and homeland security applications. SPR is a surface sensitive optical technique, suitable for real-time and label-free analysis of biorecognition events at functional transducer surfaces. Fabrication of highly active and robust sensing surfaces is an important part in immunoassays because the quality, quantity, chemistry and topography of the interfacial biomembranes play a major role in immunosensor performance. Eventually, a variety of immobilization methods such as physical adsorption, covalent coupling, Langmuir-Blodgett film, polymer thin film, self-assembly, sol-gel, etc, have been introduced over the years for the immobilization of biomolecules (antibody or antigen) on the transducer surfaces. The selection of an immobilization method for an immunoassay is governed by several factors such as nature and stability of the biomolecules, target analyte, application, detection principle, mode of signal transduction, matrix complexity, etc. This paper provides an overview of the various surface modification methods for SPR based immunosensor fabrication. The preparation, structure and application of different functional interfacial surfaces have been discussed along with a brief introduction to the SPR technology, biomolecules and detection principles. (review article)

  15. Trends in interfacial design for surface plasmon resonance based immunoassays

    International Nuclear Information System (INIS)

    Shankaran, Dhesingh Ravi; Miura, Norio

    2007-01-01

    Immunosensors based on surface plasmon resonance (SPR) have become a promising tool in sensor technology for biomedical, food, environmental, industrial and homeland security applications. SPR is a surface sensitive optical technique, suitable for real-time and label-free analysis of biorecognition events at functional transducer surfaces. Fabrication of highly active and robust sensing surfaces is an important part in immunoassays because the quality, quantity, chemistry and topography of the interfacial biomembranes play a major role in immunosensor performance. Eventually, a variety of immobilization methods such as physical adsorption, covalent coupling, Langmuir-Blodgett film, polymer thin film, self-assembly, sol-gel, etc, have been introduced over the years for the immobilization of biomolecules (antibody or antigen) on the transducer surfaces. The selection of an immobilization method for an immunoassay is governed by several factors such as nature and stability of the biomolecules, target analyte, application, detection principle, mode of signal transduction, matrix complexity, etc. This paper provides an overview of the various surface modification methods for SPR based immunosensor fabrication. The preparation, structure and application of different functional interfacial surfaces have been discussed along with a brief introduction to the SPR technology, biomolecules and detection principles. (review article)

  16. Immobilization of radioactive strontium in contaminated soils by phosphate treatment

    International Nuclear Information System (INIS)

    Kim, K.H.; Ammons, J.T.

    1990-01-01

    The feasibility of in situ phosphate- and metal- (calcium, aluminum, and iron) solution treatment for 90 Sr immobilization was investigated. Batch and column experiments were performed to find optimum conditions for coprecipitation of 90 Sr with Ca-, Al-, and Fe-phosphate compounds in contaminated soils. Separate columns were packed with artificially 85 Sr-contaminated acid soil as well as 90 Sr-contaminated soil from the Oak Ridge Reservation. After metal-phosphate treatment, the columns were then leached successively with either tapwater or 0.001 M CaCl 2 solution. Most of the 85 Sr coprecipitated with the metal phosphate compounds. Immobilization of 85 Sr and 90 Sr was affected by such factors as solution pH, metal and phosphate concentration, metal-to-phosphate ratio, and soil characteristics. Equilibration time after treatments also affected 85 Sr immobilization. Many technology aspects still need to be investigated before field applications are feasible, but these experiments indicate that phosphate-based in situ immobilization should prevent groundwater contamination and will be useful as a treatment technology for 90 Sr-contaminated sites. 15 refs., 3 figs., 1 tab

  17. Pellet formation of zygomycetes and immobilization of yeast.

    Science.gov (United States)

    Nyman, Jonas; Lacintra, Michael G; Westman, Johan O; Berglin, Mattias; Lundin, Magnus; Lennartsson, Patrik R; Taherzadeh, Mohammad J

    2013-06-25

    Pelleted growth provides many advantages for filamentous fungi, including decreased broth viscosity, improved aeration, stirring, and heat transfer. Thus, the factors influencing the probability of pellet formation of Rhizopus sp. in a defined medium was investigated using a multifactorial experimental design. Temperature, agitation intensity, Ca(2+)-concentration, pH, and solid cellulose particles, each had a significant effect on pelletization. Tween 80, spore concentration, and liquid volume were not found to have a significant effect. All of the effects were additive; no interactions were significant. The results were used to create a simple defined medium inducing pelletization, which was used for immobilization of a flocculating strain of Saccharomyces cerevisiae in the zygomycetes pellets. A flor-forming S. cerevisiae strain was also immobilized, while a non-flocculating strain colonized the pellets but was not immobilized. No adverse effects were detected as a result of the close proximity between the filamentous fungus and the yeast, which potentially allows for co-fermentation with S. cerevisiae immobilized in pellets of zygomycetes. Copyright © 2013 Elsevier B.V. All rights reserved.

