Development and characterization of superparamagnetic coatings

OpenAIRE

Kuschnerus I.; Lüdtke-Buzug K.

2015-01-01

Since 2005, Magnetic Particle Imaging (MPI) is handled as a key technology with great potential in medical applications as an imaging method [1]. The superparamagnetic iron oxide nanoparticles (SPIONs) which are already used as a tracer in MPI, combined with various polymers, are being investigated in order to enhance this potential. A combination of polymers such as polyethylene (PE) and polyurethane (PU) and SPIONs could be used as a coating for medical devices, or added to semi-rigid polyu...

PRELIMINARY STUDY ON RETRO-REFLECTIVE COATED PAPER BASED ON MICRO-GLASS BEADS

Institute of Scientific and Technical Information of China (English)

YulongWang; ChuanshanZhao; TaoZhang

2004-01-01

High-reflectivity micro-glass bead, as a kind ofretro-reflective material, is widely used in reflectivefabric or film and other reflective coatings. But it israrely used in coated paper. The retro-reflectivetheory of micro-bead is described. Also the effect ofsize of micro-bead, dosage of binder and differentcolor layers on reflective properties of coated paperare discussed in this article. The results show that itsretro-reflective efficiency is good, equally toreflective fabric or film when the micro-glass bead isused in coated paper.

PRELIMINARY STUDY ON RETRO-REFLECTIVE COATED PAPER BASED ON MICRO-GLASS BEADS

Institute of Scientific and Technical Information of China (English)

Yulong Wang; Chuanshan Zhao; Tao Zhang

2004-01-01

High-reflectivity micro-glass bead, as a kind of retro-reflective material, is widely used in reflective fabric or film and other reflective coatings. But it is rarely used in coated paper. The retro-reflective theory of micro-bead is described. Also the effect of size of micro-bead, dosage of binder and different color layers on reflective properties of coated paper are discussed in this article. The results show that its retro-reflective efficiency is good, equally to reflective fabric or film when the micro-glass bead is used in coated paper.

New Detection Modality for Label-Free Quantification of DNA in Biological Samples via Superparamagnetic Bead Aggregation

Science.gov (United States)

Leslie, Daniel C.; Li, Jingyi; Strachan, Briony C.; Begley, Matthew R.; Finkler, David; Bazydlo, Lindsay L.; Barker, N. Scott; Haverstick, Doris; Utz, Marcel; Landers, James P.

2012-01-01

Combining DNA and superparamagnetic beads in a rotating magnetic field produces multiparticle aggregates that are visually striking, and enables label-free optical detection and quantification of DNA at levels in the picogram per microliter range. DNA in biological samples can be quantified directly by simple analysis of optical images of microfluidic wells placed on a magnetic stirrer without DNA purification. Aggregation results from DNA/bead interactions driven either by the presence of a chaotrope (a nonspecific trigger for aggregation) or by hybridization with oligonucleotides on functionalized beads (sequence-specific). This paper demonstrates quantification of DNA with sensitivity comparable to that of the best currently available fluorometric assays. The robustness and sensitivity of the method enable a wide range of applications, illustrated here by counting eukaryotic cells. Using widely available and inexpensive benchtop hardware, the approach provides a highly accessible low-tech microscale alternative to more expensive DNA detection and cell counting techniques. PMID:22423674

Lab-on-a-Chip Magneto-Immunoassays: How to Ensure Contact between Superparamagnetic Beads and the Sensor Surface

Directory of Open Access Journals (Sweden)

Andreas Hütten

2013-09-01

Full Text Available Lab-on-a-chip immuno assays utilizing superparamagnetic beads as labels suffer from the fact that the majority of beads pass the sensing area without contacting the sensor surface. Different solutions, employing magnetic forces, ultrasonic standing waves, or hydrodynamic effects have been found over the past decades. The first category uses magnetic forces, created by on-chip conducting lines to attract beads towards the sensor surface. Modifications of the magnetic landscape allow for additional transport and separation of different bead species. The hydrodynamic approach uses changes in the channel geometry to enhance the capture volume. In acoustofluidics, ultrasonic standing waves force µm-sized particles onto a surface through radiation forces. As these approaches have their disadvantages, a new sensor concept that circumvents these problems is suggested. This concept is based on the granular giant magnetoresistance (GMR effect that can be found in gels containing magnetic nanoparticles. The proposed design could be realized in the shape of paper-based test strips printed with gel-based GMR sensors.

Influence of Immobilized Biomolecules on Magnetic Bead Plug Formation and Retention in Capillary Electrophoresis

Science.gov (United States)

Henken, Rachel L.; Chantiwas, Rattikan; Gilman, S. Douglass

2012-01-01

Significant changes in the formation and retention of magnetic bead plugs in a capillary during electrophoresis were studied, and it was demonstrated that these effects were due to the type of biological molecule immobilized on the surface of these beads. Three biological molecules, an antibody, an oligonucleotide and alkaline phosphatase, were attached to otherwise identical streptavidin-coated magnetic beads through biotin-avidin binding in order to isolate differences in bead immobilization in a magnetic field resulting from the type of biological molecule immobilized on the bead surface. Alkaline phosphatase also was attached to the magnetic beads using epoxy groups on the bead surfaces (instead of avidin-biotin binding) to study the impact of immobilization chemistry. The formation and retention of magnetic bead plugs were studied quantitatively using light scattering detection of magnetic particles eluting from the bead plugs and qualitatively using microscopy. Both the type of biomolecule immobilized on the magnetic bead surface and the chemistry used to link the biomolecule to the magnetic bead impacted the formation and retention of the bead plugs. PMID:22437880

In vivo biotinylation of recombinant beta-glucosidase enables simultaneous purification and immobilization on streptavidin coated magnetic particles

DEFF Research Database (Denmark)

Alftrén, Johan; Ottow, Kim Ekelund; Hobley, Timothy John

2013-01-01

Beta-glucosidase from Bacillus licheniformis was in vivo biotinylated in Escherichia coli and subsequently immobilized directly from cell lysate on streptavidin coated magnetic particles. In vivo biotinylation was mediated by fusing the Biotin Acceptor Peptide to the C-terminal of beta......-glucosidase and co-expressing the BirA biotin ligase. The approach enabled simultaneous purification and immobilization of the enzyme from crude cell lysate on magnetic particles because of the high affinity and strong interaction between biotin and streptavidin. After immobilization of the biotinylated beta...

Synthesis of streptavidin-conjugated magnetic nanoparticles for DNA detection

International Nuclear Information System (INIS)

Gong Peijun; Peng Zheyang; Wang Yao; Qiao Ru; Mao Weixing; Qian Haisheng; Zhang Mengya; Li Congcong; Shi Shenyuan

2013-01-01

In this paper, we report a fabrication of streptavidin-coated magnetic nanoparticles used for DNA detection. Initially, amino-functionalized Fe 3 O 4 nanoparticles with high saturation magnetization are prepared by a photopolymerization method using allylamine as monomer. It is followed by covalent immobilization of streptavidin onto the particle surface via a two-step reaction using glutaraldehyde as coupling agent. Streptavidin-coated magnetic nanoparticles are characterized and further tested for their ability to capture DNA target after binding biotinylated oligonucleotide probes. The results show that the products (∼27.2 nm) have a maximum biotin-binding capacity of 0.71 nmol mg −1 when the immobilization reaction is conducted with a mass ratio of streptavidin to magnetic carriers above 0.2 in phosphate buffered saline (pH 7.4) for 24 h. In addition, highly negative ζ-potential and good magnetic susceptibility of the nanocomposites make them applicable for DNA collection and detection, which is verified by the results from the preliminary application of streptavidin-coated magnetic nanoparticles in DNA detection. Therefore, the magnetic nanoparticles provide a promising approach for rapid collection and detection of gene.

Observation of the dynamics of magnetically induced chains of sub-micron superparamagnetic beads in aqueous solutions by laser light scattering

International Nuclear Information System (INIS)

Tanizawa, Y; Tashiro, T; Sandhu, A; Ko, P J

2013-01-01

Optical monitoring the behaviour of magnetically induced self-assembled chains of superparamagnetic beads (SPBs) are of interest for biomedical applications such as biosensors. However, it is difficult to directly monitor magnetically induced self-assembly of sub-micron nano-beads with conventional optical microscopes. Here, we describe the optical observation of the dynamics of magnetically induced self-assembled rotating chains of 130 nm SPBs in aqueous solutions by laser light scattering. Magnetic fields of ∼1 kOe were applied to control the self-assembly chains of SPBs and their behaviour analyzed by monitoring the intensity of laser light scattered from the chain structures. We compared the light scattering from chains that were formed only by the application of external fields with chains formed by beads functionalized by EDC, where chemical reactions lead to the bonding of individual beads to form chains. The EDC experiments are a precursor to experiments on molecular recognition applications for biomedical diagnostics.

Preparation of acetaminophen capsules containing beads prepared by hot-melt direct blend coating.

Science.gov (United States)

Pham, Loan; Christensen, John M

2014-02-01

Twelve hydrophobic coating agents were assessed for their effects on drug release after coating sugar cores by a flexible hot-melt coating method using direct blending. Drug-containing pellets were also produced and used as cores. The cores were coated with single or double wax layers containing acetaminophen (APAP). The harder the wax, the slower the resultant drug releases from single-coated beads. Wax coating can be deposited on cores up to 28% of the beads final weight and reaching 58% with wax and drug. Carnauba-coated beads dissolved in approximately 6 h releasing 80% of the loaded drug. Applying another wax layer extended drug release over 20 h, while still delivering 80% of the loaded drug. When drug-containing pellets (33-58% drug loading) were used as cores, double wax-coated pellets exhibited a near zero-order drug release for 16 h, releasing 80% of the loaded drug delivering 18 mg/h. The simple process of hot-melt coating by direct blending of pellet-containing drug-coated formulations provides excellent options for immediate and sustained release formulations when higher lipid coating or drug loading is warranted. Predicted plasma drug concentration time profiles using convolution and in vitro drug release properties of the beads were performed for optimal formulations.

Peptide nucleic acid probe for protein affinity purification based on biotin-streptavidin interaction and peptide nucleic acid strand hybridization.

Science.gov (United States)

Tse, Jenny; Wang, Yuanyuan; Zengeya, Thomas; Rozners, Eriks; Tan-Wilson, Anna

2015-02-01

We describe a new method for protein affinity purification that capitalizes on the high affinity of streptavidin for biotin but does not require dissociation of the biotin-streptavidin complex for protein retrieval. Conventional reagents place both the selectively reacting group (the "warhead") and the biotin on the same molecule. We place the warhead and the biotin on separate molecules, each linked to a short strand of peptide nucleic acid (PNA), synthetic polymers that use the same bases as DNA but attached to a backbone that is resistant to attack by proteases and nucleases. As in DNA, PNA strands with complementary base sequences hybridize. In conditions that favor PNA duplex formation, the warhead strand (carrying the tagged protein) and the biotin strand form a complex that is held onto immobilized streptavidin. As in DNA, the PNA duplex dissociates at moderately elevated temperature; therefore, retrieval of the tagged protein is accomplished by a brief exposure to heat. Using iodoacetate as the warhead, 8-base PNA strands, biotin, and streptavidin-coated magnetic beads, we demonstrate retrieval of the cysteine protease papain. We were also able to use our iodoacetyl-PNA:PNA-biotin probe for retrieval and identification of a thiol reductase and a glutathione transferase from soybean seedling cotyledons. Copyright © 2014 Elsevier Inc. All rights reserved.

Evaluation of Antigen-Conjugated Fluorescent Beads to Identify Antigen-Specific B Cells

Directory of Open Access Journals (Sweden)

Isabel Correa

2018-03-01

Full Text Available Selection of single antigen-specific B cells to identify their expressed antibodies is of considerable interest for evaluating human immune responses. Here, we present a method to identify single antibody-expressing cells using antigen-conjugated fluorescent beads. To establish this, we selected Folate Receptor alpha (FRα as a model antigen and a mouse B cell line, expressing both the soluble and the membrane-bound forms of a human/mouse chimeric antibody (MOv18 IgG1 specific for FRα, as test antibody-expressing cells. Beads were conjugated to FRα using streptavidin/avidin-biotin bridges and used to select single cells expressing the membrane-bound form of anti-FRα. Bead-bound cells were single cell-sorted and processed for single cell RNA retrotranscription and PCR to isolate antibody heavy and light chain variable regions. Variable regions were then cloned and expressed as human IgG1/k antibodies. Like the original clone, engineered antibodies from single cells recognized native FRα. To evaluate whether antigen-coated beads could identify specific antibody-expressing cells in mixed immune cell populations, human peripheral blood mononuclear cells (PBMCs were spiked with test antibody-expressing cells. Antigen-specific cells could comprise up to 75% of cells selected with antigen-conjugated beads when the frequency of the antigen-positive cells was 1:100 or higher. In PBMC pools, beads conjugated to recombinant antigens FRα and HER2 bound antigen-specific anti-FRα MOv18 and anti-HER2 Trastuzumab antibody-expressing cells, respectively. From melanoma patient-derived B cells selected with melanoma cell line-derived protein-coated fluorescent beads, we generated a monoclonal antibody that recognized melanoma antigen-coated beads. This approach may be further developed to facilitate analysis of B cells and their antibody profiles at the single cell level and to help unravel humoral immune repertoires.

Evaluation of Antigen-Conjugated Fluorescent Beads to Identify Antigen-Specific B Cells.

Science.gov (United States)

Correa, Isabel; Ilieva, Kristina M; Crescioli, Silvia; Lombardi, Sara; Figini, Mariangela; Cheung, Anthony; Spicer, James F; Tutt, Andrew N J; Nestle, Frank O; Karagiannis, Panagiotis; Lacy, Katie E; Karagiannis, Sophia N

2018-01-01

Selection of single antigen-specific B cells to identify their expressed antibodies is of considerable interest for evaluating human immune responses. Here, we present a method to identify single antibody-expressing cells using antigen-conjugated fluorescent beads. To establish this, we selected Folate Receptor alpha (FRα) as a model antigen and a mouse B cell line, expressing both the soluble and the membrane-bound forms of a human/mouse chimeric antibody (MOv18 IgG1) specific for FRα, as test antibody-expressing cells. Beads were conjugated to FRα using streptavidin/avidin-biotin bridges and used to select single cells expressing the membrane-bound form of anti-FRα. Bead-bound cells were single cell-sorted and processed for single cell RNA retrotranscription and PCR to isolate antibody heavy and light chain variable regions. Variable regions were then cloned and expressed as human IgG1/k antibodies. Like the original clone, engineered antibodies from single cells recognized native FRα. To evaluate whether antigen-coated beads could identify specific antibody-expressing cells in mixed immune cell populations, human peripheral blood mononuclear cells (PBMCs) were spiked with test antibody-expressing cells. Antigen-specific cells could comprise up to 75% of cells selected with antigen-conjugated beads when the frequency of the antigen-positive cells was 1:100 or higher. In PBMC pools, beads conjugated to recombinant antigens FRα and HER2 bound antigen-specific anti-FRα MOv18 and anti-HER2 Trastuzumab antibody-expressing cells, respectively. From melanoma patient-derived B cells selected with melanoma cell line-derived protein-coated fluorescent beads, we generated a monoclonal antibody that recognized melanoma antigen-coated beads. This approach may be further developed to facilitate analysis of B cells and their antibody profiles at the single cell level and to help unravel humoral immune repertoires.

Evaluation of Antigen-Conjugated Fluorescent Beads to Identify Antigen-Specific B Cells

Science.gov (United States)

Correa, Isabel; Ilieva, Kristina M.; Crescioli, Silvia; Lombardi, Sara; Figini, Mariangela; Cheung, Anthony; Spicer, James F.; Tutt, Andrew N. J.; Nestle, Frank O.; Karagiannis, Panagiotis; Lacy, Katie E.; Karagiannis, Sophia N.

2018-01-01

Selection of single antigen-specific B cells to identify their expressed antibodies is of considerable interest for evaluating human immune responses. Here, we present a method to identify single antibody-expressing cells using antigen-conjugated fluorescent beads. To establish this, we selected Folate Receptor alpha (FRα) as a model antigen and a mouse B cell line, expressing both the soluble and the membrane-bound forms of a human/mouse chimeric antibody (MOv18 IgG1) specific for FRα, as test antibody-expressing cells. Beads were conjugated to FRα using streptavidin/avidin-biotin bridges and used to select single cells expressing the membrane-bound form of anti-FRα. Bead-bound cells were single cell-sorted and processed for single cell RNA retrotranscription and PCR to isolate antibody heavy and light chain variable regions. Variable regions were then cloned and expressed as human IgG1/k antibodies. Like the original clone, engineered antibodies from single cells recognized native FRα. To evaluate whether antigen-coated beads could identify specific antibody-expressing cells in mixed immune cell populations, human peripheral blood mononuclear cells (PBMCs) were spiked with test antibody-expressing cells. Antigen-specific cells could comprise up to 75% of cells selected with antigen-conjugated beads when the frequency of the antigen-positive cells was 1:100 or higher. In PBMC pools, beads conjugated to recombinant antigens FRα and HER2 bound antigen-specific anti-FRα MOv18 and anti-HER2 Trastuzumab antibody-expressing cells, respectively. From melanoma patient-derived B cells selected with melanoma cell line-derived protein-coated fluorescent beads, we generated a monoclonal antibody that recognized melanoma antigen-coated beads. This approach may be further developed to facilitate analysis of B cells and their antibody profiles at the single cell level and to help unravel humoral immune repertoires. PMID:29628923

A polyamidoamine dendrimer-streptavidin supramolecular architecture for biosensor development.

Science.gov (United States)

Soda, N; Arotiba, O A

2017-12-01

A novel polyamidoamine dendrimer-streptavidin supramolecular architecture suitable as a versatile platform for biosensor development is reported. The dendrimer was electrodeposited on a glassy carbon electrode via cyclic voltammetry. The dendrimer electrode was further modified with streptavidin by electrostatic attraction upon drop coating. The platform i.e. the dendrimer-streptavidin modified electrode was electrochemically interrogated in phosphate buffer, ferrocyanide and H 2 O 2 . The dendrimer-streptavidin platform was used in the preparation of a simple DNA biosensor as a proof of concept. The supramolecular architecture of dendrimer-streptavidin was stable, electroactive and thus lends itself as a versatile immobilisation layer for any biotinylated bioreceptors in biosensor development. Copyright © 2017 Elsevier B.V. All rights reserved.

Cultivation of human dermal fibroblasts and epidermal keratinocytes on keratin-coated silica bead substrates.

Science.gov (United States)

Tan, Bee Yi; Nguyen, Luong T H; Kim, Hyo-Sop; Kim, Jae-Ho; Ng, Kee Woei

2017-10-01

Human hair keratin is promising as a bioactive material platform for various biomedical applications. To explore its versatility further, human hair keratin was coated onto monolayers of silica beads to produce film-like substrates. This combination was hypothesized to provide a synergistic effect in improving the biochemical properties of the resultant composite. Atomic force microscopy analysis showed uniform coatings of keratin on the silica beads with a slight increase in the resulting surface roughness. Keratin-coated silica beads had higher surface energy and relatively lower negative charge than those of bare silica beads. To investigate cell response, human dermal fibroblasts (HDFs), and human epidermal keratinocytes (HEKs) were cultured on the substrates over 4 days. Results showed that keratin coatings significantly enhanced the metabolic activity of HDFs and encouraged cell spreading but did not exert any significant effects on HEKs. HDF expression of collagen I was significantly more intense on the keratin-coated compared to the bare silica substrates. Furthermore, HDF secretion of various cytokines suggested that keratin coatings triggered active cell responses related to wound healing. Collectively, our study demonstrated that human hair keratin-coated silica bead monolayers have the potential to modulate HDF behavior in culture and may be exploited further. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 2789-2798, 2017. © 2017 Wiley Periodicals, Inc.