  18. Silk-Cocoon Matrix Immobilized Lipase Catalyzed Transesterification of Sunflower Oil for Production of Biodiesel

    Directory of Open Access Journals (Sweden)

    Sushovan Chatterjee

    2014-01-01

    Full Text Available Biodiesel from sunflower oil using lipase chemically immobilized on silk-cocoon matrix in a packed-bed bioreactor was investigated. The immobilization was demonstrated by field-emission scanning electron microscopy and activity study. The lipase loading was 738.74 U (~0.01 g lipase powder/g-lipase-immobilized matrix. The Km (Michaelis-Menten constant of the free and the immobilized lipase was 451.26 μM and 257.26 μM, respectively. Low Km value of the immobilized lipase is attributed to the hydrophobic nature of the matrix that facilitated the substrate diffusion to the enzyme surface. The biodiesel yield of 81.62% was obtained at 48 hours reaction time, 6 : 1 methanol : oil ratio (v/v, and 30°C. The immobilized lipase showed high operational stability at 30°C. The substrate conversion was only marginally decreased till third cycle (each of 48 hours duration of the reaction since less than even 5% of the original activity was decreased in each of the second and third cycle. The findings demonstrated the potential of the silk-cocoon as lipase immobilization matrix for industrial production of biodiesel.

  19. Preliminary assessment of modified borosilicate glasses for chromium and ruthenium immobilization

    Energy Technology Data Exchange (ETDEWEB)

    Farid, Osama M. [Reactors Department, Nuclear Research Center, Atomic Energy Authority of Egypt, P.O. 13759, Inshas, Cairo (Egypt); Centre of Nuclear Engineering (CNE), Department of Materials, Imperial College London, London, SW7 2BP (United Kingdom); Abdel Rahman, R.O., E-mail: alaarehab@yahoo.com [Hot Laboratory Center, Atomic Energy Authority of Egypt, P.O. 13759, Inshas, Cairo (Egypt)

    2017-01-15

    The feasibility of using modified alkali borosilicate glasses for ruthenium and chromium immobilization is preliminary assessed by investigating the immobilization system structure under normal conditions. Within this context, reference alkali borosilicate, and simulated Magnox-modified glasses were prepared and studied. The results indicate that ruthenium is immobilized in the vitreous structure as encapsulated RuO{sub 2} crystallites that act as seeds for heterogeneous nucleation of other crystalline phases. The presence of Zn, as modifier, has contributed to chromium immobilization in zincochromite spinel structure, whereas Ca is accommodated in the vitreous structure. Immobilization performance was evaluated by conducting conservative static leach test and studying the leached glass. Leached glass morphology was altered, where near surface reference glass is leached over 400 nm and simulated Magnox-modified sample is altered over 300 nm. Normalized release rates are within normal range for borosilicate material. For simulated Magnox-modified sample, Ca and alkali structural element, i.e. Na and Li, are leached via ion-exchange reaction. The ion-exchanged fraction equals 1.06 × 10{sup −8} mol/m{sup 2} s and chromium has slightly lower normalized release rate value than ruthenium. - Highlights: • The presence of modifiers and waste oxides led to localized de-vitrification. • Ruthenium is encapsulated within the vitreous glass network as RuO{sub 2} crystals. • Chromium is immobilized within the zincochromite spinel structure. • Pitting and cracks induced by leaching did not affect the immobilization performance.

  20. Antimicrobial Properties of Lysosomal Enzymes Immobilized on NH₂Functionalized Silica-Encapsulated Magnetite Nanoparticles.

    Science.gov (United States)

    Bang, Seung Hyuck; Sekhon, Simranjeet Singh; Cho, Sung-Jin; Kim, So Jeong; Le, Thai-Hoang; Kim, Pil; Ahn, Ji-Young; Kim, Yang-Hoon; Min, Jiho

    2016-01-01

    The immobilization efficiency, antimicrobial activity and recovery of lysosomal enzymes on NH2 functionalized magnetite nanoparticles have been studied under various conditions. The immobi- lization efficiency depends upon the ratio of the amount of enzyme and magnetite and it shows an increase with magnetite concentration which is due to the presence of amine group at the magnetite surface that leads to a strong attraction. The optimized reaction time to immobilize the lysosomal enzymes on magnetite was determined by using a rolling method. The immobilization efficiency increases with reaction time and reached a plateau after 5 minutes and then remained constant for 10 minutes. However, after 30 minutes the immobilization efficiency decreased to 85%, which is due to the weaker electrostatic interactions between magnetite and detached lysosomal enzymes. The recovery and stability of immobilized lysosomal enzymes has also been studied. The antimicrobial activity was almost 100% but it decreased upon reuse and no activity was observed after its reuse for seven times. The storage stability of lysosomal enzymes as an antimicrobial agent was about 88%, which decreased to 53% after one day and all activity of immobilized lysosomal enzymes was maintained after five days. Thus, the lysosomal enzymes immobilized on magnetite nanoparticles could potentially be used as antimicrobial agents to remove bacteria.

  1. Evaluation of Styrene-Divinylbenzene Beads as a Support to Immobilize Lipases

    Directory of Open Access Journals (Sweden)

    Cristina Garcia-Galan

    2014-06-01

    Full Text Available A commercial and very hydrophobic styrene-divinylbenzene matrix, MCI GEL® CHP20P, has been compared to octyl-Sepharose® beads as support to immobilize three different enzymes: lipases from Thermomyces lanuginosus (TLL and from Rhizomucor miehie (RML and Lecitase® Ultra, a commercial artificial phospholipase. The immobilization mechanism on both supports was similar: interfacial activation of the enzymes versus the hydrophobic surface of the supports. Immobilization rate and loading capacity is much higher using MCI GEL® CHP20P compared to octyl-Sepharose® (87.2 mg protein/g of support using TLL, 310 mg/g using RML and 180 mg/g using Lecitase® Ultra. The thermal stability of all new preparations is much lower than that of the standard octyl-Sepharose® immobilized preparations, while the opposite occurs when the inactivations were performed in the presence of organic co-solvents. Regarding the hydrolytic activities, the results were strongly dependent on the substrate and pH of measurement. Octyl-Sepharose® immobilized enzymes were more active versus p-NPB than the enzymes immobilized on MCI GEL® CHP20P, while RML became 700-fold less active versus methyl phenylacetate. Thus, the immobilization of a lipase on this matrix needs to be empirically evaluated, since it may present very positive effects in some cases while in other cases it may have very negative ones.

  2. Management of primary anterior shoulder dislocations using immobilization.

    Science.gov (United States)

    Smith, Brent I; Bliven, Kellie C Huxel; Morway, Genoveffa R; Hurbanek, Jason G

    2015-05-01

    identified 2083 articles. A total of 9 studies met all of the criteria and were included in this review. In most of the studies, age was a risk factor for recurrence. Patients less than 30 years of age were more likely to sustain a recurrent dislocation than patients more than 30 years of age. In 5 studies (n = 1215), researchers found no difference in recurrence of shoulder dislocation when immobilized in internal rotation (IR) for less than 1 week (41%, 40 of 97) compared with more than 3 weeks (37%, 34 of 93) in patients less than 30 years of age (P = .52). Authors of 3 studies (n = 289) compared the effect of immobilization in IR versus external rotation (ER), and whereas they found no statistical difference, a trend appeared toward reduced recurrence rates in ER but not IR (P = .07). The rate of recurrent dislocation was 40% (25 of 63) in patients treated with IR sling immobilization and 25% (22 of 88) in patients immobilized in ER. Overall, the investigators found that younger age (immobilization for more than 1 week did not improve recurrence rates, and an apparent trend existed toward decreased recurrence rates with ER rather than IR. According to the review and meta-analysis by Paterson et al, the level of evidence for recommendations regarding optimal duration and position of immobilization to reduce the risk of recurrent dislocation was therapeutic level II. This level of evidence was appropriate because the review included only prospective studies of level I or II and a minimum follow-up of 1 year.

  3. Surface water acidification and critical loads: exploring the F-factor

    Directory of Open Access Journals (Sweden)

    K. Bishop

    2009-11-01

    Full Text Available As acid deposition decreases, uncertainties in methods for calculating critical loads become more important when judgements have to be made about whether or not further emission reductions are needed. An important aspect of one type of model that has been used to calculate surface water critical loads is the empirical F-factor which estimates the degree to which acid deposition is neutralised before it reaches a lake at any particular point in time relative to the pre-industrial, steady-state water chemistry conditions.