Highly Specific Binding on Antifouling Zwitterionic Polymer-Coated Microbeads as Measured by Flow Cytometry.

Science.gov (United States)

van Andel, Esther; de Bus, Ian; Tijhaar, Edwin J; Smulders, Maarten M J; Savelkoul, Huub F J; Zuilhof, Han

2017-11-08

Micron- and nano-sized particles are extensively used in various biomedical applications. However, their performance is often drastically hampered by the nonspecific adsorption of biomolecules, a process called biofouling, which can cause false-positive and false-negative outcomes in diagnostic tests. Although antifouling coatings have been extensively studied on flat surfaces, their use on micro- and nanoparticles remains largely unexplored, despite the widespread experimental (specifically, clinical) uncertainties that arise because of biofouling. Here, we describe the preparation of magnetic micron-sized beads coated with zwitterionic sulfobetaine polymer brushes that display strong antifouling characteristics. These coated beads can then be equipped with recognition elements of choice, to enable the specific binding of target molecules. First, we present a proof of principle with biotin-functionalized beads that are able to specifically bind fluorescently labeled streptavidin from a complex mixture of serum proteins. Moreover, we show the versatility of the method by demonstrating that it is also possible to functionalize the beads with mannose moieties to specifically bind the carbohydrate-binding protein concanavalin A. Flow cytometry was used to show that thus-modified beads only bind specifically targeted proteins, with minimal/near-zero nonspecific protein adsorption from other proteins that are present. These antifouling zwitterionic polymer-coated beads, therefore, provide a significant advancement for the many bead-based diagnostic and other biosensing applications that require stringent antifouling conditions.

Dextran-coated superparamagnetic amorphous Fe–Co nanoalloy for magnetic resonance imaging applications

International Nuclear Information System (INIS)

An, Lu; Yu, Yanrong; Li, Xuejian; Liu, Wei; Yang, Hong; Wu, Dongmei; Yang, Shiping

2014-01-01

Graphical abstract: A dextran-coated Fe–Co nanoalloy was developed serving as a sensitive contrast agent for magnetic resonance imaging applications. - Highlights: • Amorphous Fe–Co nanoalloy was prepared via wet chemical reduction approach. • The Fe–Co nanoalloy is water-soluble, stable, and biocompatible. • The Fe–Co nanoalloy is superparamagnetic. • The Fe–Co nanoalloy exhibits T 2 -weighted MR enhancement both in vitro and in vivo. - Abstract: For magnetic resonance imaging applications, a facile approach for water-soluble dextran coated amorphous Fe–Co nanoalloy was developed. The as-synthesized nanoalloy had a diameter of 9 nm with a narrow size distribution and showed superparamagnetic property with a saturated magnetization (Ms) of 25 emu/g. In vitro cytotoxicity test revealed that it was biocompatible at a concentration below 120 μg/mL. It can be uptaken by HeLa cells effectively and resulted in the obvious T 2 effect after internalization. Biodistribution studies in conjunction with inductively coupled plasma-atomic emission spectroscopy (ICP-AES) confirmed that Fe–Co nanoalloy was preferentially accumulated in lung and spleen after intravenous injection for 4 h. In vivo MRI, dextran-coated Fe–Co nanoalloy can serve as a sensitive contrast agent for MR imaging, especially in the spleen, so we believe that it maybe hold great promise for diagnosis of splenic disease by appropriately functionalizing their surface

Dextran-coated superparamagnetic amorphous Fe–Co nanoalloy for magnetic resonance imaging applications

Energy Technology Data Exchange (ETDEWEB)

An, Lu; Yu, Yanrong; Li, Xuejian; Liu, Wei [The Key Laboratory of Resource Chemistry of Ministry of Education and Shanghai Key Laboratory of Rare Earth Functional Materials, Department of Chemistry, Shanghai Normal University, Shanghai 200234 (China); Yang, Hong, E-mail: yanghong@shnu.edu.cn [The Key Laboratory of Resource Chemistry of Ministry of Education and Shanghai Key Laboratory of Rare Earth Functional Materials, Department of Chemistry, Shanghai Normal University, Shanghai 200234 (China); Wu, Dongmei [Shanghai Key Laboratory of Magnetic Resonance, Department of Physics, East China Normal University, 3663 North Zhongshan Road, Shanghai 200062 (China); Yang, Shiping, E-mail: shipingy@shnu.edu.cn [The Key Laboratory of Resource Chemistry of Ministry of Education and Shanghai Key Laboratory of Rare Earth Functional Materials, Department of Chemistry, Shanghai Normal University, Shanghai 200234 (China)

2014-01-01

Graphical abstract: A dextran-coated Fe–Co nanoalloy was developed serving as a sensitive contrast agent for magnetic resonance imaging applications. - Highlights: • Amorphous Fe–Co nanoalloy was prepared via wet chemical reduction approach. • The Fe–Co nanoalloy is water-soluble, stable, and biocompatible. • The Fe–Co nanoalloy is superparamagnetic. • The Fe–Co nanoalloy exhibits T{sub 2}-weighted MR enhancement both in vitro and in vivo. - Abstract: For magnetic resonance imaging applications, a facile approach for water-soluble dextran coated amorphous Fe–Co nanoalloy was developed. The as-synthesized nanoalloy had a diameter of 9 nm with a narrow size distribution and showed superparamagnetic property with a saturated magnetization (Ms) of 25 emu/g. In vitro cytotoxicity test revealed that it was biocompatible at a concentration below 120 μg/mL. It can be uptaken by HeLa cells effectively and resulted in the obvious T{sub 2} effect after internalization. Biodistribution studies in conjunction with inductively coupled plasma-atomic emission spectroscopy (ICP-AES) confirmed that Fe–Co nanoalloy was preferentially accumulated in lung and spleen after intravenous injection for 4 h. In vivo MRI, dextran-coated Fe–Co nanoalloy can serve as a sensitive contrast agent for MR imaging, especially in the spleen, so we believe that it maybe hold great promise for diagnosis of splenic disease by appropriately functionalizing their surface.

Echicetin coated polystyrene beads: a novel tool to investigate GPIb-specific platelet activation and aggregation.

Directory of Open Access Journals (Sweden)

Alexey Navdaev

Full Text Available von Willebrand factor/ristocetin (vWF/R induces GPIb-dependent platelet agglutination and activation of αIIbβ3 integrin, which also binds vWF. These conditions make it difficult to investigate GPIb-specific signaling pathways in washed platelets. Here, we investigated the specific mechanisms of GPIb signaling using echicetin-coated polystyrene beads, which specifically activate GPIb. We compared platelet activation induced by echicetin beads to vWF/R. Human platelets were stimulated with polystyrene beads coated with increasing amounts of echicetin and platelet activation by echicetin beads was then investigated to reveal GPIb specific signaling. Echicetin beads induced αIIbβ3-dependent aggregation of washed platelets, while under the same conditions vWF/R treatment led only to αIIbβ3-independent platelet agglutination. The average distance between the echicetin molecules on the polystyrene beads must be less than 7 nm for full platelet activation, while the total amount of echicetin used for activation is not critical. Echicetin beads induced strong phosphorylation of several proteins including p38, ERK and PKB. Synergistic signaling via P2Y12 and thromboxane receptor through secreted ADP and TxA2, respectively, were important for echicetin bead triggered platelet activation. Activation of PKG by the NO/sGC/cGMP pathway inhibited echicetin bead-induced platelet aggregation. Echicetin-coated beads are powerful and reliable tools to study signaling in human platelets activated solely via GPIb and GPIb-triggered pathways.

Echicetin Coated Polystyrene Beads: A Novel Tool to Investigate GPIb-Specific Platelet Activation and Aggregation

Science.gov (United States)

Petunin, Alexey; Clemetson, Kenneth J.; Gambaryan, Stepan; Walter, Ulrich

2014-01-01

von Willebrand factor/ristocetin (vWF/R) induces GPIb-dependent platelet agglutination and activation of αIIbβ3 integrin, which also binds vWF. These conditions make it difficult to investigate GPIb-specific signaling pathways in washed platelets. Here, we investigated the specific mechanisms of GPIb signaling using echicetin-coated polystyrene beads, which specifically activate GPIb. We compared platelet activation induced by echicetin beads to vWF/R. Human platelets were stimulated with polystyrene beads coated with increasing amounts of echicetin and platelet activation by echicetin beads was then investigated to reveal GPIb specific signaling. Echicetin beads induced αIIbβ3-dependent aggregation of washed platelets, while under the same conditions vWF/R treatment led only to αIIbβ3-independent platelet agglutination. The average distance between the echicetin molecules on the polystyrene beads must be less than 7 nm for full platelet activation, while the total amount of echicetin used for activation is not critical. Echicetin beads induced strong phosphorylation of several proteins including p38, ERK and PKB. Synergistic signaling via P2Y12 and thromboxane receptor through secreted ADP and TxA2, respectively, were important for echicetin bead triggered platelet activation. Activation of PKG by the NO/sGC/cGMP pathway inhibited echicetin bead-induced platelet aggregation. Echicetin-coated beads are powerful and reliable tools to study signaling in human platelets activated solely via GPIb and GPIb-triggered pathways. PMID:24705415

Characterization of Antigen-Specific B Cells Using Nominal Antigen-Coated Flow-Beads

Science.gov (United States)

Akl, Ahmed; Lepetit, Maud; Crochette, Romain; Giral, Magali; Lepourry, Julie; Pallier, Annaick; Castagnet, Stéphanie; Dugast, Emilie; Guillot-Gueguen, Cécile; Jacq-Foucher, Marylène; Saulquin, Xavier; Cesbron, Anne; Laplaud, David; Nicot, Arnaud; Brouard, Sophie; Soulillou, Jean-Paul

2013-01-01

In order to characterize the reactivity of B cells against nominal antigens, a method based on the coupling of antigens onto the surface of fluorescent core polystyrene beads was developed. We first demonstrate that murine B cells with a human MOG-specific BCR are able to interact with MOG-coated beads and do not recognize beads coated with human albumin or pp65. B cells purified from human healthy volunteer blood or immunized individuals were tested for their ability to interact with various nominal antigens, including viral, vaccine, self and alloantigens, chosen for their usefulness in studying a variety of pathological processes. A substantial amount of B cells binding self-antigen MOG-coated beads can be detected in normal blood. Furthermore, greater frequencies of B cell against anti-Tetanic Toxin or anti-EBNA1 were observed in primed individuals. This method can reveal increased frequencies of anti-HLA committed B cells in patients with circulating anti-HLA antibodies compared to unsensitized patients and normal individuals. Of interest, those specific CD19 cells were preferentially identified within CD27−IgD+ (i-e naïve) subset. These observations suggest that a broad range of medical situations could benefit from a tool that allows the detection, the quantification and the characterization of antigen-specific blood B cells. PMID:24386360

Evaluation of superparamagnetic and biocompatible properties of mesoporous silica coated cobalt ferrite nanoparticles synthesized via microwave modified Pechini method

Energy Technology Data Exchange (ETDEWEB)

Gharibshahian, M. [Faculty of New Sciences and Technologies, Semnan University, Semnan (Iran, Islamic Republic of); Mirzaee, O., E-mail: O_mirzaee@semnan.ac.ir [Faculty of Materials and Metallurgical Engineering, Semnan University, Semnan (Iran, Islamic Republic of); Nourbakhsh, M.S. [Faculty of New Sciences and Technologies, Semnan University, Semnan (Iran, Islamic Republic of)

2017-03-01

Cobalt ferrite nano particles were synthesized by Pechini sol-gel method and calcined at 700 °C in electrical and microwave furnace. The microwave calcined sample was coated with mesoporous silica by hydrothermal method. Characterization was performed by XRD, FESEM, TEM, VSM, BET and FTIR analysis. The cytotoxicity was evaluated by MTT assay with 3T3 fibroblast cells. The XRD and FTIR results confirmed spinal formation in both cases and verified the formation of silica coating on the nanoparticles. For microwave calcination, The XRD and SEM results demonstrated smaller and flat adhesion forms of nanoparticles with the average size of 15 nm. The VSM results demonstrated nearly superparamagnetic nanoparticles with significant saturation magnetization equal to 64 emu/g. By coating, saturation magnetization was decreased to 36 emu/g. Moreover, the BET results confirmed the formation of mesoporous coating with the average pore diameters of 2.8 nm and average pore volume of 0.82 cm{sup 3} g{sup −1}. Microwave calcined nanoparticles had the best structural and magnetic properties. - Highlights: • CoFe{sub 2}O{sub 4} nanoparticles were synthesized using the microwave modified Pechini method. • The Effect of calcination route and silica coating on NPs properties was studied. • The nearly superparamagnetic nanoparticles were achieved by microwave calcination. • MFC NPs had the best magnetic properties and MTT assay showed no toxicity for MFC-MSC NPs. • A useful scheme was designed to achieve biological superparamagnetic core/shell NPs.

Zanamivir immobilized magnetic beads for voltammetric measurement of neuraminidase at gold-modified boron doped diamond electrode

Energy Technology Data Exchange (ETDEWEB)

Wahyuni, Wulan Tri, E-mail: wulantriws@gmail.com [Department of Chemistry, Faculty of Mathematics and Natural Sciences, Bogor Agricultural University, Kampus IPB Darmaga, Bogor 16680 (Indonesia); Department of Chemistry, FMIPA, Universitas Indonesia, Kampus UI Depok (Indonesia); Ivandini, Tribidasari A.; Saepudin, Endang [Department of Chemistry, FMIPA, Universitas Indonesia, Kampus UI Depok (Indonesia); Einaga, Yasuaki [Department of Chemistry, Faculty of Science and Technology, Keio University, Hiyoshi 3-14-1, Yokohama 223-8522 (Japan); CREST, JST, 3-14-1 Hiyoshi, Yokohama 223-8522 (Japan)

2016-04-19

Biomolecule modified magnetic beads has been widely used in separation and sensing process. This study used streptavidin modified magnetic beads to immobilize biotin modified zanamivir. Biotin-streptavidin affinity facilitates immobilization of zanamivir on magnetic beads. Then interaction of zanamivir and neuraminidase was adopted as basic for enzyme detection. Detection of neuraminidase was performed at gold modified BDD using cyclic voltammetry technique. The measurement was carried out based on alteration of electrochemical signals of working electrode as neuraminidase response. The result showed that zanamivir was successfully immobilized on magnetic beads. The optimum amount of magnetic beads for zanamivir immobilization was 120 ug. Linear responses of neuraminidase were detected in concentration range of 0-15 mU. Detection limit (LOD) of measurement was 2.32 mU (R2 = 0.959) with precision as % RSD of 1.41%. Measurement of neuraminidase on magnetic beads could be also performed in the presence of mucin matrix. The linearity range was 0-8 mU with LOD of 0.64 mU (R2 = 0.950) and % RSD of 7.25%.

Development of the rectal dosage form with silver-coated glass beads for local-action applications in lower sections of the gastrointestinal tract.

Science.gov (United States)

Siczek, Krzysztof; Fichna, Jakub; Zatorski, Hubert; Karolewicz, Bożena; Klimek, Leszek; Owczarek, Artur

2018-03-01

Recent findings indicating the anti-inflammatory action of silver preparations through modulation of the gut microbiota and apoptosis of inflammatory cells predestine silver use in inflammatory bowel disease (IBD). The aim of our study was to validate the possibility of effective silver release from silver-coated glass beads for anti-inflammatory local application in the lower sections of the gastrointestinal (GI) tract. Silver-coated glass beads were prepared using magnetron method. Release of silver from the silver-coated glass bead surface was carried out in BIO-DIS reciprocating cylinder apparatus. Erosion of silver coating and indirect estimation of the silver release dynamics was assessed using scanning electron microscope. Rectal suppositories containing silver-coated glass beads were prepared using five different methods (M1-M5) and X-ray scanned for their composition. The XR microanalysis and the chemical composition analysis evidenced for a rapid (within 30 min) release of nearly 50% of silver from the coating of the glass beads, which remained stable up to 24 h of incubation. The most homogeneous distribution of beads in the entire volume of the suppository was obtained for formulation M5, where the molten base was poured into mold placed in an ice bath, and the beads were added after 10 s. Our study is the first to present the concept of enclosing silver-coated glass beads in the lipophilic suppository base to attenuate inflammation in the lower GI tract and promises efficient treatment with reduced side effects.

Fibroblast receptor for cell-substratum adhesion: studies on the interaction of baby hamster kidney cells with latex beads coated by cold insoluble globulin (plasma fibronectin)

OpenAIRE

1980-01-01

Studies were carried out on the interactions of uncharged latex beads (0.76 micrometer) with baby hamster kidney cells. Binding of beads to the cells occurred if the beads were coated by cold insoluble globulin (CIG) (plasma fibronectin) but not if the beads were coated by bovine albumin. Bovine albumin-coated beads did not bind to the cells even in the presence of excess CIG in the incubation medium. Binding of beads occurred randomly over the entire surfaces of cells in suspension. However,...

Magnetic and relaxometric properties of polyethylenimine-coated superparamagnetic MRI contrast agents

International Nuclear Information System (INIS)

Corti, M.; Lascialfari, A.; Marinone, M.; Masotti, A.; Micotti, E.; Orsini, F.; Ortaggi, G.; Poletti, G.; Innocenti, C.; Sangregorio, C.

2008-01-01

Novel systems to be employed as superparamagnetic contrast agents (CA) for magnetic resonance imaging (MRI) have been synthesized. These compounds are composed of an iron oxide magnetic core coated by polyethylenimine (PEI) or carboxylated polyethylenimine (PEI-COOH). The aim of the present work was to prepare and study new nanostructured systems (with better or at least comparable relaxivities, R 1 and R 2 , with respect to the commercial ones) with controlled, almost monodisperse average dimensions and shape, as candidates for molecular targeting. By means of atomic force microscopy (AFM) measurements we determined the average diameter, of the order of 200 nm, and the shape of the particles. The superparamagnetic behavior was assessed by SQUID measurements. From X-ray data the estimated average diameters of the magnetic cores were found to be ∼5.8 nm for PEI-COOH60 and ∼20 nm for the compound named PEI25. By NMR-dispersion (NMRD), we found that PEI-COOH60 presents R 1 and R 2 relaxivities slightly lower than Endorem. The experimental results suggest that these novel compounds can be used as MRI CA

Superparamagnetic poly(methyl methacrylate) beads for nattokinase purification from fermentation broth.

Science.gov (United States)

Yang, Chengli; Xing, Jianmin; Guan, Yueping; Liu, Huizhou

2006-09-01

An effective method for purification of nattokinase from fermentation broth using magnetic poly(methyl methacrylate) (PMMA) beads immobilized with p-aminobenzamidine was proposed in this study. Firstly, magnetic PMMA beads with a narrow size distribution were prepared by spraying suspension polymerization. Then, they were highly functionalized via transesterification reaction with polyethylene glycol. The surface hydroxyl-modified magnetic beads obtained were further modified with chloroethylamine to transfer the surface amino-modified magnetic functional beads. The morphology and surface functionality of the magnetic beads were examined by scanning electron microscopy and Fourier transform infrared. An affinity ligand, p-aminobenzamidine was covalently immobilized to the amino-modified magnetic beads by the glutaraldehyde method for nattokinase purification directly from the fermentation broth. The purification factor and the recovery of the enzyme activity were found to be 8.7 and 85%, respectively. The purification of nattokinase from fermentation broth by magnetic beads only took 40 min, which shows a very fast purification of nattokinase compared to traditional purification methods.