    In this paper we will examine how well the empirical F-functions are able to estimate pre-industrial lake chemistry as lake chemistry changes during different phases of acidification and recovery. To accomplish this, we use the dynamic, process-oriented biogeochemical model SAFE to generate a plausible time series of annual runoff chemistry for ca. 140 Swedish catchments between 1800 and 2100. These annual hydrochemistry data are then used to generate empirical F-factors that are compared to the "actual" F-factor seen in the SAFE data for each lake and year in the time series. The dynamics of the F-factor as catchments acidify, and then recover are not widely recognised.

    Our results suggest that the F-factor approach worked best during the acidification phase when soil processes buffer incoming acidity. However, the empirical functions for estimating F from contemporary lake chemistry are not well suited to the recovery phase when the F-factor turns negative due to recovery processes in the soil. This happens when acid deposition has depleted the soil store of BC, and then acid deposition declines, reducing the leaching of base cations to levels below those in the pre-industrial era. An estimate of critical load from water chemistry during recovery and empirical F functions would therefore result in critical loads that are too low. Therefore, the empirical estimates of the F-factor are a significant source of

  4. Dependence of protein binding capacity of dimethylamino-γ-butyric-acid (DMGABA)-immobilized porous membrane on composition of solvent used for DMGABA immobilization

    International Nuclear Information System (INIS)

    Iwanade, Akio; Umeno, Daisuke; Saito, Kyoichi; Sugo, Takanobu

    2013-01-01

    Dimethylamino-γ-butyric acid (DMGABA) as an ampholite was reacted with the epoxy group of the poly-glycidyl methacrylate chain grafted onto the pore surface of a porous hollow-fiber polyethylene membrane by radiation-induced graft polymerization. DMGABA was dissolved in a mixture of dioxane and water at various dioxane volume fractions, defined by dividing the dioxane volume by the total volume. The equilibrium binding capacity (EBC) of the DMGABA-immobilized porous hollow-fiber membrane for lysozyme was evaluated in the permeation mode. The EBC was varied from a 1/50-fold monolayer binding capacity to a 10-fold monolayer binding capacity by controlling the composition of the solvent used for DMGABA immobilization and the molar conversion of the epoxy group into the DMGABA group. - Highlights: ► A DMGABA membrane was immobilized by irradiation induced graft polymerization. ► The DMGABA was immobilized in a mixture of dioxane and water of various compositions. ► Lysozyme adsorptivity of DMGABA-immobilized membranes evaluated in the permeation mode. ► The composition of the DMGABA immobilized solvent can control adsorptivity

  5. Bioavailable nitrate detection in water by an immobilized luminescent cyanobacterial reporter strain.

    Science.gov (United States)

    Mbeunkui, F; Richaud, C; Etienne, A-L; Schmid, R D; Bachmann, T T

    2002-11-01

    Cyanobacteria are a major group of photosynthetic bacteria that can accumulate in surface water as so-called "blooms" in response to environmental factors such as temperature, light and certain nutrients such as N, P, and Fe. Some species of cyanobacteria produce toxins, causing a considerable danger for human and livestock health. As a consequence, monitoring of bloom formation and toxin production of drinking water supplies has become a major concern. To enable prediction and monitoring of cyanobacterial blooms, tools to detect nutrient bioavailability in water would be advantageous. A whole-cell biosensor was developed for monitoring nitrate (NO(3-)) bioavailability in aquatic ecosystems using the recombinant bioluminescent cyanobacterial strain Synechocystis PCC 6803 harboring an insertion of a luxAB-kmr fusion with nblA1 in its chromosomal DNA, leading to PnblA::luxAB-kmr. This reporter strain was designated N1LuxKm. Cells were immobilized in microtiter plates and showed a dose-dependent response to nitrate deprivation. The resultant CyanoSensor could detect nitrate in the 4-100 micro M concentration range after a sample incubation time of 10 h under continuous illumination (50 micro E m(-2) s(-1)). The optimal temperature for sensor operation was 29 degrees C and the immobilized biosensor could be stored at 4 degrees C in dark for about 1 month without significant loss of sensitivity.

  6. Development of new Z-factors for the evaluation of the circumferential surface crack in nuclear pipes

    International Nuclear Information System (INIS)

    Choi, Y.H.; Chung, Y.K.; Park, Y.W.; Lee, J.B.

    1997-01-01

    The purpose of this study is to develop new Z-factors to evaluate the behavior of a circumferential surface crack in nuclear pipe. Z-factor is a load multiplier used in the Z-factor method, which is one of the ASME Code Sec. XI's recommendations for the estimation of a surface crack in nuclear pipe. It has been reported that the load carrying capacities predicted from the current ASME Code Z-factors, are not well in agreement with the experimental results for nuclear pipes with a surface crack. In this study, new Z-factors for ferritic base metal, ferritic submerged arc welding (SAW) weld metal, austenitic base metal, and austenitic SAW weld metal are obtained by use of the surface crack for thin pipe (SC.TNP) method based on GE/EPRI method. The desirability of both the SC.TNP method and the new Z-factors is examined using the results from 48 pipe fracture experiments for nuclear pipes with a circumferential surface crack. The results show that the SC.TNP method is good for describing the circumferential surface crack behavior and the new Z-factors are well in agreement with the measured Z-factors for both ferritic and austenitic pipes. (orig.)

  7. Development of phage/antibody immobilized magnetostrictive biosensors

    Science.gov (United States)

    Fu, Liling

    There is an urgent need for biosensors that are able to detect and quantify the presence of a small amount of pathogens in a real-time manner accurately and quickly to guide prevention efforts and assay food and water quality. Acoustic wave (AW) devices, whose performance is defined by mass sensitivity (Sm) and quality factor (Q value), have been extensively studied as high performance biosensor platforms. However, current AW devices still face some challenges such as the difficulty to be employed in liquid and low Q value in practical applications. The objective of this research is to develop magnetostrictive sensors which include milli/microcantilever type (MSMC) and particle type (MSP). Compared to other AW devices, MSMC exhibits the following advantages: (1) wireless/remote driving and sensing; (2) easy to fabricate; (3) works well in liquid; (4) exhibits a high Q value (> 500 in air). The fundamental study of the damping effect on MSMCs from the surrounding media including air and liquids were conducted to improve the Q value of MSMCs. The experiment results show that the Q value is dependent on the properties of surrounding media (e.g. viscosity, density), the geometry of the MSMCs, and the harmonic mode on the resonance behavior of MSMCs, etc. The phage-coated MSMC has high specificity and sensitivity even while used in water with a low concentration of targeted bacteria. Two currently developed phages, JRB7 and E2, respectively respond to Bacillus anthracis spores and Salmonella typhimurium, were employed as bio-recognition elements in this research. The phage-immobilized MSMC biosensors exhibited high performance and detection of limit was 5 x 104 cfu/ml for the MSMC in size of 1.4 x 0.8 x 0.035 mm. The MSMC-based biosensors were indicated as a very potential method for in-situ monitoring of the biological quality in water. The MSP combine antibody was used to detect Staphylococcus aureus in this experiment. The interface between MSPs and antibody was

  8. A novel method for immobilization of Co tetraphenylporphyrins on P(4VP-co-St)/SiO 2: Efficient catalysts for aerobic oxidation of ethylbenzenes

    Science.gov (United States)

    Wang, Ruixin; Gao, Baojiao; Jiao, Weizhou

    2009-01-01

    In this paper, poly(4vinylpyridine-co-styrene) (P(4VP-co-St)) was grafted on silica gel particles in the manner of "grafting from", and the grafting particle P(4VP-co-St)/SiO 2 was gained. The grafting particle P(4VP-co-St)/SiO 2 is a novel kind of supports for immobilizing metalloporphyrin catalysts. Then, the immobilization of cobalt tetraphenylporphyrin (CoTPP) on the supports P(4VP-co-St)/SiO 2 was carried out via the axial coordination reaction between CoTPP and the pyridine groups of the grafted P(4VP-co-St), resulting in the heterogenised catalysts CoTPP-P(4VP-co-St)/SiO 2. The synthesized catalysts were characterized by FTIR and the axial coordination process between CoTPP and the grafted P(4VP-co-St) was confirmed by UV-vis. The effects of various factors on the immobilization reaction of CoTPP were studied in detail. Finally, the catalytic performance of CoTPP-P(4VP-co-St)/SiO 2 in the catalytic oxidation process of ethyl benzene was investigated. The experimental results show that the axial coordination reaction is a very easy and novel method for favorably immobilizing CoTPP onto the P(4VP-co-St)/SiO 2 surfaces. During the immobilization process of CoTPP on P(4VP-co-St)/SiO 2, the most bonding amount of CoTPP (0.19 g/g) is obtained under the lower temperature (5 °C) and the higher concentration of CoTPP(6.0 mg/ml) lasting 4 h. Moreover, the supported catalyst CoTPP-P(4VP-co-St)/SiO 2 can effectively activate the dioxygen, and obviously catalyze the transform of ethylbenzene into acetophenone. So it exhibits the fine catalytic activity.