Synthesis and characterization of the superparamagnetic iron oxide nanoparticles modified with cationic chitosan and coated with silica shell

Energy Technology Data Exchange (ETDEWEB)

Lewandowska-Łańcucka, Joanna, E-mail: lewandow@chemia.uj.edu.pl [Faculty of Chemistry, Jagiellonian University, Ingardena 3, 30-060 Krakow (Poland); Staszewska, Magdalena; Szuwarzyński, Michał; Kępczyński, Mariusz [Faculty of Chemistry, Jagiellonian University, Ingardena 3, 30-060 Krakow (Poland); Romek, Marek [Department of Cytology and Histology, Institute of Zoology, Jagiellonian University, Gronostajowa 9, 30-387 Krakow (Poland); Tokarz, Waldemar [Department of Solid State Physics, AGH University of Science and Technology, al. A. Mickiewicza 30, 30-059 Krakow (Poland); Szpak, Agnieszka; Kania, Gabriela; Nowakowska, Maria [Faculty of Chemistry, Jagiellonian University, Ingardena 3, 30-060 Krakow (Poland)

2014-02-15

Highlights: • The new, facile methodology for synthesis of silica covered SPIONs is proposed. • The SPIONs was modified with cationic chitosan and coated with silica shell. • Negatively charged, rounded in shape particles of ca. 330 nm were obtained. • The product exhibits the superparamagnetic properties. • The product properties imply its potential applications in biomedicine areas. -- Abstract: Novel method for synthesis of superparamagnetic iron oxide nanoparticles (SPION) modified with a cationic chitosan (CCh) and coated with a silica shell, SPION-CCh-SiO{sub 2} was developed. The process was carried out in two steps. In the first step the chitosan coated SPIONs were obtained by co-precipitation of Fe{sup 2+} and Fe{sup 3+} with ammonium hydroxide in aqueous solution of CCh. In the second one, the silica shell is formed on their surfaces. The formation of SPION-CCh-SiO{sub 2} was achieved by direct decomposition of tetraethoxysilane (TEOS) adsorbed on a surface of SPION-CCh dispersed in aqueous phase under sonication and mechanical stirring at room temperature. The chemical composition and physicochemical properties of the materials were determined using X-ray Diffraction (XRD), Fourier Transform Infrared Spectroscopy (FTIR), Dynamic Light Scattering (DLS) and zeta potential measurements. The morphology of the particles was evaluated by Transmission Electron Microscopy (TEM) and Scanning Electron Microscopy (SEM). Magnetic properties were confirmed using Atomic Force Microscopy/Magnetic Force Microscopy (AFM/MFM) and magnetization measurements. The resulting products are negatively charged, rounded in shape and exhibit the superparamagnetic properties what implies their potential applications in engineering and biomedicine areas.

Synthesis and characterization of the superparamagnetic iron oxide nanoparticles modified with cationic chitosan and coated with silica shell

International Nuclear Information System (INIS)

Lewandowska-Łańcucka, Joanna; Staszewska, Magdalena; Szuwarzyński, Michał; Kępczyński, Mariusz; Romek, Marek; Tokarz, Waldemar; Szpak, Agnieszka; Kania, Gabriela; Nowakowska, Maria

2014-01-01

Highlights: • The new, facile methodology for synthesis of silica covered SPIONs is proposed. • The SPIONs was modified with cationic chitosan and coated with silica shell. • Negatively charged, rounded in shape particles of ca. 330 nm were obtained. • The product exhibits the superparamagnetic properties. • The product properties imply its potential applications in biomedicine areas. -- Abstract: Novel method for synthesis of superparamagnetic iron oxide nanoparticles (SPION) modified with a cationic chitosan (CCh) and coated with a silica shell, SPION-CCh-SiO 2 was developed. The process was carried out in two steps. In the first step the chitosan coated SPIONs were obtained by co-precipitation of Fe 2+ and Fe 3+ with ammonium hydroxide in aqueous solution of CCh. In the second one, the silica shell is formed on their surfaces. The formation of SPION-CCh-SiO 2 was achieved by direct decomposition of tetraethoxysilane (TEOS) adsorbed on a surface of SPION-CCh dispersed in aqueous phase under sonication and mechanical stirring at room temperature. The chemical composition and physicochemical properties of the materials were determined using X-ray Diffraction (XRD), Fourier Transform Infrared Spectroscopy (FTIR), Dynamic Light Scattering (DLS) and zeta potential measurements. The morphology of the particles was evaluated by Transmission Electron Microscopy (TEM) and Scanning Electron Microscopy (SEM). Magnetic properties were confirmed using Atomic Force Microscopy/Magnetic Force Microscopy (AFM/MFM) and magnetization measurements. The resulting products are negatively charged, rounded in shape and exhibit the superparamagnetic properties what implies their potential applications in engineering and biomedicine areas

Cell surface clustering of Cadherin adhesion complex induced by antibody coated beads

Institute of Scientific and Technical Information of China (English)

无

2000-01-01

Cadherin receptors mediate cell-cell adhesion, signal transduction and assembly of cytoskeletons. How a single transmembrane molecule Cadherin can be involved in multiple functions through modulating its binding activities with many membrane adhesion molecules and cytoskeletal components is an unanswered question which can be elucidated by clues from bead experiments. Human lung cells expressing N-Cadherin were examined. After co-incubation with anti-N-Cadherin monoclonal antibody coated beads, cell surface clustering of N-Cadherin was induced. Immunofluorescent detection demonstrated that in addition to Cadherin, β-Catenin, α-Catenin, α-Actinin and Actin fluorescence also aggregated respectively at the membrane site of bead attachment. Myosin heavy chain (MHC), another major component of Actin cytoskeleton, did not aggregate at the membrane site of bead attachment. Adhesion unrelated protein Con A and polylysine conjugated beads did not induce the clustering of adhesion molecules. It is indicated that the Cadherin/Catenins/α-Actinin/Actin complex is formed at Cadherin mediated cell adherens junction; occupancy and cell surface clustering of Cadherin is crucial for the formation of Cadherin adhesion protein complexes.

Polyaniline coated magnetic carboxymethylcellulose beads for selective removal of uranium ions from aqueous solution

International Nuclear Information System (INIS)

Yakup Arica, M.; Gulay Bayramoglu; Gazi University, Ankara

2016-01-01

Magnetic carboxymethylcellulose (CMC) beads were prepared via ionic cross-linking. Then CMC beads were coated with polyaniline (PANI). The results of FTIR, SEM, DTA and N 2 adsorption-desorption demonstrated that aniline was successfully grafted onto the surface of mCMC beads. The potential use of the prepared adsorbent was used for removal of U(VI) ions. The determined maximum adsorption capacities of the mCMC and mCMC-g-PANI beads for U(VI) ions were 129.4 and 386.5 mg/g dry weight at pH 4.5, respectively. The thermodynamic parameters (ΔH°, ΔS° and ΔG°) were evaluated. The adsorbent systems were regenerated using 0.1 M HCl. (author)

Manipulation of Superparamagnetic Beads on Patterned Exchange-Bias Layer Systems for Biosensing Applications

Directory of Open Access Journals (Sweden)

Arno Ehresmann

2015-11-01

Full Text Available A technology platform based on a remotely controlled and stepwise transport of an array arrangement of superparamagnetic beads (SPB for efficient molecular uptake, delivery and accumulation in the context of highly specific and sensitive analyte molecule detection for the application in lab-on-a-chip devices is presented. The near-surface transport of SPBs is realized via the dynamic transformation of the SPBs’ magnetic potential energy landscape above a magnetically stripe patterned Exchange-Bias (EB thin film layer systems due to the application of sub-mT external magnetic field pulses. In this concept, the SPB velocity is dramatically influenced by the magnitude and gradient of the magnetic field landscape (MFL above the magnetically stripe patterned EB substrate, the SPB to substrate distance, the magnetic properties of both the SPBs and the EB layer system, respectively, as well as by the properties of the external magnetic field pulses and the surrounding fluid. The focus of this review is laid on the specific MFL design in EB layer systems via light-ion bombardment induced magnetic patterning (IBMP. A numerical approach is introduced for the theoretical description of the MFL in comparison to experimental characterization via scanning Hall probe microscopy. The SPB transport mechanism will be outlined in terms of the dynamic interplay between the EB substrate’s MFL and the pulse scheme of the external magnetic field.

Manipulation of Superparamagnetic Beads on Patterned Exchange-Bias Layer Systems for Biosensing Applications.

Science.gov (United States)

Ehresmann, Arno; Koch, Iris; Holzinger, Dennis

2015-11-13

A technology platform based on a remotely controlled and stepwise transport of an array arrangement of superparamagnetic beads (SPB) for efficient molecular uptake, delivery and accumulation in the context of highly specific and sensitive analyte molecule detection for the application in lab-on-a-chip devices is presented. The near-surface transport of SPBs is realized via the dynamic transformation of the SPBs' magnetic potential energy landscape above a magnetically stripe patterned Exchange-Bias (EB) thin film layer systems due to the application of sub-mT external magnetic field pulses. In this concept, the SPB velocity is dramatically influenced by the magnitude and gradient of the magnetic field landscape (MFL) above the magnetically stripe patterned EB substrate, the SPB to substrate distance, the magnetic properties of both the SPBs and the EB layer system, respectively, as well as by the properties of the external magnetic field pulses and the surrounding fluid. The focus of this review is laid on the specific MFL design in EB layer systems via light-ion bombardment induced magnetic patterning (IBMP). A numerical approach is introduced for the theoretical description of the MFL in comparison to experimental characterization via scanning Hall probe microscopy. The SPB transport mechanism will be outlined in terms of the dynamic interplay between the EB substrate's MFL and the pulse scheme of the external magnetic field.

On-Chip Manipulation of Protein-Coated Magnetic Beads via Domain-Wall Conduits

DEFF Research Database (Denmark)

Donolato, Marco; Vavassori, Paolo; Gobbi, Marco

2010-01-01

Geometrically constrained magnetic domain walls (DWs) in magnetic nanowires can be manipulated at the nanometer scale. The inhomogeneous magnetic stray field generated by a DW can capture a magnetic nanoparticle in solution. On-chip nanomanipulation of individual magnetic beads coated with proteins...

Comparison of non-magnetic and magnetic beads in bead-based assays

NARCIS (Netherlands)

Hansenová Maňásková, S.; van Belkum, A.; Endtz, H.P.; Bikker, F.J.; Veerman, E.C.I.; van Wamel, W.J.B.

2016-01-01

Multiplex bead-based flow cytometry is an attractive way for simultaneous, rapid and cost-effective analysis of multiple analytes in a single sample. Previously, we developed various bead-based assays using non-magnetic beads coated with Staphylococcus aureus and Streptococcus pneumoniae antigens

In vitro evaluation of chitosan coated- and uncoated-calcium alginate beads containing methyl salicylate-lactose physical mixture.

Science.gov (United States)

Tamilvanan, S; Karmegam, S

2012-01-01

Methyl salicylate-lactose physical mixture (1:1 and 1:1.5 ratios) was incorporated into calcium alginate beads by a coacervation method involving an ionotropic gelation/polyelectrolyte complexation approach. This study aims to determine the influence of chitosan coating over the beads on drug entrapment efficiency (DEE) and release characteristics in artificial saliva compared to that of the uncoated beads. Changes in formulation parameters (gelation time, concentrations of Ca(2+) and alginate) resulted in decrease in DEE of chitosan-uncoated beads (p methyl salicylate-lactose physical mixture.

Flow-orthogonal bead oscillation in a microfluidic chip with a magnetic anisotropic flux-guide array

DEFF Research Database (Denmark)

Van Pelt, Stijn; Derks, Roy; Matteucci, Marco

2011-01-01

A new concept for the manipulation of superparamagnetic beads inside a microfluidic chip is presented in this paper. The concept allows for bead actuation orthogonal to the flow direction inside a microchannel. Basic manipulation functionalities were studied by means of finite element simulations...

On-chip measurement of the Brownian relaxation frequency of magnetic beads using magnetic tunneling junctions

DEFF Research Database (Denmark)

Donolato, M.; Sogne, E.; Dalslet, Bjarke Thomas

2011-01-01

We demonstrate the detection of the Brownian relaxation frequency of 250 nm diameter magnetic beads using a lab-on-chip platform based on current lines for exciting the beads with alternating magnetic fields and highly sensitive magnetic tunnel junction (MTJ) sensors with a superparamagnetic free...

Decoupling Internalization, Acidification and Phagosmal-Endosomal/Iysosomal Phagocytosis of Internalin A coated Beads in epithelial cells

Energy Technology Data Exchange (ETDEWEB)

Blanchette, C D; Woo, Y; Thomas, C; Shen, N; Sulchek, T A; Hiddessen, A L

2008-12-22

Phagocytosis has been extensively examined in 'professional' phagocytic cells using pH sensitive dyes. However, in many of the previous studies, a separation between the end of internalization, beginning of acidification and completion of phagosomal-endosomal/lysosomal fusion was not clearly established, and in several cases, it was treated as a one-step process. In addition, very little work has been done to systematically examine phagosomal maturation in 'non-professional' phagocytic cells, such as epithelial cells. Therefore, in this study, we developed a simple and novel method to decouple and accurately measure particle internalization, phagosomal acidification and phagosomal-endosomal/lysosomal fusion in Madin-Darby Canine Kidney (MDCK) and Caco-2 epithelial cells. Our method was developed using a pathogen mimetic system consisting of polystyrene beads coated with Internalin A (InlA), a membrane surface protein from Listeria monocytogenes known to trigger receptor-mediated internalization. We achieved independent measurements of the rates of internalization, phagosomal acidification and phagosomal-endosomal/lysosomal fusion in epithelial cells by combining the InlA-coated beads (InlA-beads) with antibody quenching, pH sensitive dyes and endosomal/lysosomal dyes, as follows: the rate of InlA bead internalization was measured via antibody quenching of a pH independent dye (Alexa488) conjugated to InlA-beads, the rate at which phagosomes containing internalized InlA beads became acidified was measured using a pH dependent dye (FITC) conjugated to the beads and the rate of phagosomal-endosomal/lysosomal fusion was measured using a combination of unlabeled InlA-beads and an endosomal/lysosomal dye. By performing these independent measurements under identical experimental conditions, we were able to decouple the three processes and establish time scales for each. In a separate set of experiments, we also exploited the phagosomal acidification

Comparison between simulation and experimentally observed interactions between two magnetic beads in a fluidic system

Energy Technology Data Exchange (ETDEWEB)

Oduwole, Olayinka, E-mail: olayinka.oduwole@eng.ox.ac.uk; Grob, David Tim, E-mail: tim.grob@eng.ox.ac.uk; Sheard, Steve, E-mail: steve.sheard@eng.ox.ac.uk

2016-06-01

Continuous flow separation of magnetic particles within a microfluidic device could lead to improved performance of magnetic bead-based assays but the undesirable formation of bead clusters reduces its efficiency; this efficiency refers to the ability to separate bound magnetic beads from a mixture of particles. Such agglomerates are formed due to magnetic binding forces while hydrodynamic interactions strongly influence the particles' movement. This paper presents a model for interactions between a pair of equal sized super-paramagnetic beads suspended in water within a uniform magnetic field. To the best of our knowledge, we present for the first time a comparison between simulated trajectories and the beads' movement captured on video; the beads were suspended in a stationary fluid placed within a uniform magnetic field. In conclusion, the model is a good approximation for beads interacting with their nearest neighbours and is able to predict the trajectory pattern of these particles in a magnetic bead-based assay. Predicting the magnetically induced interaction of nearby beads will help in determining the density of beads in an assay and in avoiding agglomeration over a fixed time duration. - Highlights: • We modelled the interactions between a pair of super-paramagnetic beads suspended in water within a uniform magnetic field. • We tracked the movement of the bead pair and captured it on video. • We compared the numerical results with the video data and achieved a good agreement. • We predicted the agglomeration time as a function of the separation distance.

Evaluation of umbilical cord mesenchymal stem cells labeling with superparamagnetic iron oxide nanoparticles coated with dextran and complexed with Poly-L-Lysine

International Nuclear Information System (INIS)

Sibov, Tatiana Tais; Mamani, Javier Bustamante; Pavon, Lorena Favaro; Cardenas, Walter Humberto; Gamarra, Lionel Fernel; Miyaki, Liza Aya Mabuchi; Marti, Luciana Cavalheiro; Sardinha, Luiz Roberto; Oliveira, Daniela Mara de

2012-01-01

Objective: The objective of this study was to evaluate the effect of the labeling of umbilical cord vein derived mesenchymal stem cells with superparamagnetic iron oxide nanoparticles coated with dextran and complexed to a non-viral transfector agent transfector poly-L-lysine. Methods: The labeling of mesenchymal stem cells was performed using the superparamagnetic iron oxide nanoparticles/dextran complexed and not complexed to poly-L-lysine. Superparamagnetic iron oxide nanoparticles/dextran was incubated with poly-L-lysine in an ultrasonic sonicator at 37 deg C for 10 minutes for complex formation superparamagnetic iron oxide nanoparticles/dextran/poly-L-lysine by electrostatic interaction. Then, the mesenchymal stem cells were incubated overnight with the complex superparamagnetic iron oxide nanoparticles/dextran/poly-L-lysine and superparamagnetic iron oxide nanoparticles/dextran. After the incubation period the mesenchymal stem cells were evaluated by internalization of the complex superparamagnetic iron oxide nanoparticles/dextran/polyL-lysine and superparamagnetic iron oxide nanoparticles/dextran by Prussian Blue stain. Cellular viability of labeled mesenchymal stem cells was evaluated by cellular proliferation assay using 5,6-carboxyfluorescein-succinimidyl ester method and apoptosis detection by Annexin V- Propidium Iodide assay. Results: mesenchymal stem cells labeled with superparamagnetic iron oxide nanoparticles/ dextran without poly-L-lysine not internalized efficiently the superparamagnetic iron oxide nanoparticles due to its low presence detected within cells. Mesenchymal stem cells labeled with the complex superparamagnetic iron oxide nanoparticles/dextran/polyL-lysine efficiently internalized the superparamagnetic iron oxide nanoparticles due to greater presence in the cells interior. The viability and apoptosis assays demonstrated that the mesenchymal stem cells labeled and not labeled respectively with the superparamagnetic iron oxide

Quantification of specific bindings of biomolecules by magnetorelaxometry

Directory of Open Access Journals (Sweden)

Steinhoff Uwe

2008-03-01

Full Text Available Abstract The binding reaction of the biomolecules streptavidin and anti-biotin antibody, both labelled by magnetic nanoparticles (MNP, to biotin coated on agarose beads, was quantified by magnetorelaxometry (MRX. Highly sensitive SQUID-based MRX revealed the immobilization of the MNP caused by the biotin-streptavidin coupling. We found that about 85% of streptavidin-functionalised MNP bound specifically to biotin-agarose beads. On the other hand only 20% of antibiotin-antibody functionalised MNP were specifically bound. Variation of the suspension medium revealed in comparison to phosphate buffer with 0.1% bovine serum albumin a slight change of the binding behaviour in human serum, probably due to the presence of functioning (non heated serum proteins. Furthermore, in human serum an additional non-specific binding occurs, being independent from the serum protein functionality. The presented homogeneous bead based assay is applicable in simple, uncoated vials and it enables the assessment of the binding kinetics in a volume without liquid flow. The estimated association rate constant for the MNP-labelled streptavidin is by about two orders of magnitude smaller than the value reported for free streptavidin. This is probably due to the relatively large size of the magnetic markers which reduces the diffusion of streptavidin. Furthermore, long time non-exponential kinetics were observed and interpreted as agglutination of the agarose beads.