  9. Surface-based morphometry reveals the neuroanatomical basis of the five-factor model of personality.

    Science.gov (United States)

    Riccelli, Roberta; Toschi, Nicola; Nigro, Salvatore; Terracciano, Antonio; Passamonti, Luca

    2017-04-01

    The five-factor model (FFM) is a widely used taxonomy of human personality; yet its neuro anatomical basis remains unclear. This is partly because past associations between gray-matter volume and FFM were driven by different surface-based morphometry (SBM) indices (i.e. cortical thickness, surface area, cortical folding or any combination of them). To overcome this limitation, we used Free-Surfer to study how variability in SBM measures was related to the FFM in n = 507 participants from the Human Connectome Project.Neuroticism was associated with thicker cortex and smaller area and folding in prefrontal-temporal regions. Extraversion was linked to thicker pre-cuneus and smaller superior temporal cortex area. Openness was linked to thinner cortex and greater area and folding in prefrontal-parietal regions. Agreeableness was correlated to thinner prefrontal cortex and smaller fusiform gyrus area. Conscientiousness was associated with thicker cortex and smaller area and folding in prefrontal regions. These findings demonstrate that anatomical variability in prefrontal cortices is linked to individual differences in the socio-cognitive dispositions described by the FFM. Cortical thickness and surface area/folding were inversely related each others as a function of different FFM traits (neuroticism, extraversion and consciousness vs openness), which may reflect brain maturational effects that predispose or protect against psychiatric disorders. © The Author (2017). Published by Oxford University Press.

  10. Production of Biodiesel Using Immobilized Lipase and the Characterization of Different Co-Immobilizing Agents and Immobilization Methods

    Directory of Open Access Journals (Sweden)

    Kang Zhao

    2016-08-01

    Full Text Available Lipase from Candida sp. 99–125 is widely employed to catalyzed transesterification and can be used for biodiesel production. In this study, the lipase was immobilized by combined adsorption and entrapment to catalyze biodiesel production from waste cooking oil (WCO via transesterification, and investigating co-immobilizing agents as additives according to the enzyme activity. The addition of the mixed co-immobilizing agents has positive effects on the activities of the immobilized lipase. Three different immobilizing methods were compared by the conversion ratio of biodiesel and structured by Atom Force Microscopy (AFM and Scanning Electron Microscopy (SEM, respectively. It was found that entrapment followed by adsorption was the best method. The effect of the co-immobilizing agent amount, lipase dosage, water content, and reuse ability of the immobilized lipase was investigated. By comparison with previous research, this immobilized lipase showed good reuse ability: the conversion ratio excesses 70% after 10 subsequent reactions, in particular, was better than Novozym435 and TLIM on waste cooking oil for one unit of lipase.

  11. EPDIM peptide-immobilized porous chitosan beads for enhanced wound healing: Preparation, characterizations and in vitro evaluation

    International Nuclear Information System (INIS)

    Bae, Jin Woo; Lee, Joon Hye; Choi, Won Sup; Lee, Dong Sin; Bae, Eun Hee; Park, Ki Dong

    2009-01-01

    EPDIM peptide is known to regulate cellular activities by interacting with α 3 β 1 integrin, which can be contributed to wound healing process. In this study, EPDIM was immobilized onto three-dimensional porous chitosan beads (χtopore) as a scaffold for enhanced wound healing. The significant decrease in contact angle indicates that EPDIM immobilization could lead to the enhanced surface wettability after its immobilization. The immobilized EPDIM was fairly distributed along its surface and the morphology was maintained even after the reaction. The immobilized amount of EPDIM was found to be about 5.68 nmol/mg of χtopore by amino acid analysis. To verify the complete removal of coupling agents after EPDIM immobilization, each coupling agent was quantitatively analyzed by LC-MS. In vitro proliferation rates of both NIH 3T3 and HaCaT showed that EPDIM immobilization onto χtopore could significantly enhance the growth rate of both cells, while the unmodified χtopore did not increase in cell number even after 15 days of culture. Therefore, these results demonstrate that EPDIM peptide-immobilized χtopore can be utilized as an attractive scaffold for enhanced wound healing.

  12. Factors Controlling Summertime Surface Ozone In The Western U.S

    Science.gov (United States)

    Gao, Mei

    In this dissertation we investigate different factors controlling summertime surface ozone (O3) in the western U.S., including the impacts from increased wildfire emissions, the modulation by North American summer monsoon as well as long-range transport of O3 and its precursors from outside of North America. We first analyze the surface ozone observations from the Clean Air Status and Trend Network (CASTNet) using a global chemical transport model (GEOS-Chem) to investigate the impact of biomass burning on surface O3 in the western U.S. (WUS) mountain ranges during the June--October fire season of 2007, one of the stronger fire years in the WUS in the past decade. GEOS-Chem O3 captures the observed seasonal, synoptic and daily variations. Model daily afternoon average surface O3 concentrations at the CASTNet sites are within 2 ppb of the observations, with correlation coefficients of 0.51--0.83 and Taylor scores of 0.64--0.92. Observed maximum daily 8-hour (MAD8) surface O3 concentrations are 37--8 ppb at the sites, while the corresponding model results are higher by 6 ppb on average. Model results show July--September maximum surface O3 enhancement of ~9 ppb on average because of biomass burning. Peaks in fire-contributed surface O3 correspond broadly with high levels of potassium (K), reaffirming a strong fire influence. We find a policy relevant background (PRB) O3 of 45.6 ppb on average during July--September. Fire-contributed O3 accounts for up to 30% of the PRB O3, highest in the intense fire region (Montana, Idaho, and Wyoming) with maxima in August and September. We also examine an unexpected summertime surface O3 minimum (~30--5 ppb) in July--August 2007 observed throughout the Southwestern U.S. (SWUS) by interpreting observations of O3 and rainfall from the Clean Air Status and Trends Network (CASTNet) with a global chemical transport model. The O3 minimum reflects competing chemical and dynamic factors as well as anthropogenic and natural influences. Its

  13. Modification of natural feldspar as support for enzyme immobilization

    International Nuclear Information System (INIS)

    Mohd Basyaruddin Abdul Rahman; Uswatun Hasanah Zaidan; Mahiran Basri; Raja Noor Zaliha Raja Abdul Rahman; Abu Bakar Salleh

    2009-01-01

    The land of Tanah Putih, Gua Musang, Kelantan (Malaysia) is well-known for its wealth in industrial mineral resources, especially aluminosilicate of feldspar and mica. Natural feldspar and mica were physicochemically characterized with regard to X-ray diffraction (XRD), nitrogen sorption analysis and transmission electron microscopy (TEM) techniques for qualitative and quantitative identification of feldspar and mica. They show a good crystallinity, high surface area and uniformity of meso porous structures. For the purpose of this experiment, the aluminosilicate of feldspar was modified either by acid treatment, or grafting the silanol groups present with various functional groups including aminopropyl-, octyl-, vinyl-, mercapto- and glycidoxy-triethoxy silanes, or activation of pre-treated support with glutaraldehyde. These support derivatives were used for further utilization in the immobilization of lipase from Candida rugosa and resulted in various interaction mechanisms between enzyme and introduced supports. It seemed that the features of the functionalized feldspar surfaces provide a preferable environmental host to enable the adsorption of lipase via interfacial adsorption method. Lipase immobilization onto feldspar support were further confirmed by scanning electron microscopy (SEM) coupled with energy dispersive X-ray microanalysis (EDX), transmission electron microscopy (TEM) and infra-red spectroscopy (FTIR) techniques. Enhancement of protein loading (up to 8.22 mg protein/ g support) and immobilization yield (up to 78 %) were shown by modified feldspar-lipase derivatives compared to unmodified feldspar support.(author)

  14. Hyaluronan Immobilized Polyurethane as a Blood Contacting Material

    Directory of Open Access Journals (Sweden)

    Feirong Gong

    2010-01-01

    Full Text Available Hyaluronan (hyaluronic acid, HA was immobilized onto the surface of amino-functionalized polyurethane films with the goal of obtaining a novel kind of biomaterial which had the potential in blood-contacting applications. The amino-functionalized polyurethane was prepared by synthesized acidic polyurethane whose pendant carboxyl groups were treated with an excess amount of 1,3-diaminopropane in the presence of N,N-carbonyldiimidazole (CDI. Attenuated total reflection Fourier transform infrared spectroscopy (ATR-FTIR, Raman spectroscopy (RS, scanning electron microscopy (SEM, and water contact angle measurement were used to confirm the surface changes at each step of treatment, both in morphologies and chemical compositions. APTT and PT results showed that HA immobilization could prolong the blood coagulation time, thus HA-immobilized polyurethane (PU-HA exhibited improved blood compatibility. Cytotoxicity analysis showed that the PU-HA films synthesized in this study were cytocompatible and could support human vein endothelial cells (HUVECs adhesion and proliferation.

  15. Conformal radiotherapy for prostate cancer: contribution of pelvic immobilization and new fiducial markers

    International Nuclear Information System (INIS)

    Garcia, R.; Oozeer, R.; Le Thanh, H.; Chauvet, B.; Toy, B.J.; Reboul, F.