Methodology description for detection of cellular uptake of PVA coated superparamagnetic iron oxide nanoparticles (SPION) in synovial cells of sheep

International Nuclear Information System (INIS)

Schoepf, Bernhard; Neuberger, Tobias; Schulze, Katja; Petri, Alke; Chastellain, Matthieu; Hofmann, Margarete; Hofmann, Heinrich; Rechenberg, Brigitte von

2005-01-01

The detection of superparamagnetic iron oxide nanoparticles (SPION) in synoviocytes is reported. Synoviocytes were incubated for 2, 12, 24 and 48 h with 1.5 mg/ml of PVA coated SPION under the influence of magnets (12 h). Particles were well tolerated by the synoviocytes, were easily detected using the Turnbulls and Prussian blue reactions between 12 and 24 h

Methodology description for detection of cellular uptake of PVA coated superparamagnetic iron oxide nanoparticles (SPION) in synovial cells of sheep

Energy Technology Data Exchange (ETDEWEB)

Schoepf, Bernhard [Musculoskeletal Research Unit, Equine Hospital, Vetsuisse Faculty Zurich, University of Zurich, Winterthurerstr. 260, 8057 Zurich (Switzerland); Neuberger, Tobias [Musculoskeletal Research Unit, Equine Hospital, Vetsuisse Faculty Zurich, University of Zurich, Winterthurerstr. 260, 8057 Zurich (Switzerland); Schulze, Katja [Musculoskeletal Research Unit, Equine Hospital, Vetsuisse Faculty Zurich, University of Zurich, Winterthurerstr. 260, 8057 Zurich (Switzerland); Petri, Alke [Powder Technology Laboratory, Institute of Materials, Swiss Federal Institute of Technology Lausanne, EPFL, MX-D Ecublens, 1015 Lausanne (Switzerland); Chastellain, Matthieu [Powder Technology Laboratory, Institute of Materials, Swiss Federal Institute of Technology Lausanne, EPFL, MX-D Ecublens, 1015 Lausanne (Switzerland); Hofmann, Margarete [MatSearch, Ch. Jean Pavillard 14, 1009 Pully (Switzerland); Hofmann, Heinrich [Powder Technology Laboratory, Institute of Materials, Swiss Federal Institute of Technology Lausanne, EPFL, MX-D Ecublens, 1015 Lausanne (Switzerland); Rechenberg, Brigitte von [Musculoskeletal Research Unit, Equine Hospital, Vetsuisse Faculty Zurich, University of Zurich, Winterthurerstr. 260, 8057 Zurich (Switzerland)]. E-mail: bvonrechenberg@vetclinics.unizh.ch

2005-05-15

The detection of superparamagnetic iron oxide nanoparticles (SPION) in synoviocytes is reported. Synoviocytes were incubated for 2, 12, 24 and 48 h with 1.5 mg/ml of PVA coated SPION under the influence of magnets (12 h). Particles were well tolerated by the synoviocytes, were easily detected using the Turnbulls and Prussian blue reactions between 12 and 24 h.

Signal amplification for impedimetric genosensing using gold-streptavidin nanoparticles

Energy Technology Data Exchange (ETDEWEB)

Bonanni, A.; Esplandiu, M.J. [Sensors and Biosensors Group, Department of Chemistry, Universitat Autonoma de Barcelona, Edifici Cn, 08193 Bellaterra, Barcelona (Spain); Valle, M. del [Sensors and Biosensors Group, Department of Chemistry, Universitat Autonoma de Barcelona, Edifici Cn, 08193 Bellaterra, Barcelona (Spain)], E-mail: manel.delvalle@uab.es

2008-04-20

Streptavidin-coated gold nanoparticles (strept-AuNPs) were used in this work to amplify the impedimetric signal generated in a biosensor detecting the DNA hybridization event. Probe oligomer was adsorbed onto a graphite epoxy composite (GEC) electrode surface and the impedance measurement was performed in a solution containing the redox marker ferrocyanide/ferricyanide. The biotinylated complementary oligomer was used as target. The change of interfacial charge transfer resistance (R{sub ct}), experimented by the redox marker, was recorded to confirm the hybrid formation. The addition of strept-AuNPs, binding to the target due to the strong streptavidin-biotin interaction, led to a further increment of R{sub ct} thus obtaining significant signal amplification. Strept-AuNPs on the electrode surface were observed by scanning electron microscopy (SEM) after silver enhancement treatment. A competitive binding assay was also performed using unlabelled DNA target to demonstrate its applicability to real sample analysis.

Magnetic bead manipulation in a sub-microliter fluid volume applicable for biosensing

NARCIS (Netherlands)

Derks, R.J.S.; Wimberger-Friedl, R.; Prins, M.W.J.; Dietzel, A.H.

2007-01-01

Magnetic actuation principles using superparamagnetic beads suspended in a fluid are studied in this paper. An exptl. setup contg. a submicroliter fluid vol. surrounded by four miniaturized electromagnets was designed and fabricated. On the basis of optical velocity measurements, the induced

Breakthrough analysis for water disinfection using silver nanoparticles coated resin beads in fixed-bed column

International Nuclear Information System (INIS)

Mthombeni, Nomcebo H.; Mpenyana-Monyatsi, Lizzy; Onyango, Maurice S.; Momba, Maggie N.B.

2012-01-01

Highlights: ► Performance of silver nanoparticles coated resin in water disinfection is presented. ► Sigmoidal models are used to describe breakthrough curves. ► The performance of the media in water disinfection is affected by process variables. ► Test with environmental water shows the media is effective in water disinfection. - Abstract: This study demonstrates the use of silver nanoparticles coated resin beads in deactivating microbes in drinking water in a column filtration system. The coated resin beads are characterized using X-ray diffraction (XRD), Fourier transform infra-red (FT-IR), scanning electron microscope (SEM), transmission electron microscope (TEM) and energy dispersive spectroscopy (EDS) to confirm the functional groups, morphology and the presence of silver nanoparticles on the surface of the resin. The performance of the coated resin is evaluated as a function of bed mass, initial bacterial concentration and flow rate using Escherichia coli as model microbial contaminant in water. The survival curves of E. coli are expressed as breakthrough curves (BTCs), which are modeled using sigmoidal regression equations to obtain relevant rate parameters. The number of bed volumes processed at breakthrough point and capacity of the bed are used as performance indicators. Results show that performance increases with a decrease in initial bacterial concentration, an increase in flow rate and an increase in bed mass.

Aptamer-Modified Magnetic Beads in Biosensing

Science.gov (United States)

Scheper, Thomas; Walter, Johanna-Gabriela

2018-01-01

Magnetic beads (MBs) are versatile tools for the purification, detection, and quantitative analysis of analytes from complex matrices. The superparamagnetic property of magnetic beads qualifies them for various analytical applications. To provide specificity, MBs can be decorated with ligands like aptamers, antibodies and peptides. In this context, aptamers are emerging as particular promising ligands due to a number of advantages. Most importantly, the chemical synthesis of aptamers enables straightforward and controlled chemical modification with linker molecules and dyes. Moreover, aptamers facilitate novel sensing strategies based on their oligonucleotide nature that cannot be realized with conventional peptide-based ligands. Due to these benefits, the combination of aptamers and MBs was already used in various analytical applications which are summarized in this article. PMID:29601533

An integrated open-cavity system for magnetic bead manipulation.

Science.gov (United States)

Abu-Nimeh, F T; Salem, F M

2013-02-01

Superparamagnetic beads are increasingly used in biomedical assays to manipulate, transport, and maneuver biomaterials. We present a low-cost integrated system designed in bulk CMOS to manipulate and separate biomedical magnetic beads. The system consists of 8 × 8 coil-arrays suitable for single bead manipulation, or collaborative multi-bead manipulation, using pseudo-parallel executions. We demonstrate the flexibility of the design in terms of different coil sizes, DC current levels, and layout techniques. In one array module example, the size of a single coil is 30 μm × 30 μm and the full array occupies an area of 248 μm × 248 μm in 0.5 μm CMOS technology. The programmable DC current source supports 8 discrete levels up to 1.5 mA. The total power consumption of the entire module is 9 mW when running at full power.

Radioimmunotargeting with modified streptavidin-biotin. Final report

International Nuclear Information System (INIS)

Cantor, C.R.

1998-01-01

For the past three years, the author has designed and produced several useful streptavidin variants, characterized their properties both in vitro and in vivo, and developed methods and strategies needed for characterization of each molecule and the entire radioimmunotargeting approaches using these molecules. In molecular design of these streptavidin mutants, an enhanced molecular modeling method, including empirical free energy calculations was used which is of great assistance in designing streptavidin mutants effectively and intelligently. When the original proposal was submitted in 1992, the reviewers were rather skeptical about the proposal to design successfully a various streptavidin mutants, particularly a dimeric streptavidin and multi-flavor streptavidins. These were clearly very challenging tasks, but now the author has the molecules in hand. This demonstrates the power of the molecular modeling method he is using and suggests the feasibility of further enhancements of these and other streptavidin variants by the molecular modeling capability. The author has also developed signal amplification methods which could introduce large amounts of radioisotopes specifically at target tumor sites in the radioimmunotargeting approaches

Diffraction-based BioCD biosensor for point-of-care diagnostics

Science.gov (United States)

Choi, H.; Chang, C.; Savran, C.; Nolte, D.

2018-02-01

The BioCD platform technology uses spinning-disk interferometry to detect molecular binding to target molecular probes in biological samples. Interferometric configurations have included differential phase contrast and in-line quadrature detection. For the detection of extremely low analyte concentrations, nano- or microparticles can enhance the signal through background-free diffraction detection. Diffraction signal measurements on BioCD biosensors are achieved by forming gratings on a disc surface. The grating pattern was printed with biotinylated bovine serum albumin (BSA) and streptavidin coated beads were deployed. The diameter of the beads was 1 micron and strong protein bonding occurs between BSA and streptavidin-coated beads at the printed location. The wavelength for the protein binding detection was 635 nm. The periodic pattern on the disc amplified scattered light into the first-order diffraction position. The diffracted signal contains Mie scattering and a randomly-distributed-bead noise contributions. Variation of the grating pattern periodicity modulates the diffraction efficiency. To test multiple spatial frequencies within a single scan, we designed a fan-shaped grating to perform frequency filter multiplexing on a diffraction-based BioCD.

Structure-Guided Design of an Engineered Streptavidin with Reusability to Purify Streptavidin-Binding Peptide Tagged Proteins or Biotinylated Proteins

OpenAIRE

Wu, Sau-Ching; Wong, Sui-Lam

2013-01-01

Development of a high-affinity streptavidin-binding peptide (SBP) tag allows the tagged recombinant proteins to be affinity purified using the streptavidin matrix without the need of biotinylation. The major limitation of this powerful technology is the requirement to use biotin to elute the SBP-tagged proteins from the streptavidin matrix. Tight biotin binding by streptavidin essentially allows the matrix to be used only once. To address this problem, differences in interactions of biotin an...

Superparamagnetic microbead transport induced by a magnetic field on large-area magnetic antidot arrays

Science.gov (United States)

Ouk, Minae; Beach, Geoffrey S. D.

2017-12-01

A method is presented for directed transport of superparamagnetic microbeads (SPBs) on magnetic antidot patterned substrates by applying a rotating elliptical magnetic field. We find a critical frequency for transport, beyond which the bead dynamics transitions from stepwise locomotion to local oscillation. We also find that the out-of-plane (HOOP) and in-plane (HIP) field magnitudes play crucial roles in triggering bead motion. Namely, we find threshold values in HOOP and HIP that depend on bead size, which can be used to independently and remotely address specific bead populations in a multi-bead mixture. These behaviors are explained in terms of the dynamic potential energy lansdscapes computed from micromagnetic simulations of the substrate magnetization configuration. Finally, we show that large-area magnetic patterns suitable for particle transport and sorting can be fabricated through a self-assembly lithography technique, which provides a simple, cost-effective means to integrate magnetic actuation into microfluidic systems.

Streptavidin-coated gold nanoparticles: critical role of oligonucleotides on stability and fractal aggregation

Directory of Open Access Journals (Sweden)

Roberta D'Agata

2017-01-01

Full Text Available Gold nanoparticles (AuNPs exhibit unique properties that can be modulated through a tailored surface functionalization, enabling their targeted use in biochemical sensing and medical diagnostics. In particular, streptavidin-modified AuNPs are increasingly used for biosensing purposes. We report here a study of AuNPs surface-functionalized with streptavidin-biotinylated oligonucleotide, focussing on the role played by the oligonucleotide probes in the stabilization/destabilization of the functionalized nanoparticle dispersion. The behaviour of the modified AuNP dispersion as a consequence of the competitive displacement of the biotinylated oligonucleotide has been investigated and the critical role of displaced oligonucletides in triggering the quasi one-dimensional aggregation of nanoparticles is demonstrated for the first time. The thorough understanding of the fundamental properties of bioconjugated AuNPs is of great importance for the design of highly sensitive and reliable functionalized AuNP-based assays.

Flow Cytometry-Based Bead-Binding Assay for Measuring Receptor Ligand Specificity

NARCIS (Netherlands)

Sprokholt, Joris K.; Hertoghs, Nina; Geijtenbeek, Teunis B. H.

2016-01-01

In this chapter we describe a fluorescent bead-binding assay, which is an efficient and feasible method to measure interaction between ligands and receptors on cells. In principle, any ligand can be coated on fluorescent beads either directly or via antibodies. Binding between ligand-coated beads

Mechanism of Dimercaptosuccinic Acid Coated Superparamagnetic Iron Oxide Nanoparticles with Human Serum Albumin.

Science.gov (United States)

Zhao, Lining; Song, Wei; Wang, Jing; Yan, Yunxing; Chen, Jiangwei; Liu, Rutao

2015-12-01

To research the mechanism of dimercaptosuccinic acid coated-superparamagnetic iron oxide nanoparticles (SPION) with human serum albumin (HSA), the methods of spectroscopy, molecular modeling calculation, and calorimetry were used in this paper. The inner filter effect of the fluorescence intensity was corrected to obtain the accurate results. Ultraviolet-visible absorption and circular dichroism spectra reflect that SPION changed the secondary structure with a loss of α-helix and loosened the protein skeleton of HSA; the activity of the protein was also affected by the increasing exposure of SPION. Fluorescence lifetime measurement indicates that the quenching mechanism type of this system was static quenching. The isothermal titration calorimetry measurement and molecular docking calculations prove that the predominant force of this system was the combination of Van der Waals' force and hydrogen bonds. © 2015 Wiley Periodicals, Inc.

One-step isolation of plasma membrane proteins using magnetic beads with immobilized concanavalin A

DEFF Research Database (Denmark)

Lee, Yu-Chen; Block, Gregory; Chen, Huiwen

2008-01-01

We have developed a simple method for isolating and purifying plasma membrane proteins from various cell types. This one-step affinity-chromatography method uses the property of the lectin concanavalin A (ConA) and the technique of magnetic bead separation to obtain highly purified plasma membrane...... proteins from crude membrane preparations or cell lines. ConA is immobilized onto magnetic beads by binding biotinylated ConA to streptavidin magnetic beads. When these ConA magnetic beads were used to enrich plasma membranes from a crude membrane preparation, this procedure resulted in 3.7-fold enrichment...... of plasma membrane marker 5'-nucleotidase activity with 70% recovery of the activity in the crude membrane fraction of rat liver. In agreement with the results of 5'-nucleotidase activity, immunoblotting with antibodies specific for a rat liver plasma membrane protein, CEACAM1, indicated that CEACAM1...

Improved Peak Capacity for Capillary Electrophoretic Separations of Enzyme Inhibitors with Activity-Based Detection Using Magnetic Bead Microreactors

Science.gov (United States)

Yan, Xiaoyan; Gilman, S. Douglass

2010-01-01

A technique for separating and detecting enzyme inhibitors was developed using capillary electrophoresis with an enzyme microreactor. The on-column enzyme microreactor was constructed using NdFeB magnet(s) to immobilize alkaline phosphatase-coated superparamagnetic beads (2.8 μm diameter) inside a capillary before the detection window. Enzyme inhibition assays were performed by injecting a plug of inhibitor into a capillary filled with the substrate, AttoPhos. Product generated in the enzyme microreactor was detected by laser-induced fluorescence. Inhibitor zones electrophoresed through the capillary, passed through the enzyme microreactor, and were observed as negative peaks due to decreased product formation. The goal of this study was to improve peak capacities for inhibitor separations relative to previous work, which combined continuous engagement electrophoretically mediated microanalysis (EMMA) and transient engagement EMMA to study enzyme inhibition. The effects of electric field strength, bead injection time and inhibitor concentrations on peak capacity and peak width were investigated. Peak capacities were increased to ≥20 under optimal conditions of electric field strength and bead injection time for inhibition assays with arsenate and theophylline. Five reversible inhibitors of alkaline phosphatase (theophylline, vanadate, arsenate, L-tryptophan and tungstate) were separated and detected to demonstrate the ability of this technique to analyze complex inhibitor mixtures. PMID:20024913

Enzyme-linked electrochemical DNA ligation assay using magnetic beads.

Science.gov (United States)

Stejskalová, Eva; Horáková, Petra; Vacek, Jan; Bowater, Richard P; Fojta, Miroslav

2014-07-01

DNA ligases are essential enzymes in all cells and have been proposed as targets for novel antibiotics. Efficient DNA ligase activity assays are thus required for applications in biomedical research. Here we present an enzyme-linked electrochemical assay based on two terminally tagged probes forming a nicked junction upon hybridization with a template DNA. Nicked DNA bearing a 5' biotin tag is immobilized on the surface of streptavidin-coated magnetic beads, and ligated product is detected via a 3' digoxigenin tag recognized by monoclonal antibody-alkaline phosphatase conjugate. Enzymatic conversion of napht-1-yl phosphate to napht-1-ol enables sensitive detection of the voltammetric signal on a pyrolytic graphite electrode. The technique was tested under optimal conditions and various situations limiting or precluding the ligation reaction (such as DNA substrates lacking 5'-phosphate or containing a base mismatch at the nick junction, or application of incompatible cofactor), and utilized for the analysis of the nick-joining activity of a range of recombinant Escherichia coli DNA ligase constructs. The novel technique provides a fast, versatile, specific, and sensitive electrochemical assay of DNA ligase activity.

The Drug Release Profile from Calcium-induced Alginate Gel Beads Coated with an Alginate Hydrolysate

Directory of Open Access Journals (Sweden)

Susumu Kawashima

2007-11-01

Full Text Available Calcium-induced alginate gel bead (Alg-Ca coated with an alginate hydrolysate(Alg, e.g. the guluronic acid block (GB was prepared and the model drug, hydrocortisonerelease profiles were investigated under simulated gastrointestinal conditions. Theirmolecular weights were one sixth or one tenth that of Alg and the diffraction patterns of thehydrolysates resembled that of Alg. The drug release rate from Alg-Ca coated with GBapparently lowered than that of Alg-Ca (coating-free in the gastric juice (pH1.2. And thecoating did not resist the disintegration of Alg-Ca in the intestinal juice (pH 6.8 and thegel erosion accelerated the drug release. On the other hand, for the coated Alg-Cacontaining chitosan, the drug release showed zero-order kinetics without rapid erosion ofAlg-Ca. The drug release rate from Alg-Ca was able to be controlled by the coating andmodifying the composition of the gel matrix.

A multi-purpose ultrasonic streaming mixer for integrated magnetic bead ELISAs

International Nuclear Information System (INIS)

Brandhoff, Lukas; Lang, Walter; Vellekoop, Michael J; Zirath, Helene; Peham, Johannes; Wiesinger-Mayr, Herbert; Salas, Mariugenia; Haller, Anna; Spittler, Andreas; Schnetz, Guntram

2015-01-01

We present an ultrasonic streaming mixer for disposable and on-chip magnetic bead ELISAs. The ultrasonic transducer is placed at system-level to keep cost per chip as low as possible, and is coupled to the chip by means of a solid ultrasonic horn. The system provides mixing of liquids, as well as dispersion of the superparamagnetic beads in the ELISA. Additionally it can be used clean the chamber surface from nonspecifically bound proteins during the washing steps in the ELISA protocol. Using our system the time for the ELISA protocol has been greatly reduced down to 30 min. (paper)

Multifunctional PEG-carboxylate copolymer coated superparamagnetic iron oxide nanoparticles for biomedical application

Science.gov (United States)

Illés, Erzsébet; Szekeres, Márta; Tóth, Ildikó Y.; Szabó, Ákos; Iván, Béla; Turcu, Rodica; Vékás, Ladislau; Zupkó, István; Jaics, György; Tombácz, Etelka

2018-04-01

Biocompatible magnetite nanoparticles (MNPs) were prepared by post-coating the magnetic nanocores with a synthetic polymer designed specifically to shield the particles from non-specific interaction with cells. Poly(ethylene glycol) methyl ether methacrylate (PEGMA) macromonomers and acrylic acid (AA) small molecular monomers were chemically coupled by quasi-living atom transfer radical polymerization (ATRP) to a comb-like copolymer, P(PEGMA-co-AA) designated here as P(PEGMA-AA). The polymer contains pendant carboxylate moieties near the backbone and PEG side chains. It is able to bind spontaneously to MNPs; stabilize the particles electrostatically via the carboxylate moieties and sterically via the PEG moieties; provide high protein repellency via the structured PEG layer; and anchor bioactive proteins via peptide bond formation with the free carboxylate groups. The presence of the P(PEGMA-AA) coating was verified in XPS experiments. The electrosteric (i.e., combined electrostatic and steric) stabilization is efficient down to pH 4 (at 10 mM ionic strength). Static magnetization and AC susceptibility measurements showed that the P(PEGMA-AA)@MNPs are superparamagnetic with a saturation magnetization value of 55 emu/g and that both single core nanoparticles and multicore structures are present in the samples. The multicore components make our product well suited for magnetic hyperthermia applications (SAR values up to 17.44 W/g). In vitro biocompatibility, cell internalization, and magnetic hyperthermia studies demonstrate the excellent theranostic potential of our product.

Expression of streptavidin gene in bacteria and plants

International Nuclear Information System (INIS)

Guan, Xueni; Wurtele, E.S.; Nikolau, B.J.

1990-01-01

Six biotin-containing proteins are present in plants, representing at least four different biotin enzymes. The physiological function of these biotin enzymes is not understood. Streptavidin, a protein from Streptomyces avidinii, binds tightly and specifically to biotin causing inactivation of biotin enzymes. One approach to elucidating the physiological function of biotin enzymes in plant metabolism is to create transgenic plants expressing the streptavidin gene. A plasmid containing a fused streptavidin-beta-galactosidase gene has been expressed in E. coli. We also have constructed various fusion genes that include an altered CaMV 35S promoter, signal peptides to target the streptavidin protein to specific organelles, and the streptavidin coding gene. We are examining the expression of these genes in cells of carrot

Optimal size for heating efficiency of superparamagnetic dextran-coated magnetite nanoparticles for application in magnetic fluid hyperthermia

Science.gov (United States)

Shaterabadi, Zhila; Nabiyouni, Gholamreza; Soleymani, Meysam

2018-06-01

Dextran-coated magnetite (Fe3O4) nanoparticles with average particle sizes of 4 and 19 nm were synthesized through in situ and semi-two-step co-precipitation methods, respectively. The experimental results confirm the formation of pure phase of magnetite as well as the presence of dextran layer on the surface of modified magnetite nanoparticles. The results also reveal that both samples have the superparamagnetic behavior. Furthermore, calorimetric measurements show that the dextran-coated Fe3O4 nanoparticles with an average size of 4 nm cannot produce any appreciable heat under a biologically safe alternating magnetic field used in hyperthermia therapy; whereas, the larger ones (average size of 19 nm) are able to increase the temperature of their surrounding medium up to above therapeutic range. In addition, measured specific absorption rate (SAR) values confirm that magnetite nanoparticles with an average size of 19 nm are very excellent candidates for application in magnetic hyperthermia therapy.

Plasma membrane isolation using immobilized concanavalin A magnetic beads.

Science.gov (United States)

Lee, Yu-Chen; Srajer Gajdosik, Martina; Josic, Djuro; Lin, Sue-Hwa

2012-01-01

Isolation of highly purified plasma membranes is the key step in constructing the plasma membrane proteome. Traditional plasma membrane isolation method takes advantage of the differential density of organelles. While differential centrifugation methods are sufficient to enrich for plasma membranes, the procedure is lengthy and results in low recovery of the membrane fraction. Importantly, there is significant contamination of the plasma membranes with other organelles. The traditional agarose affinity matrix is suitable for isolating proteins but has limitation in separating organelles due to the density of agarose. Immobilization of affinity ligands to magnetic beads allows separation of affinity matrix from organelles through magnets and could be developed for the isolation of organelles. We have developed a simple method for isolating plasma membranes using lectin concanavalin A (ConA) magnetic beads. ConA is immobilized onto magnetic beads by binding biotinylated ConA to streptavidin magnetic beads. The ConA magnetic beads are used to bind glycosylated proteins present in the membranes. The bound membranes are solubilized from the magnetic beads with a detergent containing the competing sugar alpha methyl mannoside. In this study, we describe the procedure of isolating rat liver plasma membranes using sucrose density gradient centrifugation as described by Neville. We then further purify the membrane fraction by using ConA magnetic beads. After this purification step, main liver plasma membrane proteins, especially the highly glycosylated ones and proteins containing transmembrane domains could be identified by LC-ESI-MS/MS. While not described here, the magnetic bead method can also be used to isolate plasma membranes from cell lysates. This membrane purification method should expedite the cataloging of plasma membrane proteome.

SEPARATION OF CELL POPULATIONS BY SUPER-PARAMAGNETIC PARTICLES WITH CONTROLLED SURFACE FUNCTIONALITY

Directory of Open Access Journals (Sweden)

Lootsik M. D.

2014-02-01

Full Text Available The recognition and isolation of specific mammalian cells by the biocompatible polymer coated super-paramagnetic particles with determined surface functionality were studied. The method of synthesis of nanoscaled particles on a core of iron III oxide (Fe2O3, magemit coated with a polymer shell containing reactive oligoperoxide groups for attachment of ligands is described. By using the developed superparamagnetic particles functionalized with peanut agglutinin (PNA we have separated the sub-populations of PNA+ and PNA– cells from ascites of murine Nemeth-Kellner lymphoma. In another type of experiment, the particles were opsonized with proteins of the fetal calf serum that improved biocompatibility of the particles and their ingestion by cultivated murine macrophages J774.2. Macrophages loaded with the particles were effeciently separated from the particles free cells by using the magnet. Thus, the developed surface functionalized superparamagnetic particles showed to be a versatile tool for cell separation independent on the mode of particles’ binding with cell surface or their engulfment by the targeted cells.

Structure-guided design of an engineered streptavidin with reusability to purify streptavidin-binding peptide tagged proteins or biotinylated proteins.

Directory of Open Access Journals (Sweden)

Sau-Ching Wu

Full Text Available Development of a high-affinity streptavidin-binding peptide (SBP tag allows the tagged recombinant proteins to be affinity purified using the streptavidin matrix without the need of biotinylation. The major limitation of this powerful technology is the requirement to use biotin to elute the SBP-tagged proteins from the streptavidin matrix. Tight biotin binding by streptavidin essentially allows the matrix to be used only once. To address this problem, differences in interactions of biotin and SBP with streptavidin were explored. Loop3-4 which serves as a mobile lid for the biotin binding pocket in streptavidin is in the closed state with biotin binding. In contrast, this loop is in the open state with SBP binding. Replacement of glycine-48 with a bulkier residue (threonine in this loop selectively reduces the biotin binding affinity (Kd from 4 × 10(-14 M to 4.45 × 10(-10 M without affecting the SBP binding affinity. Introduction of a second mutation (S27A to the first mutein (G48T results in the development of a novel engineered streptavidin SAVSBPM18 which could be recombinantly produced in the functional form from Bacillus subtilis via secretion. To form an intact binding pocket for tight binding of SBP, two diagonally oriented subunits in a tetrameric streptavidin are required. It is vital for SAVSBPM18 to be stably in the tetrameric state in solution. This was confirmed using an HPLC/Laser light scattering system. SAVSBPM18 retains high binding affinity to SBP but has reversible biotin binding capability. The SAVSBPM18 matrix can be applied to affinity purify SBP-tagged proteins or biotinylated molecules to homogeneity with high recovery in a reusable manner. A mild washing step is sufficient to regenerate the matrix which can be reused for multiple rounds. Other applications including development of automated protein purification systems, lab-on-a-chip micro-devices, reusable biosensors, bioreactors and microarrays, and strippable

Structure-guided design of an engineered streptavidin with reusability to purify streptavidin-binding peptide tagged proteins or biotinylated proteins.

Science.gov (United States)

Wu, Sau-Ching; Wong, Sui-Lam

2013-01-01

Development of a high-affinity streptavidin-binding peptide (SBP) tag allows the tagged recombinant proteins to be affinity purified using the streptavidin matrix without the need of biotinylation. The major limitation of this powerful technology is the requirement to use biotin to elute the SBP-tagged proteins from the streptavidin matrix. Tight biotin binding by streptavidin essentially allows the matrix to be used only once. To address this problem, differences in interactions of biotin and SBP with streptavidin were explored. Loop3-4 which serves as a mobile lid for the biotin binding pocket in streptavidin is in the closed state with biotin binding. In contrast, this loop is in the open state with SBP binding. Replacement of glycine-48 with a bulkier residue (threonine) in this loop selectively reduces the biotin binding affinity (Kd) from 4 × 10(-14) M to 4.45 × 10(-10) M without affecting the SBP binding affinity. Introduction of a second mutation (S27A) to the first mutein (G48T) results in the development of a novel engineered streptavidin SAVSBPM18 which could be recombinantly produced in the functional form from Bacillus subtilis via secretion. To form an intact binding pocket for tight binding of SBP, two diagonally oriented subunits in a tetrameric streptavidin are required. It is vital for SAVSBPM18 to be stably in the tetrameric state in solution. This was confirmed using an HPLC/Laser light scattering system. SAVSBPM18 retains high binding affinity to SBP but has reversible biotin binding capability. The SAVSBPM18 matrix can be applied to affinity purify SBP-tagged proteins or biotinylated molecules to homogeneity with high recovery in a reusable manner. A mild washing step is sufficient to regenerate the matrix which can be reused for multiple rounds. Other applications including development of automated protein purification systems, lab-on-a-chip micro-devices, reusable biosensors, bioreactors and microarrays, and strippable detection agents for

Preparation of ionic-crosslinked chitosan-based gel beads and effect of reaction conditions on drug release behaviors.

Science.gov (United States)

Chen, Shilan; Liu, Mingzhu; Jin, Shuping; Wang, Bin

2008-02-12

Drug-loaded chitosan (CS) beads were prepared under simple and mild condition using trisodium citrate as ionic crosslinker. The beads were further coated with poly(methacrylic acid) (PMAA) by dipping the beads in PMAA aqueous solution. The surface and cross-section morphology of these beads were observed by scanning electron microscopy and the observation showed that the coating beads had core-shell structure. In vitro release of model drug from these beads obtained under different reaction conditions was investigated in buffer medium (pH 1.8). The results showed that the rapid drug release was restrained by PMAA coating and the optimum conditions for preparing CS-based drug-loaded beads were decided through the effect of reaction conditions on the drug release behaviors. In addition, the drug release mechanism of CS-based drug-loaded beads was analyzed by Peppa's potential equation. According to this study, the ionic-crosslinked CS beads coated by PMAA could serve as suitable candidate for drug site-specific carrier in stomach.

Near-infrared-responsive, superparamagnetic Au@Co nanochains

Directory of Open Access Journals (Sweden)

Varadee Vittur

2017-08-01

Full Text Available This manuscript describes a new type of nanomaterial, namely superparamagnetic Au@Co nanochains with optical extinctions in the near infrared (NIR. The Au@Co nanochains were synthesized via a one-pot galvanic replacement route involving a redox-transmetalation process in aqueous medium, where Au salt was reduced to form Au shells on Co seed templates, affording hollow Au@Co nanochains. The Au shells serve not only as a protective coating for the Co nanochain cores, but also to give rise to the optical properties of these unique nanostructures. Importantly, these bifunctional, magneto-optical Au@Co nanochains combine the advantages of nanophotonics (extinction at ca. 900 nm and nanomagnetism (superparamagnetism and provide a potentially useful new nanoarchitecture for biomedical or catalytic applications that can benefit from both activation by light and manipulation using an external magnetic field.

On-chip Brownian relaxation measurements of magnetic nanobeads in the time domain

DEFF Research Database (Denmark)

Østerberg, Frederik Westergaard; Rizzi, Giovanni; Hansen, Mikkel Fougt

2013-01-01

the time and frequency domain methods on Brownian relaxation detection of clustering of streptavidin coated magnetic beads in the presence of different concentrations of biotin-conjugated bovine serum albumin and obtain comparable results. In the time domain, a measurement is carried out in less than 30 s...

Spectroscopic and magnetic studies of highly dispersible superparamagnetic silica coated magnetite nanoparticles

Energy Technology Data Exchange (ETDEWEB)

Tadyszak, Krzysztof [NanoBioMedical Centre, Adam Mickiewicz University, ul. Umultowska 85, 61-614 Poznań (Poland); Institute of Molecular Physics Polish Academy of Sciences, ul. Mariana Smo.luchowskiego 17, 60-179 Poznań (Poland); Kertmen, Ahmet, E-mail: ahmet.kertmen@pg.gda.pl [Department of Pharmaceutical Technology and Biochemistry, Faculty of Chemistry, Gdańsk University of Technology, Narutowicza 11/12, 80-233 Gdańsk (Poland); Coy, Emerson [NanoBioMedical Centre, Adam Mickiewicz University, ul. Umultowska 85, 61-614 Poznań (Poland); Andruszkiewicz, Ryszard; Milewski, Sławomir [Department of Pharmaceutical Technology and Biochemistry, Faculty of Chemistry, Gdańsk University of Technology, Narutowicza 11/12, 80-233 Gdańsk (Poland); Kardava, Irakli; Scheibe, Błażej; Jurga, Stefan [NanoBioMedical Centre, Adam Mickiewicz University, ul. Umultowska 85, 61-614 Poznań (Poland); Chybczyńska, Katarzyna, E-mail: katarzyna.chybczynska@ifmpan.poznan.pl [Institute of Molecular Physics Polish Academy of Sciences, ul. Mariana Smo.luchowskiego 17, 60-179 Poznań (Poland)

2017-07-01

Highlights: • Superparamagnetic core-shell nanoparticles of Fe{sub 2}O{sub 3}@Silica were obtained. • Magnetic response was studied by DC, AC magnetometry and EPR spectroscopy. • Nanoparticles show magnetite structure with a well-defined Verwey transition. • Samples show no inter particle magnetic interactions or agglomeration. - Abstract: Superparamagnetic behavior in aqueously well dispersible magnetite core-shell Fe{sub 3}O{sub 4}@SiO{sub 2} nanoparticles is presented. The magnetic properties of core-shell nanoparticles were measured with use of the DC, AC magnetometry and EPR spectroscopy. Particles where characterized by HR-TEM and Raman spectroscopy, showing a crystalline magnetic core of 11.5 ± 0.12 nm and an amorphous silica shell of 22 ± 1.5 nm in thickness. The DC, AC magnetic measurements confirmed the superparamagnetic nature of nanoparticles, additionally the EPR studies performed at much higher frequency than DC, AC magnetometry (9 GHz) have confirmed the paramagnetic nature of the nanoparticles. Our results show the excellent magnetic behavior of the particles with a clear magnetite structure, which are desirable properties for environmental remediation and biomedical applications.

Evaluation of the Effect of Psyllium on the Viability of Lactobacillus Acidophilus in Alginate-Polyl Lysine Beads.

Science.gov (United States)

Esmaeilzadeh, Jaleh; Nazemiyeh, Hossein; Maghsoodi, Maryam; Lotfipour, Farzaneh

2016-09-01

Purpose: Psylliumseeds are used in traditional herbal medicine to treat various disorders. Moreover, as a soluble fiber, psyllium has potential to stimulate bacterial growth in digestive system. We aimed to substitute alkali-extractable polysaccharides of psyllium for alginate in beads with second coat of poly-l-lysine to coat Lactobacillus acidophilus. Methods: Beads were prepared using extrusion technique. Poly-l-lysine as second coat was incorporated on optimum alginate/psyllium beads using immersion technique. Beads were characterized in terms of size, encapsulation efficiency, integrity and bacterial survival in harsh conditions. Results: Beads with narrow size distribution ranging from 1.85 ± 0.05 to 2.40 ± 0.18 mm with encapsulation efficiency higher than 96% were achieved. Psyllium concentrations in beads did not produce constant trend in bead sizes. Surface topography by SEM showed that substitution of psyllium enhanced integrity of obtained beads. Psyllium successfully protected the bacteria against acidic condition and lyophilization equal to alginate in the beads. Better survivability with beads of alginate/psyllium-poly-l-lysine was achieved with around 2 log rise in bacterial count in acid condition compared to the corresponding single coat beads. Conclusion: Alginate/psyllium (1:2) beads with narrow size distribution and high encapsulation efficiency of the bacteria have been achieved. Presence of psyllium produced a much smoother and integrated surface texture for the beads with sufficient protection of the bacteria against acidic condition as much as alginate. Considering the health benefits of psyllium and its prebiotic activity, psyllium can be beneficially replaced in part for alginate in probiotic coating.

Efficacy of polymer coating of probiotic beads suspended in pressurized and pasteurized longan juices on the exposure to simulated gastrointestinal environment.

Science.gov (United States)

Chaikham, Pittaya; Apichartsrangkoon, Arunee; George, Trevor; Jirarattanarangsri, Wachira

2013-11-01

Alginate-coated Lactobacillus acidophilus LA5 or Lactobacillus casei 01 was recoated with either 0.1-0.5% (w/v) alginate or 0.05-0.15% (w/v) poly-L-lysine (PLL) plus 0.2% (w/v) alginate or 5-15% (w/v) gelatin, after which they were determined for survivability in gastric or bile longan juices. The morphology of encapsulated probiotic cells illustrated that recoated beads with 0.5% alginate showed a more compact surface and a greater protective effect than other recoating materials. The recoated beads with 0.5% alginate and 0.05-0.15% PLL plus 0.2% alginate of both strains showed the highest viability in gastric longan juice. In bile longan juice, only 0.5% alginate showed the best protection for both recoated beads. When considering the storage stability, encapsulated L. acidophilus LA5 exhibited a higher viable count than those of the free cells, whereas L. casei 01 showed equivalent viability of both free and double-coated cells. Based on the impact of pressurization or pasteurization, both processed juices gave rise to equivalent survivability of the probiotic cells during storage.

Chitosan coated alginate-xanthan gum bead enhanced pH and thermotolerance of Lactobacillus plantarum LAB12.

Science.gov (United States)

Fareez, Ismail M; Lim, Siong Meng; Mishra, Rakesh K; Ramasamy, Kalavathy

2015-01-01

The vulnerability of probiotics at low pH and high temperature has limited their optimal use as nutraceuticals. This study addressed these issues by adopting a physicochemical driven approach of incorporating Lactobacillus plantarum LAB12 into chitosan (Ch) coated alginate-xanthan gum (Alg-XG) beads. Characterisation of Alg-XG-Ch, which elicited little effect on bead size and polydispersity, demonstrated good miscibility with improved bead surface smoothness and L. plantarum LAB12 entrapment when compared to Alg, Alg-Ch and Alg-XG. Sequential incubation of Alg-XG-Ch in simulated gastric juice and intestinal fluid yielded high survival rate of L. plantarum LAB12 (95%) at pH 1.8 which in turn facilitated sufficient release of probiotics (>7 log CFU/g) at pH 6.8 in both time- and pH-dependent manner. Whilst minimising viability loss at 75 and 90 °C, Alg-XG-Ch improved storage durability of L. plantarum LAB12 at 4 °C. The present results implied the possible use of L. plantarum LAB12 incorporated in Alg-XG-Ch as new functional food ingredient with health claims. Copyright © 2014 Elsevier B.V. All rights reserved.