    1997-01-01

    To reduce errors in the positioning of patients treated with external conformal radiotherapy for prostate cancer, we evaluated both the use of an immobilization device and new fiducial markers. The immobilization device consisted of an individual mold made of polyurethane foam. Two sets of skin markers located on the anterior tibial surfaces were used to identify the pelvic isocenter. The patient's position was evaluated by orthogonal port film which were then compared with the original simulation film. Results are presented with respect to orthogonal axes. Comparison with classic procedures without immobilization showed that use of the mold and new fiducial markers led to a decrease in set-up errors which were less than 5 mm. With the use of an immobilization device and optimized techniques for patients' positioning, conformal radiotherapy of prostate cancer is more accurate. (authors)

  16. Reaching (sub-)micrometer resolution of photo-immobilized proteins using diffracted light beams

    DEFF Research Database (Denmark)

    Skovsen, Esben; Neves Petersen, Teresa; Petersen, Steffen B.

    2008-01-01

    , with dimensions as small as a few micrometers. The ultimate size of the immobilized spots is dependent on the focal area of the UV beam. The technology involves light induced formation of free, reactive thiol groups in molecules containing aromatic residues nearby disulphide bridges. It is not only limited...... to immobilizing molecules according to conventional patterns like microarrays, as any bitmap motif can virtually be used a template for patterning. We now show that molecules (proteins) can be immobilized on a surface with any arbitrary pattern according to diffraction patterns of light. The pattern of photo......-immobilized proteins reproduces the diffraction pattern of light expected with the optical setup. Immobilising biomolecules according to diffraction patterns of light will allow achievement of smaller patterns with higher resolution. The flexibility of this new technology leads to any patterns of photo...

  17. Oriented antibody immobilization on self-assembled monolayers applied as impedance biosensors

    Science.gov (United States)

    Tsugimura, Kaiki; Ohnuki, Hitoshi; Wu, Haiyun; Endo, Hideaki; Tsuya, Daiju; Izumi, Mitsuru

    2017-11-01

    Oriented immobilization of antibodies on a sensor chip is crucial for enhancing both the sensitivity and antigen-binding capacity of immunosensors. Here, we report a comparative study of the effect of oriented and random antibody immobilization on the binding efficiency by electrochemical impedance spectroscopy (EIS). Oriented immobilization of anti-myoglobin immunoglobulin G (anti-Myo IgG) was achieved by bonding to an Fc receptor of protein G (PrG) on a self-assembled monolayer (SAM), which results in the myoglobin (Myo) binding sites being exposed outside the sensing surface. Random immobilization of anti-Myo IgG was achieved by direct covalent attachment to the SAM surface. Both immobilizations were applied to interdigitated electrodes to enhance the electrochemical signal, and the Myo biosensor performance was then evaluated by a series of EIS measurements. We found that (i) the rate of the normalized charge transfer resistance for the oriented sample was 3 times higher than that for the random sample and (ii) the detection limit was 0.001 ng/mL, which is the lowest recorded detection limit among Myo immunosensors based on EIS. These findings indicate that oriented antibody immobilization is crucial for preparing highly sensitive EIS-based biosensors.

  18. Evaluation of protein immobilization capacity on various carbon nanotube embedded hydrogel biomaterials

    Energy Technology Data Exchange (ETDEWEB)

    Derkus, Burak, E-mail: burakderkus@gmail.com; Emregul, Kaan Cebesoy; Emregul, Emel

    2015-11-01

    This study investigates effective immobilization of proteins, an important procedure in many fields of bioengineering and medicine, using various biomaterials. Gelatin, alginate and chitosan were chosen as polymeric carriers, and applied in both their composites and nanocomposite forms in combination with carbon nanotubes (CNTs). The prepared nano/composite structures were characterized using scanning electron microscopy (SEM), Fourier-transform infrared spectroscopy (FTIR), thermal gravimetric analysis (TG) and contact angle analysis (CA). Electrochemical impedance spectroscopy analysis revealed gelatin composites in general to exhibit better immobilization performance relative to the native gelatin which can be attributed to enhanced film morphologies of the composite structures. Moreover, superior immobilization efficiencies were obtained with the addition of carbon nanotubes, due to their conducting and surface enhancement features, especially in the gelatin–chitosan structures due to the presence of structural active groups. - Highlights: • Various nanocomposite biomaterials were developed for efficient immobilization of proteins. • CNTs enhance the immobilization efficiency owing to their conducting and surface enhancement features. • Gelatin–chitosan–CNTs structure is promising immobilization matrix thanks to its effective CNTs binding capacity.