Kinase-loaded magnetic beads for sequential in vitro phosphorylation of peptides and proteins.

Science.gov (United States)

Hromadkova, Lenka; Kupcik, Rudolf; Vajrychova, Marie; Prikryl, Petr; Charvatova, Andrea; Jankovicova, Barbora; Ripova, Daniela; Bilkova, Zuzana; Slovakova, Marcela

2018-01-15

Post-translational modifications, including phosphorylation, greatly impact the physiological function of proteins, especially those that are natively unfolded and implicated in many neurodegenerative diseases. However, structural and functional studies of such proteins require fully defined phosphorylation, including those that are not physiological. Thus, the kinases ERK2 and GSK-3β were immobilized to various superparamagnetic beads with carboxylic, aldehyde, Ni 2+ , or Co 3+ functional groups, with a view to efficiently phosphorylate peptides and proteins in vitro. Full phosphorylation of specific synthetic peptides confirmed that beads were successfully loaded with kinases. Remarkably, enzymes covalently immobilized on carboxylated SeraMag beads remained active upon reuse, with residual activity after 10 uses 99.5 ± 0.34% for GSK-3β and 36.2 ± 2.01% for ERK2. The beads were also used to sequentially phosphorylate recombinant tau, which in vivo is a biomarker of Alzheimer's disease. Thus, a system consisting of two fully active kinases immobilized to magnetic beads is demonstrated for the first time. In comparison to soluble enzymes, the beads are easier to handle, reusable, and thus low-cost. Importantly, these beads are also convenient to remove from reactions to minimize contamination of phosphorylated products or to exchange with other kinases.

In vitro transcription of a torsionally constrained template

DEFF Research Database (Denmark)

Bentin, Thomas; Nielsen, Peter E

2002-01-01

of torsionally constrained DNA by free RNAP. We asked whether or not a newly synthesized RNA chain would limit transcription elongation. For this purpose we developed a method to immobilize covalently closed circular DNA to streptavidin-coated beads via a peptide nucleic acid (PNA)-biotin conjugate in principle...

The isolation and characterization of twelve novel microsatellite loci ...

African Journals Online (AJOL)

hp

2013-12-18

Dec 18, 2013 ... MboI adapter2 (5'-GTCAAGAATTCGGTACCGTCGAC-3') were ligated to the digested products using T4 DNA ligase. The ligated product was hybridized with biotin-labeled sequence repeats (SSR) probes. (GT)15, (CT)15, and the hybrid mixture was incubated with magnetic beads coated with streptavidin.

Incorporation and release of drug into/from superparamagnetic iron oxide nanoparticles

Energy Technology Data Exchange (ETDEWEB)

Maver, Uros; Bele, Marjan [National Institute of Chemistry Slovenia, Hajdrihova 19, 1000 Ljubljana (Slovenia); Makovec, Darko; Campelj, Stanislav [Jozef Stefan Institute, Jamova 39, 1000 Ljubljana (Slovenia); Jamnik, Janko [National Institute of Chemistry Slovenia, Hajdrihova 19, 1000 Ljubljana (Slovenia); Gaberscek, Miran [National Institute of Chemistry Slovenia, Hajdrihova 19, 1000 Ljubljana (Slovenia)], E-mail: miran.gaberscek@ki.si

2009-10-15

The aim of this study was to attach a model drug (naproxen) onto superparamagnetic iron oxide nanoparticles (SPION). First, SPION were coated with thin layer of silica that contained micropores. We demonstrated that such surface functionalization could be optimized by the use of citric acid which prevented SPION agglomeration during the procedure. HRTEM investigation showed a uniform 1-2-nm-thick silica coating around SPION. This coating did not affect significantly the magnetic properties of the SPION. Into the coated SPION we successfully incorporated about 30 wt% of naproxen. The latter was readily released after immersion into a testing solution. The composites could be interesting for potential use in diagnostics.

Incorporation and release of drug into/from superparamagnetic iron oxide nanoparticles

International Nuclear Information System (INIS)

Maver, Uros; Bele, Marjan; Makovec, Darko; Campelj, Stanislav; Jamnik, Janko; Gaberscek, Miran

2009-01-01

The aim of this study was to attach a model drug (naproxen) onto superparamagnetic iron oxide nanoparticles (SPION). First, SPION were coated with thin layer of silica that contained micropores. We demonstrated that such surface functionalization could be optimized by the use of citric acid which prevented SPION agglomeration during the procedure. HRTEM investigation showed a uniform 1-2-nm-thick silica coating around SPION. This coating did not affect significantly the magnetic properties of the SPION. Into the coated SPION we successfully incorporated about 30 wt% of naproxen. The latter was readily released after immersion into a testing solution. The composites could be interesting for potential use in diagnostics.

Recent advances in synthesis and surface modification of superparamagnetic iron oxide nanoparticles with silica

Energy Technology Data Exchange (ETDEWEB)

Sodipo, Bashiru Kayode, E-mail: bashirsodipo@gmail.com [School of Physics, Universiti Sains Malaysia, 11800 Pulau Pinang (Malaysia); Nano-Biotechnology Research and Innovation (NanoBRI), Institute for Research in Molecular Medicine (INFORMM), Universiti Sains Malaysia, 11800 Pulau Pinang (Malaysia); Aziz, Azlan Abdul [School of Physics, Universiti Sains Malaysia, 11800 Pulau Pinang (Malaysia); Nano-Biotechnology Research and Innovation (NanoBRI), Institute for Research in Molecular Medicine (INFORMM), Universiti Sains Malaysia, 11800 Pulau Pinang (Malaysia)

2016-10-15

Research on synthesis of superparamagnetic iron oxide nanoparticles (SPION) and its surface modification for biomedical applications is of intense interest. Due to superparamagnetic property of SPION, the nanoparticles have large magnetic susceptibility, single magnetic domain and controllable magnetic behaviour. However, owing to easy agglomeration of SPION, surface modification of the magnetic particles with biocompatible materials such as silica nanoparticle has gained much attention in the last decade. In this review, we present recent advances in synthesis of SPION and various routes of producing silica coated SPION. - Highlights: • We present recent advances in synthesis of SPION and various routes of producing silica coated SPION • The synthetic routes of producing SPION can be classified into three: physical, chemical and biological methods. • The chemical method is the most cited method of producing SPION and it sub-classified into liquid and gas phase. • The techniques of producing silica coated SPION is grouped into seeded and non-seeded methods.

Recent advances in synthesis and surface modification of superparamagnetic iron oxide nanoparticles with silica

International Nuclear Information System (INIS)

Sodipo, Bashiru Kayode; Aziz, Azlan Abdul

2016-01-01

Research on synthesis of superparamagnetic iron oxide nanoparticles (SPION) and its surface modification for biomedical applications is of intense interest. Due to superparamagnetic property of SPION, the nanoparticles have large magnetic susceptibility, single magnetic domain and controllable magnetic behaviour. However, owing to easy agglomeration of SPION, surface modification of the magnetic particles with biocompatible materials such as silica nanoparticle has gained much attention in the last decade. In this review, we present recent advances in synthesis of SPION and various routes of producing silica coated SPION. - Highlights: • We present recent advances in synthesis of SPION and various routes of producing silica coated SPION • The synthetic routes of producing SPION can be classified into three: physical, chemical and biological methods. • The chemical method is the most cited method of producing SPION and it sub-classified into liquid and gas phase. • The techniques of producing silica coated SPION is grouped into seeded and non-seeded methods.

Fluctuations of cytoskeleton-bound microbeads-the effect of bead-receptor binding dynamics

International Nuclear Information System (INIS)

Metzner, C; Raupach, C; Mierke, C T; Fabry, B

2010-01-01

The cytoskeleton (CSK) of living cells is a crosslinked fiber network, subject to ongoing biochemical remodeling processes that can be visualized by tracking the spontaneous motion of CSK-bound microbeads. The bead motion is characterized by anomalous diffusion with a power-law time evolution of the mean square displacement (MSD), and can be described as a stochastic transport process with apparent diffusivity D and power-law exponent β: MSD ∼ D (t/t 0 ) β . Here we studied whether D and β change with the time that has passed after the initial bead-cell contact, and whether they are sensitive to bead coating (fibronectin, integrin antibodies, poly-L-lysine, albumin) and bead size (0.5-4.5 μm). The measurements are interpreted in the framework of a simple model that describes the bead as an overdamped particle coupled to the fluctuating CSK network by an elastic spring. The viscous damping coefficient characterizes the degree of bead internalization into the cell, and the spring constant characterizes the strength of the binding of the bead to the CSK. The model predicts distinctive signatures of the MSD that change with time as the bead couples more tightly to the CSK and becomes internalized. Experimental data show that the transition from the unbound to the tightly bound state occurs in an all-or-nothing manner. The time point of this transition shows considerable variability between individual cells (2-30 min) and depends on the bead size and bead coating. On average, this transition occurs later for smaller beads and beads coated with ligands that trigger the formation of adhesion complexes (fibronectin, integrin antibodies). Once the bead is linked to the CSK, however, the ligand type and bead size have little effect on the MSD. On longer timescales of several hours after bead addition, smaller beads are internalized into the cell more readily, leading to characteristic changes in the MSD that are consistent with increased viscous damping by the

Microchannel conductivity measurements in microchip for on line monitoring of dephosphorylation rates of organic phosphates using paramagnetic-beads linked alkaline phosphatase.

Science.gov (United States)

Kechadi, Mohammed; Sotta, Bruno; Gamby, Jean

2015-01-01

This paper presents the use of polymer coated microelectrodes for the realtime conductivity monitoring in a microchannel photoablated through the polymer without contact. Based on this strategy, a small conductometry sensor has been developed to record in time conductivity variation when an enzymatic reaction occurs through the channel. The rate constant determination, k2, for the dephosphorylation of organic phosphate-alkaline phosphatase-superparamagnetic beads complex using chemically different substrates such as adenosine monoesterphosphate, adenosine diphosphate and adenosine triphosphate was taken as an example to demonstrate selectivity and sensivity of the detection scheme. The k2 value measured for each adenosine phosphate decreases from 39 to 30 s(-1) in proportion with the number (3, 2 and 1) of attached phosphate moiety, thus emphasizing the steric hindrance effect on kinetics. Copyright © 2014 Elsevier B.V. All rights reserved.

Sodium lauryl sulfate impedes drug release from zinc-crosslinked alginate beads: switching from enteric coating release into biphasic profiles.

Science.gov (United States)

Taha, Mutasem O; Nasser, Wissam; Ardakani, Adel; Alkhatib, Hatim S

2008-02-28

The aim of this research is to investigate the effects of sodium lauryl sulfate (SLS) on ionotropically cross-linked alginate beads. Different levels of SLS were mixed with sodium alginate and chlorpheniramine maleate (as loaded model drug). The resulting viscous solutions were dropped onto aqueous solutions of zinc or calcium ions for ionotropic curing. The generated beads were assessed by their drug releasing profiles, infrared and differential scanning colorimetery (DSC) traits. SLS was found to exert profound concentration-dependent impacts on the characteristics of zinc-crosslinked alginate beads such that moderate modifications in the levels of SLS switched drug release from enteric coating-like behavior to a biphasic release modifiable to sustained-release by the addition of minute amounts of xanthan gum. Calcium cross-linking failed to reproduce the same behavior, probably due to the mainly ionic nature of calcium-carboxylate bonds compared to the coordinate character of their zinc-carboxylate counterparts. Apparently, moderate levels of SLS repel water penetration into the beads, and therefore minimize chlorpheniramine release. However, higher SLS levels seem to discourage polymeric cross-linking and therefore allow biphasic drug release.

Development of liposomes entrapped in alginate beads for the treatment of colorectal cancer.

Science.gov (United States)

Bansal, Divya; Gulbake, Arvind; Tiwari, Jyoti; Jain, Sanjay K

2016-01-01

Folic Acid conjugated liposomes encapsulating Oxaliplatin (L-OHP) were entrapped in alginate beads and further coated with Eudragit-S-100 for effective delivery to colon tumors. Liposomes were prepared by cast film method and folic acid was coupled on the surface of liposomes. They were further entrapped in alginate beads which were Eudragit coated for degradation in the colonic region. The prepared beads were characterized for shape and surface morphology, percentage entrapment efficiency and drug release studies. The in vitro drug release was investigated using a USP dissolution paddle type apparatus in different simulated gastrointestinal fluids. In vivo studies of the beads containing free drug, folic acid coupled and uncoupled liposomes bearing L-OHP was administered orally at the dose of 10mg L-OHP/kg body weight to tumor bearing NUDE/SCID mice. γ-Scintigraphic study showed that Eudragit coated alginate beads entered into the colon of Balb/c mice between 4.20 and 4.50h after oral administration. In vivo data showed that folic acid coupled liposomes entrapped in alginate beads delivered 2.82 ± 0.58 and 21.52 ± 2.76 μg L-OHP/g tissues in the colon and tumor after 12h, reflecting its targeting potential to colon and tumor. The results clearly demonstrate that Eudragit coated calcium alginate beads bearing folic acid coupled liposome can be used as a prospective carrier for drug delivery to colon specific tumor. Copyright © 2015 Elsevier B.V. All rights reserved.

Atherosclerotic imaging using 4 types of superparamagnetic iron oxides: New possibilities for mannan-coated particles

Energy Technology Data Exchange (ETDEWEB)

Tsuchiya, Keiko, E-mail: keikot@belle.shiga-medac.jp [Department of Radiology, Shiga University of Medical Science, Setatsukinowa-cho, Otsu, Shiga 520-2192 (Japan); Nitta, Norihisa, E-mail: r34nitta@yahoo.co.jp [Department of Radiology, Shiga University of Medical Science, Setatsukinowa-cho, Otsu, Shiga 520-2192 (Japan); Sonoda, Akinaga, E-mail: akinagasonoda@yahoo.co.jp [Department of Radiology, Shiga University of Medical Science, Setatsukinowa-cho, Otsu, Shiga 520-2192 (Japan); Otani, Hideji, E-mail: otani@belle.shiga-med.ac.jp [Department of Radiology, Shiga University of Medical Science, Setatsukinowa-cho, Otsu, Shiga 520-2192 (Japan); Takahashi, Masashi, E-mail: masashi@belle.shiga-med.ac.jp [Department of Radiology, Shiga University of Medical Science, Setatsukinowa-cho, Otsu, Shiga 520-2192 (Japan); Murata, Kiyoshi, E-mail: murata@belle.shiga-med.ac.jp [Department of Radiology, Shiga University of Medical Science, Setatsukinowa-cho, Otsu, Shiga 520-2192 (Japan); Shiomi, Masashi, E-mail: ieakusm@med.kobe-u.ac.jp [Institute for Experimental Animals, Kobe University School of Medicine, 7-5-1 Kusunoki-cho, Tyuoku, Kobe, Hyogo 650-0017 (Japan); Tabata, Yasuhiko, E-mail: yasuhiko@frontier.kyoto-u.ac.jp [Department of Biomaterials, Institute for Frontier Medical Sciences, Kyoto University, 53 Syogoin-Kawahara-cho, Sakyoku, Kyoto 606-8507 (Japan); Nohara, Satoshi, E-mail: s-nohara@meito-sangyo.co.jp [The Nagoya Research Laboratory, Meito Sangyo Co., Ltd., 25-5 Nishibiwajima-cho, Kiyosu, Aichi 452-0067 (Japan)

2013-11-01

Purpose: We used magnetic resonance imaging (MRI) and histologic techniques to compare the uptake by the rabbit atherosclerotic wall of 4 types of superparamagnetic iron oxide (SPIO) particles, i.e. SPIO, mannan-coated SPIO (M-SPIO), ultrasmall SPIO (USPIO), and mannan-coated USPIO (M-USPIO). Materials and methods: All experimental protocols were approved by our institutional animal experimentation committee. We intravenously injected 12 Watanabe heritable hyperlipidemic rabbits with one of the 4 types of SPIO (0.8 mmol Fe/kg). Two other rabbits served as the control. The rabbits underwent in vivo contrast-enhanced magnetic resonance angiography (MRA) before- and 5 days after these injections; excised aortae were subjected to in vitro MRI. In the in vivo and in vitro studies we assessed the signal intensity of the vessels at identical regions of interest (ROI) and calculated the signal-to-noise ratio (SNR). For histologic assessment we evaluated the iron-positive regions in Prussian blue-stained specimens. Results: There were significant differences in iron-positive regions where M-USPIO > USPIO, M-SPIO > SPIO, USPIO > SPIO (p < 0.05) but not between M-USPIO and M-SPIO. The difference between the pre- and post-injection SNR was significantly greater in rabbits treated with M-USPIO than USPIO and in rabbits injected with M-SPIO than SPIO (p < 0.05). On in vitro MRI scans SNR tended to be lower in M-USPIO- and M-SPIO- than USPIO- and SPIO-treated rabbits (p < 0.1). Conclusion: Histologic and imaging analysis showed that mannan-coated SPIO and USPIO particles were taken up more readily by the atherosclerotic rabbit wall than uncoated SPIO and USPIO.

In vitro removal of toxic heavy metals by poly(γ-glutamic acid-coated superparamagnetic nanoparticles

Directory of Open Access Journals (Sweden)

Inbaraj BS

2012-08-01

Full Text Available Baskaran Stephen Inbaraj,1 Bing-Huei Chen1,21Department of Food Science, 2Graduate Institute of Medicine, Fu Jen University, Taipei, TaiwanBackground: Chelation therapy involving organic chelators for treatment of heavy metal intoxication can cause cardiac arrest, kidney overload, mineral deficiency, and anemia.Methods: In this study, superparamagnetic iron oxide nanoparticles (SPIONs modified with an edible biopolymer poly(γ-glutamic acid (PGA were synthesized by coprecipitation method, characterized and evaluated for their removal efficiency of heavy metals from a metal solution, and simulated gastrointestinal fluid (SGIF.Results: Instrumental characterization of bare- and PGA-SPIONs revealed 7% coating of PGA on SPIONs with a spherical shape and an iron oxide spinel structure belonging to magnetite. The particle sizes as determined from transmission electron microscopy images were 8.5 and 11.7 nm for bare- and PGA-SPIONs, respectively, while the magnetization values were 70.3 and 61.5 emu/g. Upon coating with PGA, the zeta potentials were shifted from positive to negative at most of the environmental pH (3–8 and biological pH (1–8, implying good dispersion in aqueous suspension and favorable conditions for heavy metal removal. Batch studies showed rapid removal of lead and cadmium with the kinetic rates estimated by pseudo-second-order model being 0.212 and 0.424 g/mg•min, respectively. A maximum removal occurred in the pH range 4–8 in deionized water and 5–8 in SGIF corresponding to most gastrointestinal pH except for the stomach. Addition of different ionic strengths (0.001–1 M sodium acetate and essential metals (Cu, Fe, Zn, Mg, Ca, and K did not show any marked influence on lead removal by PGA-SPIONs, but significantly reduced the binding of cadmium. Compared to deionized water, the lead removal from SGIF was high at all pH with the Langmuir monolayer removal capacity being 98.70 mg/g for the former and 147.71 mg/g for the

Subtle cytotoxicity and genotoxicity differences in superparamagnetic iron oxide nanoparticles coated with various functional groups

Directory of Open Access Journals (Sweden)

Hong SC

2011-12-01

Full Text Available Seong Cheol Hong1,*, Jong Ho Lee1,*, Jaewook Lee1, Hyeon Yong Kim1, Jung Youn Park2, Johann Cho3, Jaebeom Lee1, Dong-Wook Han11Department of Nanomedical Engineering, BK21 Nano Fusion Technology Division, College of Nanoscience and Nanotechnology, Pusan National University, 2Department of Biotechnology Research, National Fisheries Research and Development Institute, Busan, 3Electronic Materials Lab, Samsung Corning Precision Materials Co, Ltd, Gumi City, Gyeongsangbukdo, Korea*These authors contributed equally to this workAbstract: Superparamagnetic iron oxide nanoparticles (SPIONs have been widely utilized for the diagnosis and therapy of specific diseases, as magnetic resonance imaging (MRI contrast agents and drug-delivery carriers, due to their easy transportation to targeted areas by an external magnetic field. For such biomedical applications, SPIONs must have multifunctional characteristics, including optimized size and modified surface. However, the biofunctionality and biocompatibility of SPIONs with various surface functional groups of different sizes have yet to be elucidated clearly. Therefore, it is important to carefully monitor the cytotoxicity and genotoxicity of SPIONs that are surfaced-modified with various functional groups of different sizes. In this study, we evaluated SPIONs with diameters of approximately 10 nm and 100~150 nm, containing different surface functional groups. SPIONs were covered with –O-groups, so-called bare SPIONs. Following this, they were modified with three different functional groups – hydroxyl (–OH, carboxylic (–COOH, and amine (–NH2 groups – by coating their surfaces with tetraethyl orthosilicate (TEOS, (3-aminopropyltrimethoxysilane (APTMS, TEOS-APTMS, or citrate, which imparted different surface charges and sizes to the particles. The effects of SPIONs coated with these functional groups on mitochondrial activity, intracellular accumulation of reactive oxygen species, membrane integrity

Creating nanoshell on the surface of titanium hydride bead

Directory of Open Access Journals (Sweden)

PAVLENKO Vyacheslav Ivanovich

2016-12-01

Full Text Available The article presents data on the modification of titanium hydride bead by creating titanium nanoshell on its surface by ion-plasma vacuum magnetron sputtering. To apply titanium nanoshell on the titanium hydride bead vacuum coating plant of multifunctional nanocomposite coatings QVADRA 500 located in the center of high technology was used. Analysis of the micrographs of the original surface of titanium hydride bead showed that the microstructure of the surface is flat, smooth, in addition the analysis of the microstructure of material surface showed the presence of small porosity, roughness, mainly cavities, as well as shallow longitudinal cracks. The presence of oxide film in titanium hydride prevents the free release of hydrogen and fills some micro-cracks on the surface. Differential thermal analysis of both samples was conducted to determine the thermal stability of the initial titanium hydride bead and bead with applied titanium nanoshell. Hydrogen thermal desorption spectra of the samples of the initial titanium hydride bead and bead with applied titanium nanoshell show different thermal stability of compared materials in the temperature range from 550 to 860о C. Titanium nanoshells applied in this way allows increasing the heat resistance of titanium hydride bead – the temperature of starting decomposition is 695о C and temperature when decomposition finishes is more than 1000о C. Modified in this way titanium hydride bead can be used as a filler in the radiation protective materials used in the construction or upgrading biological protection of nuclear power plants.

Decoupling internalization, acidification and phagosomal-endosomal/lysosomal fusion during phagocytosis of InlA coated beads in epithelial cells.

Directory of Open Access Journals (Sweden)

Craig D Blanchette

Full Text Available BACKGROUND: Phagocytosis has been extensively examined in 'professional' phagocytic cells using pH sensitive dyes. However, in many of the previous studies, a separation between the end of internalization, beginning of acidification and completion of phagosomal-endosomal/lysosomal fusion was not clearly established. In addition, very little work has been done to systematically examine phagosomal maturation in 'non-professional' phagocytic cells. Therefore, in this study, we developed a simple method to measure and decouple particle internalization, phagosomal acidification and phagosomal-endosomal/lysosomal fusion in Madin-Darby Canine Kidney (MDCK and Caco-2 epithelial cells. METHODOLOGY/PRINCIPAL FINDINGS: Our method was developed using a pathogen mimetic system consisting of polystyrene beads coated with Internalin A (InlA, a membrane surface protein from Listeria monocytogenes known to trigger receptor-mediated phagocytosis. We were able to independently measure the rates of internalization, phagosomal acidification and phagosomal-endosomal/lysosomal fusion in epithelial cells by combining the InlA-coated beads (InlA-beads with antibody quenching, a pH sensitive dye and an endosomal/lysosomal dye. By performing these independent measurements under identical experimental conditions, we were able to decouple the three processes and establish time scales for each. In a separate set of experiments, we exploited the phagosomal acidification process to demonstrate an additional, real-time method for tracking bead binding, internalization and phagosomal acidification. CONCLUSIONS/SIGNIFICANCE: Using this method, we found that the time scales for internalization, phagosomal acidification and phagosomal-endosomal/lysosomal fusion ranged from 23-32 min, 3-4 min and 74-120 min, respectively, for MDCK and Caco-2 epithelial cells. Both the static and real-time methods developed here are expected to be readily and broadly applicable, as they simply

Mechanisms of complement activation by dextran-coated superparamagnetic iron oxide (SPIO) nanoworms in mouse versus human serum

DEFF Research Database (Denmark)

Banda, Nirmal K; Mehta, Gaurav; Chao, Ying

2014-01-01

BACKGROUND: The complement system is a key component of innate immunity implicated in the neutralization and clearance of invading pathogens. Dextran coated superparamagnetic iron oxide (SPIO) nanoparticle is a promising magnetic resonance imaging (MRI) contrast agent. However, dextran SPIO has...... the mechanisms of human complement activation. Mouse data were analyzed by non-paired t-test, human data were analyzed by ANOVA followed by multiple comparisons with Student-Newman-Keuls test. RESULTS: In mouse sera, SPIO NW triggered the complement activation via the LP, whereas the AP contributes via...... the CP, but that did not affect the total level of C3 deposition on the particles. CONCLUSIONS: There were important differences and similarities in the complement activation by SPIO NW in mouse versus human sera. Understanding the mechanisms of immune recognition of nanoparticles in mouse and human...

Connecting DNA origami structures using the biotin- streptavidin ...

African Journals Online (AJOL)

Aghomotsegin

carbon nanotubes on DNA origami. In order to reconfigure DNA origami pliers, Kuzuya (Kuzuya et al.,. 2011) and colleagues used the strong binding biotin- streptavidin interaction. All these researchers made use of the biotin- streptavidin interaction to functionalize the DNA strand or. DNA origami structures. In this work, we ...

Pharmacokinetics and biodistribution of radiolabeled avidin, streptavidin and biotin

International Nuclear Information System (INIS)

Rosebrough, S.F.

1993-01-01

The extraordinarily high affinity of avidin and streptavidin for biotin may be exploited in a two-step approach for delivering radiolabeled biotin derivatives suitable for imaging and therapy to target-bound streptavidin or avidin conjugated monoclonal antibodies (MAbs). The in vivo pharmacokinetics and biodistribution of radiolabeled avidin, streptavidin (SA) and DTPA-biocytinamide (DTPA-biotin) were studied in the rabbit and dog. SA circulated in the blood similar to other 60 kDa proteins, avidin cleared immediately and DTPA-biotin exhibited plasma clearance by glomerular filtration. (author)

High heat generation ability in AC magnetic field for nano-sized magnetic Y3Fe5O12 powder prepared by bead milling

International Nuclear Information System (INIS)

Aono, Hiromichi; Ebara, Hiroki; Senba, Ryota; Naohara, Takashi; Maehara, Tsunehiro; Hirazawa, Hideyuki; Watanabe, Yuji

2012-01-01

Nano-sized magnetic Y 3 Fe 5 O 12 ferrite having a high heat generation ability in an AC magnetic field was prepared by bead milling. A commercial powder sample (non-milled sample) of ca. 2.9 μm in particle size did not show any temperature enhancement in the AC magnetic field. The heat generation ability in the AC magnetic field improved with a decrease in the average crystallite size for the bead-milled Y 3 Fe 5 O 12 ferrites. The highest heat ability in the AC magnetic field was for the fine Y 3 Fe 5 O 12 powder with a 15-nm crystallite size (the samples were milled for 4 h using 0.1 mmφ beads). The heat generation ability of the excessively milled Y 3 Fe 5 O 12 samples decreased. The main reason for the high heat generation property of the milled samples was ascribed to an increase in the Néel relaxation of the superparamagnetic material. The heat generation ability was not influenced by the concentration of the ferrite powder. For the samples milled for 4 h using 0.1 mmφ beads, the heat generation ability (W g −1 ) was estimated using a 3.58×10 −4 fH 2 frequency (f/kHz) and the magnetic field (H/kA m −1 ), which is the highest reported value of superparamagnetic materials. - Highlights: ► The nano-sized Y 3 Fe 5 O 12 powder prepared by bead-milling has the highest heat generation ability in an AC magnetic field. ► The heat generation properties are ascribed to an increase in the Néel relaxation of the superparamagnetic material. ► The heat ability (W g −1 ) can be estimated using 3.58×10 −4 fH 2 (f=kHz, H=kA m −1 ). ► This is an expectable material for use in a drug delivery system for the thermal coagulation therapy of cancer tumors.

Mesoporous polymer-coated PAN beads for environmental applications. Fabrication, characterisation and uranium adsorption studies

International Nuclear Information System (INIS)

Aly, Z.; Scales, N.; Davis, J.; Lumpkin, G.

2017-01-01

Adsorption of U(VI) and other heavy metals on millimetre sized polymer-coated polyacrylinitrile (PAN) beads was investigated. PAN was used as scaffolds for the polymer layer thus producing porous material of high surface area, improved mechanical strength and improved adsorption capabilities. Extensive U(VI) adsorption studies were undertaken and results modelled using different kinetic and equilibrium models. Parameters including thermodynamic parameters were evaluated. Sorbent capacities were assessed as 124, 16, and 33 mg g"-"1 for PCP, SPP and Dowex at 60 deg C respectively. U(VI) adsorption mechanism for these adsorbents was postulated. Recovered uranium may be used for production of cheap electricity. (author)

Sensitive detection of nucleic acids by PNA hybridization directed co-localization of fluorescent beads

DEFF Research Database (Denmark)

Shiraishi, Takehiko; Deborggraeve, Stijn; Büscher, Philippe

2011-01-01

)avidin-coated fluorescent beads, differing in size and color [green beads (1 µm) and red beads (5.9 µm)], thereby allowing distinct detection of each PNA probe by conventional fluorescence microscopy. These two PNA beads showed easily detectable co-localization when simultaneously hybridizing to a target nucleic acid...

Studying the effect of particle size and coating type on the blood kinetics of superparamagnetic iron oxide nanoparticles.

Science.gov (United States)

Roohi, Farnoosh; Lohrke, Jessica; Ide, Andreas; Schütz, Gunnar; Dassler, Katrin

2012-01-01

Magnetic resonance imaging (MRI), one of the most powerful imaging techniques available, usually requires the use of an on-demand designed contrast agent to fully exploit its potential. The blood kinetics of the contrast agent represent an important factor that needs to be considered depending on the objective of the medical examination. For particulate contrast agents, such as superparamagnetic iron oxide nanoparticles (SPIOs), the key parameters are particle size and characteristics of the coating material. In this study we analyzed the effect of these two properties independently and systematically on the magnetic behavior and blood half-life of SPIOs. Eleven different SPIOs were synthesized for this study. In the first set (a), seven carboxydextran (CDX)-coated SPIOs of different sizes (19-86 nm) were obtained by fractionating a broadly size-distributed CDX-SPIO. The second set (b) contained three SPIOs of identical size (50 nm) that were stabilized with different coating materials, polyacrylic acid (PAA), poly-ethylene glycol, and starch. Furthermore, small PAA-SPIOs (20 nm) were synthesized to gain a global insight into the effects of particle size vs coating characteristics. Saturation magnetization and proton relaxivity were determined to represent the magnetic and imaging properties. The blood half-life was analyzed in rats using MRI, time-domain nuclear magnetic resonance, and inductively coupled plasma optical emission spectrometry. By changing the particle size without modifying any other parameters, the relaxivity r(2) increased with increasing mean particle diameter. However, the blood half-life was shorter for larger particles. The effect of the coating material on magnetic properties was less pronounced, but it had a strong influence on blood kinetics depending on the ionic character of the coating material. In this report we systematically demonstrated that both particle size and coating material influence blood kinetics and magnetic properties of

A solid-phase technique for preparation of no-carrier-added technetium-99m radiopharmaceuticals: application to the streptavidin/biotin system

International Nuclear Information System (INIS)

Dunn-Dufault, Robert; Pollak, Alfred; Fitzgerald, Jane; Thornback, John R.; Ballinger, James R.

2000-01-01

A high effective specific activity (HESA) formulation of a biotin-containing 99m Tc ligand [RP488: dimethyl-Gly-Ser-Cys(Acm)-Lys(Biotin)-Gly] conveniently prepared from solid phase was compared to a typical low effective specific activity (LESA) solution formulation to demonstrate improved targeting to streptavidin in an in vitro assay and in an in vivo rat model. RP488 was coupled to a maleimide-functionalized polyethylene glycol resin via a thiol ether linkage and labeled with 99m Tc-gluconate at room temperature, followed by elution of the HESA 99m Tc-RP488 in saline (minimum specific activity ∼ 1000 TBq/mmol by amino acid analysis). Both HESA and LESA 99m Tc-RP488 labeled at > 90% purity. In vitro, HESA 99m Tc-RP488 incubated with streptavidin-agarose was bound quantitatively, but there was competition from addition of increasing amounts of cold RP488. In rats, radiotracer uptake was evident at the site of implantation of streptavidin-agarose beads for the HESA dose, less uptake of low effective specific activity (LESA) material, and no appreciable uptake in the control rats of the LESA or HESA dose. The target-to-background ratio for HESA 99m Tc-RP488 was 5.4 times that of the control. The solid-phase technology offers a convenient way to prepare high specific activity receptor-targeting 99m Tc radiopharmaceuticals

Enhanced bio-compatibility of ferrofluids of self-assembled superparamagnetic iron oxide-silica core-shell nanoparticles

Digital Repository Service at National Institute of Oceanography (India)

Narayanan, T.N.; Mary, A.P.R.; Swalih, P.K.A.; Kumar, D.S.; Makarov, D.; Albrecht, M.; Puthumana, J.; Anas, A.; Anantharaman, A.

-interacting, monodispersed and hence the synthesis of such nanostructures has great relevance in the realm of nanoscience. Silica-coated superparamagnetic iron oxide nanoparticles based ferrofluids were prepared using polyethylene glycol as carrier fluid by employing a...

Label-acquired magnetorotation for biosensing: An asynchronous rotation assay

International Nuclear Information System (INIS)

Hecht, Ariel; Kinnunen, Paivo; McNaughton, Brandon; Kopelman, Raoul

2011-01-01

This paper presents a novel application of magnetic particles for biosensing, called label-acquired magnetorotation (LAM). This method is based on a combination of the traditional sandwich assay format with the asynchronous magnetic bead rotation (AMBR) method. In label-acquired magnetorotation, an analyte facilitates the binding of a magnetic label bead to a nonmagnetic solid phase sphere, forming a sandwich complex. The sandwich complex is then placed in a rotating magnetic field, where the rotational frequency of the sandwich complex is a function of the amount of analyte attached to the surface of the sphere. Here, we use streptavidin-coated beads and biotin-coated particles as analyte mimics, to be replaced by proteins and other biological targets in future work. We show this sensing method to have a dynamic range of two orders of magnitude.

Preparation and characterization of (3-aminopropyl)triethoxysilane-coated magnetite nanoparticles

International Nuclear Information System (INIS)

Yamaura, M.; Camilo, R.L.; Sampaio, L.C.; Macedo, M.A.; Nakamura, M.; Toma, H.E.

2004-01-01

Magnetite nanoparticles coated with (3-aminopropyl)triethoxysilane, NH 2 (CH 2 ) 3 Si(OC 2 H 5 ) 3 , were prepared by silanization reaction and characterized by X-ray diffractometry, transmission electron microscopy, atomic force microscopy, Fourier transform infrared spectroscopy and magnetization measurements. Both uncoated and organosilane-coated magnetite exhibited superparamagnetic behavior and strong magnetization at room temperature. Basic groups anchored on the external surface of the coated magnetite were observed. The superparamagnetic particles of coated magnetite are able to bind to biological molecules, drugs and metals and in this way remove them from medium by magnetic separation procedures

Acetylcholinesterase immobilized capillary reactors coupled to protein coated magnetic beads: A new tool for plant extract ligand screening

Science.gov (United States)

Vanzolini, Kenia Lourenço; Jiang, Zhengjin; Zhang, Xiaoqi; Vieira, Lucas Campos Curcino; Corrêa, Arlene Gonçalvez; Cardoso, Carmen Lucia; Cass, Quezia Bezerra; Moaddel, Ruin

2013-01-01

The use of immobilized capillary enzyme reactors (ICERs) and enzymes coated to magnetic beads ((NT or CT)-MB) for ligand screening has been adopted as a new technique of high throughput screening (HTS). In this work the selected target was the enzyme acetylcholinesterase (AChE), which acts on the central nervous system and is a validated target for the treatment of Alzheimer’s disease, as well as for new insecticides. A new approach for the screening of plant extracts was developed based on the ligand fishing experiments and zonal chromatography. For that, the magnetic beads were used for the ligand fishing experiments and capillary bioreactors for the activity assays. The latter was employed also under non-linear conditions to determine the affinity constants of known ligands, for the first time, as well as for the active fished ligand. PMID:24148457

SABinder: A Web Service for Predicting Streptavidin-Binding Peptides.

Science.gov (United States)

He, Bifang; Kang, Juanjuan; Ru, Beibei; Ding, Hui; Zhou, Peng; Huang, Jian

2016-01-01

Streptavidin is sometimes used as the intended target to screen phage-displayed combinatorial peptide libraries for streptavidin-binding peptides (SBPs). More often in the biopanning system, however, streptavidin is just a commonly used anchoring molecule that can efficiently capture the biotinylated target. In this case, SBPs creeping into the biopanning results are not desired binders but target-unrelated peptides (TUP). Taking them as intended binders may mislead subsequent studies. Therefore, it is important to find if a peptide is likely to be an SBP when streptavidin is either the intended target or just the anchoring molecule. In this paper, we describe an SVM-based ensemble predictor called SABinder. It is the first predictor for SBP. The model was built with the feature of optimized dipeptide composition. It was observed that 89.20% (MCC = 0.78; AUC = 0.93; permutation test, p binders. In either case, it will be helpful and can benefit related scientific community.

Biotin-Streptavidin Competition Mediates Sensitive Detection of Biomolecules in Enzyme Linked Immunosorbent Assay.

Science.gov (United States)

Lakshmipriya, Thangavel; Gopinath, Subash C B; Tang, Thean-Hock

2016-01-01

Enzyme Linked Immunosorbent Assay (ELISA) is the gold standard assay for detecting and identifying biomolecules using antibodies as the probe. Improving ELISA is crucial for detecting disease-causing agents and facilitating diagnosis at the early stages of disease. Biotinylated antibody and streptavidin-conjugated horse radish peroxide (streptavidin-HRP) often are used with ELISA to enhance the detection of various kinds of targets. In the present study, we used a competition-based strategy in which we pre-mixed free biotin with streptavidin-HRP to generate high-performance system, as free biotin occupies some of the biotin binding sites on streptavidin, thereby providing more chances for streptavidin-HRP to bind with biotinylated antibody. ESAT-6, which is a protein secreted early during tuberculosis infection, was used as the model target. We found that 8 fM of free biotin mixed with streptavidin-HRP anchored the higher detection level of ESAT-6 by four-fold compared with detection without free biotin (only streptavidin-HRP), and the limit of detection of the new method was 250 pM. These results suggest that biotin-streptavidin competition can be used to improve the diagnosis of analytes in other types of sensors.

Monitoring ligand-receptor interactions by photonic force microscopy

Energy Technology Data Exchange (ETDEWEB)

Jeney, Sylvia [M E Mueller Institute for Structural Biology, Biozentrum, University of Basel, Klingelbergstrasse 70, Basel, 4056 (Switzerland); Mor, Flavio; Forro, Laszlo [Laboratory of Complex Matter Physics (LPMC), Ecole Polytechnique Federale de Lausanne (EPFL), CH-1015 Lausanne (Switzerland); Koszali, Roland [Institute for Information and Communication Technologies (IICT), University of Applied Sciences of Western Switzerland (HEIG-VD), Rue Galilee 15, CH 1401 Yverdon-les-bains (Switzerland); Moy, Vincent T, E-mail: sylvia.jeney@unibas.ch, E-mail: vmoy@miami.edu [Department of Physiology and Biophysics, University of Miami Miller School of Medicine, 1600 NW 10th Avenue, Miami, FL 33136 (United States)

2010-06-25

We introduce a method for the acquisition of single molecule force measurements of ligand-receptor interactions using the photonic force microscope (PFM). Biotin-functionalized beads, manipulated with an optical trap, and a streptavidin-functionalized coverslip were used to measure the effect of different pulling forces on the lifetime of individual streptavidin-biotin complexes. By optimizing the design of the optical trap and selection of the appropriate bead size, pulling forces in excess of 50 pN were achieved. Based on the amplitude of three-dimensional (3D) thermal position fluctuations of the attached bead, we were able to select for a bead-coverslip interaction that was mediated by a single streptavidin-biotin complex. Moreover, the developed experimental system was greatly accelerated by automation of data acquisition and analysis. In force-dependent kinetic measurements carried out between streptavidin and biotin, we observed that the streptavidin-biotin complex exhibited properties of a catch bond, with the lifetime increasing tenfold when the pulling force increased from 10 to 20 pN. We also show that silica beads were more appropriate than polystyrene beads for the force measurements, as tethers, longer than 200 nm, could be extracted from polystyrene beads.

Monitoring ligand-receptor interactions by photonic force microscopy

International Nuclear Information System (INIS)

Jeney, Sylvia; Mor, Flavio; Forro, Laszlo; Koszali, Roland; Moy, Vincent T

2010-01-01

We introduce a method for the acquisition of single molecule force measurements of ligand-receptor interactions using the photonic force microscope (PFM). Biotin-functionalized beads, manipulated with an optical trap, and a streptavidin-functionalized coverslip were used to measure the effect of different pulling forces on the lifetime of individual streptavidin-biotin complexes. By optimizing the design of the optical trap and selection of the appropriate bead size, pulling forces in excess of 50 pN were achieved. Based on the amplitude of three-dimensional (3D) thermal position fluctuations of the attached bead, we were able to select for a bead-coverslip interaction that was mediated by a single streptavidin-biotin complex. Moreover, the developed experimental system was greatly accelerated by automation of data acquisition and analysis. In force-dependent kinetic measurements carried out between streptavidin and biotin, we observed that the streptavidin-biotin complex exhibited properties of a catch bond, with the lifetime increasing tenfold when the pulling force increased from 10 to 20 pN. We also show that silica beads were more appropriate than polystyrene beads for the force measurements, as tethers, longer than 200 nm, could be extracted from polystyrene beads.

Secretory production of tetrameric native full-length streptavidin with thermostability using Streptomyces lividans as a host.

Science.gov (United States)

Noda, Shuhei; Matsumoto, Takuya; Tanaka, Tsutomu; Kondo, Akihiko

2015-01-13

Streptavidin is a tetrameric protein derived from Streptomyces avidinii, and has tight and specific biotin binding affinity. Applications of the streptavidin-biotin system have been widely studied. Streptavidin is generally produced using protein expression in Escherichia coli. In the present study, the secretory production of streptavidin was carried out using Streptomyces lividans as a host. In this study, we used the gene encoding native full-length streptavidin, whereas the core region is generally used for streptavidin production in E. coli. Tetrameric streptavidin composed of native full-length streptavidin monomers was successfully secreted in the culture supernatant of S. lividans transformants, and had specific biotin binding affinity as strong as streptavidin produced by E. coli. The amount of Sav using S. lividans was about 9 times higher than using E. coli. Surprisingly, streptavidin produced by S. lividans exhibited affinity to biotin after boiling, despite the fact that tetrameric streptavidin is known to lose its biotin binding ability after brief boiling. We successfully produced a large amount of tetrameric streptavidin as a secretory-form protein with unique thermotolerance.

Water dispersible superparamagnetic Cobalt iron oxide nanoparticles for magnetic fluid hyperthermia

Energy Technology Data Exchange (ETDEWEB)

Salunkhe, Ashwini B. [Centre for advanced materials research, Department of Physics, Savitribai Phule Pune University, Pune 411007 (India); Soft matter and molecular biophysics group, Department of Applied Physics, University of Santiago de Compostela, Santiago de Compostela (Spain); Khot, Vishwajeet M. [Department of Physics and Astronomy, University College London (United Kingdom); Ruso, Juan M. [Soft matter and molecular biophysics group, Department of Applied Physics, University of Santiago de Compostela, Santiago de Compostela (Spain); Patil, S.I., E-mail: patil@physics.unipune.ac.in [Centre for advanced materials research, Department of Physics, Savitribai Phule Pune University, Pune 411007 (India)

2016-12-01

Superparamagnetic nanoparticles of Cobalt iron oxide (CoFe{sub 2}O{sub 4}) are synthesized chemically, and dispersed in an aqueous suspension for hyperthermia therapy application. Different parameters such as magnetic field intensity, particle concentration which regulates the competence of CoFe{sub 2}O{sub 4} nanoparticle as a heating agents in hyperthermia are investigated. Specific absorption rate (SAR) decreases with increase in the particle concentration and increases with increase in applied magnetic field intensity. Highest value of SAR is found to be 91.84 W g{sup −1} for 5 mg. mL{sup −1} concentration. Oleic acid conjugated polyethylene glycol (OA-PEG) coated CoFe{sub 2}O{sub 4} nanoparticles have shown superior cyto-compatibility over uncoated nanoparticles to L929 mice fibroblast cell lines for concentrations below 2 mg. mL{sup −1}. Present work provides the underpinning for the use of CoFe{sub 2}O{sub 4} nanoparticles as a potential heating mediator for magnetic fluid hyperthermia. - Highlights: • Superparamagnetic, water dispersible CoFe{sub 2}O{sub 4} NPs were synthesized by simple and cost effective Co precipitation route. • Effect of coating on various physical and chemical properties of CoFe{sub 2}O{sub 4} NPs were studied. • The effect of coating on induction heating as well as biocompatibility of NPs were studied.

Graphene Oxide Conjugated Magnetic Beads for RNA Extraction.

Science.gov (United States)

Pham, Xuan-Hung; Baek, Ahruem; Kim, Tae Han; Lee, Sang Hun; Rho, Won-Yeop; Chung, Woo-Jae; Kim, Dong-Eun; Jun, Bong-Hyun

2017-08-04

A magnetic material that consists of silica-coated magnetic beads conjugated with graphene oxide (GO) was successfully prepared for facile ribonucleic acid (RNA) extraction. When the GO-modified magnetic beads were applied to separate the RNA from the lysed cell, the cellular RNAs were readily adsorbed to and readily desorbed from the surface of the GO-modified magnetic beads by urea. The amount of RNA extracted by the GO-modified magnetic beads was ≈170 % as much as those of the control extracted by a conventional phenol-based chaotropic solution. These results demonstrate that the facile method of RNA separation by using GO-modified magnetic beads as an adsorbent is an efficient and simple way to purify intact cellular RNAs and/or microRNA from cell lysates. © 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

Identification of RNAIII-binding proteins in Staphylococcus aureus using tethered RNAs and streptavidin aptamers based pull-down assay.

Science.gov (United States)

Zhang, Xu; Zhu, Qing; Tian, Tian; Zhao, Changlong; Zang, Jianye; Xue, Ting; Sun, Baolin

2015-05-15

It has been widely recognized that small RNAs (sRNAs) play important roles in physiology and virulence control in bacteria. In Staphylococcus aureus, many sRNAs have been identified and some of them have been functionally studied. Since it is difficult to identify RNA-binding proteins (RBPs), very little has been known about the RBPs in S. aureus, especially those associated with sRNAs. Here we adopted a tRNA scaffold streptavidin aptamer based pull-down assay to identify RBPs in S. aureus. The tethered RNA was successfully captured by the streptavidin magnetic beads, and proteins binding to RNAIII were isolated and analyzed by mass spectrometry. We have identified 81 proteins, and expressed heterologously 9 of them in Escherichia coli. The binding ability of the recombinant proteins with RNAIII was further analyzed by electrophoresis mobility shift assay, and the result indicates that proteins CshA, RNase J2, Era, Hu, WalR, Pyk, and FtsZ can bind to RNAIII. This study suggests that some proteins can bind to RNA III in S. aureus, and may be involved in RNA III function. And tRSA based pull-down assay is an effective method to search for RBPs in bacteria, which should facilitate the identification and functional study of RBPs in diverse bacterial species.

On-chip magnetic bead-based DNA melting curve analysis using a magnetoresistive sensor

International Nuclear Information System (INIS)

Rizzi, Giovanni; Østerberg, Frederik W.; Henriksen, Anders D.; Dufva, Martin; Hansen, Mikkel F.

2015-01-01

We present real-time measurements of DNA melting curves in a chip-based system that detects the amount of surface-bound magnetic beads using magnetoresistive magnetic field sensors. The sensors detect the difference between the amount of beads bound to the top and bottom sensor branches of the differential sensor geometry. The sensor surfaces are functionalized with wild type (WT) and mutant type (MT) capture probes, differing by a single base insertion (a single nucleotide polymorphism, SNP). Complementary biotinylated targets in suspension couple streptavidin magnetic beads to the sensor surface. The beads are magnetized by the field arising from the bias current passed through the sensors. We demonstrate the first on-chip measurements of the melting of DNA hybrids upon a ramping of the temperature. This overcomes the limitation of using a single washing condition at constant temperature. Moreover, we demonstrate that a single sensor bridge can be used to genotype a SNP. - Highlights: • We apply magnetoresistive sensors to study solid-surface hybridization kinetics of DNA. • We measure DNA melting profiles for perfectly matching DNA duplexes and for a single base mismatch. • We present a procedure to correct for temperature dependencies of the sensor output. • We reliably extract melting temperatures for the DNA hybrids. • We demonstrate direct measurement of differential binding signal for two probes on a single sensor

Studying the effect of particle size and coating type on the blood kinetics of superparamagnetic iron oxide nanoparticles

Directory of Open Access Journals (Sweden)

Roohi F

2012-08-01

Full Text Available Farnoosh Roohi, Jessica Lohrke, Andreas Ide, Gunnar Schütz, Katrin DasslerMR and CT Contrast Media Research, Bayer Pharma AG, Berlin, GermanyPurpose: Magnetic resonance imaging (MRI, one of the most powerful imaging techniques available, usually requires the use of an on-demand designed contrast agent to fully exploit its potential. The blood kinetics of the contrast agent represent an important factor that needs to be considered depending on the objective of the medical examination. For particulate contrast agents, such as superparamagnetic iron oxide nanoparticles (SPIOs, the key parameters are particle size and characteristics of the coating material. In this study we analyzed the effect of these two properties independently and systematically on the magnetic behavior and blood half-life of SPIOs.Methods: Eleven different SPIOs were synthesized for this study. In the first set (a, seven carboxydextran (CDX-coated SPIOs of different sizes (19–86 nm were obtained by fractionating a broadly size-distributed CDX–SPIO. The second set (b contained three SPIOs of identical size (50 nm that were stabilized with different coating materials, polyacrylic acid (PAA, polyethylene glycol, and starch. Furthermore, small PAA–SPIOs (20 nm were synthesized to gain a global insight into the effects of particle size vs coating characteristics. Saturation magnetization and proton relaxivity were determined to represent the magnetic and imaging properties. The blood half-life was analyzed in rats using MRI, time-domain nuclear magnetic resonance, and inductively coupled plasma optical emission spectrometry.Results: By changing the particle size without modifying any other parameters, the relaxivity r2 increased with increasing mean particle diameter. However, the blood half-life was shorter for larger particles. The effect of the coating material on magnetic properties was less pronounced, but it had a strong influence on blood kinetics depending on the

Tethered particle analysis of supercoiled circular DNA using peptide nucleic acid handles

DEFF Research Database (Denmark)

Norregaard, Kamilla; Andersson, Magnus; Nielsen, Peter Eigil

2014-01-01

This protocol describes how to monitor individual naturally supercoiled circular DNA plasmids bound via peptide nucleic acid (PNA) handles between a bead and a surface. The protocol was developed for single-molecule investigation of the dynamics of supercoiled DNA, and it allows the investigation...... of both the dynamics of the molecule itself and of its interactions with a regulatory protein. Two bis-PNA clamps designed to bind with extremely high affinity to predetermined homopurine sequence sites in supercoiled DNA are prepared: one conjugated with digoxigenin for attachment to an anti......-digoxigenin-coated glass cover slide, and one conjugated with biotin for attachment to a submicron-sized streptavidin-coated polystyrene bead. Plasmids are constructed, purified and incubated with the PNA handles. The dynamics of the construct is analyzed by tracking the tethered bead using video microscopy: less...

Francisella tularensis detection using magnetic labels and a magnetic biosensor based on frequency mixing

International Nuclear Information System (INIS)

Meyer, Martin H.F.; Krause, Hans-Joachim; Hartmann, Markus; Miethe, Peter; Oster, Juergen; Keusgen, Michael

2007-01-01

A biosensor that uses resonant coils with a special frequency-mixing technique and magnetic beads as detectable labels has been established for the detection of Francisella tularensis, the causative agent for tularemia. The detection principle is based on a sandwich immunoassay using an anti-Ft antibody for immunofiltration immobilized to ABICAP[reg] polyethylene filters, and biotinylated with streptavidin-coated magnetic beads as labels. The linear detection range of this biosensor was found to be 10 4 -10 6 cfu F. tularensis lipopolysaccharide (LPS) per ml. Tested sample matrices were physiological PBS buffer and rabbit serum

A novel superparamagnetic surface molecularly imprinted nanoparticle adopting dummy template: an efficient solid-phase extraction adsorbent for bisphenol A.

Science.gov (United States)

Lin, Zhenkun; Cheng, Wenjing; Li, Yanyan; Liu, Zhiren; Chen, Xiangping; Huang, Changjiang

2012-03-30

Leakage of the residual template molecules is one of the biggest challenges for application of molecularly imprinted polymer (MIP) in solid-phase extraction (SPE). In this study, bisphenol F (BPF) was adopted as a dummy template to prepare MIP of bisphenol A (BPA) with a superparamagnetic core-shell nanoparticle as the supporter, aiming to avoid residual template leakage and to increase the efficiency of SPE. Characterization and test of the obtained products (called mag-DMIP beads) revealed that these novel nanoparticles not only had excellent magnetic property but also displayed high selectivity to the target molecule BPA. As mag-DMIP beads were adopted as the adsorbents of solid-phase extraction for detecting BPA in real water samples, the recoveries of spiked samples ranged from 84.7% to 93.8% with the limit of detection of 2.50 pg mL(-1), revealing that mag-DMIP beads were efficient SPE adsorbents. Copyright © 2012 Elsevier B.V. All rights reserved.

Conjugating folate on superparamagnetic Fe3O4@Au nanoparticles using click chemistry

International Nuclear Information System (INIS)

Shen, Xiaofang; Ge, Zhaoqiang; Pang, Yuehong

2015-01-01

Gold-coated magnetic core@shell nanoparticles, which exhibit magneto-optical properties, not only enhance the chemical stability of core and biocompatibility of surface, but also provide a combination of multimodal imaging and therapeutics. The conjugation of these tiny nanoparticles with specific biomolecules allows researchers to target the desired location. In this paper, superparamagnetic Fe 3 O 4 @Au nanoparticles were synthesized and functionalized with the azide group on the surface by formation of self-assembled monolayers. Folate (FA) molecules, non-immunogenic target ligands for cancer cells, are conjugated with alkyne and then immobilized on the azide-terminated Fe 3 O 4 @Au nanoparticles through copper(I)-catalyzed azide-alkyne cycloaddition (click reaction). Myelogenous leukemia K562 cells were used as a folate receptor (FR) model, which can be targeted and extracted by magnetic field after interaction with the Fe 3 O 4 @Au–FA nanoparticles. - Graphical abstract: Self-assembled azide-terminated group on superparamagnetic Fe 3 O 4 @Au nanoparticles followed by click reaction with alkyne-functionalized folate, allowing the nanoparticles target folate receptor of cancer cells. - Highlights: • Azidoundecanethiol was coated on the superparamagnetic Fe 3 O 4 @Au nanoparticles by forming self-assembled monolayers. • Alkyne-terminated folate was synthesized from a reaction between the amine and the carboxylic acid. • Conjugation of Fe 3 O 4 @Au nanoparticles with folate was made by copper-catalyzed azide-alkyne cycloaddition click chemistry

Effect of algal density in bead, bead size and bead concentrations ...

African Journals Online (AJOL)

Effect of algal density in bead, bead size and bead concentrations on wastewater nutrient removal. ... African Journal of Biotechnology ... The bioreactor containing algal beads (4 mm diameter) with 1.5 x 106 cells bead-1 (cell stocking) at concentration of 10.66 beads ml-1 wastewater (1:3 bead: wastewater, v/v) achieved ...

Acetylcholinesterase immobilized capillary reactors coupled to protein coated magnetic beads: A new tool for plant extract ligand screening

OpenAIRE

Vanzolini, Kenia Lourenço; Jiang, Zhengjin; Zhang, Xiaoqi; Vieira, Lucas Campos Curcino; Corrêa, Arlene Gonçalvez; Cardoso, Carmen Lucia; Cass, Quezia Bezerra; Moaddel, Ruin

2013-01-01

The use of immobilized capillary enzyme reactors (ICERs) and enzymes coated to magnetic beads ((NT or CT)-MB) for ligand screening has been adopted as a new technique of high throughput screening (HTS). In this work the selected target was the enzyme acetylcholinesterase (AChE), which acts on the central nervous system and is a validated target for the treatment of Alzheimer’s disease, as well as for new insecticides. A new approach for the screening of plant extracts was developed based on t...

Nonlinear Parametric Excitation Effect Induces Stability Transitions in Swimming Direction of Flexible Superparamagnetic Microswimmers.

Science.gov (United States)

Harduf, Yuval; Jin, Dongdong; Or, Yizhar; Zhang, Li

2018-04-05

Microscopic artificial swimmers have recently become highly attractive due to their promising potential for biomedical microrobotic applications. Previous pioneering work has demonstrated the motion of a robotic microswimmer with a flexible chain of superparamagnetic beads, which is actuated by applying an oscillating external magnetic field. Interestingly, they have shown that the microswimmer's orientation undergoes a 90°-transition when the magnetic field's oscillation amplitude is increased above a critical value. This unexpected transition can cause severe problems in steering and manipulation of flexible magnetic microrobotic swimmers. Thus, theoretical understanding and analysis of the physical origins of this effect are of crucial importance. In this work, we investigate this transition both theoretically and experimentally by using numerical simulations and presenting a novel flexible microswimmer with an anisotropic superparamagnetic head. We prove that this effect depends on both frequency and amplitude of the oscillating magnetic field, and demonstrate existence of an optimal amplitude achieving maximal swimming speed. Asymptotic analysis of a minimal two-link model reveals that the changes in the swimmer's direction represent stability transitions, which are induced by a nonlinear parametric excitation.

Effect of endogenous biotin on the applications of streptavidin and biotin in mice

International Nuclear Information System (INIS)

Rusckowski, Mary; Fogarasi, Miklos; Fritz, Benjamin; Hnatowich, Donald J.

1997-01-01

The use of streptavidin-conjugated antibody to pretarget tumors in animals and patients, prior to administration of radiolabeled biotin, has provided encouraging results, in part because of the high affinity of biotin for streptavidin and the rapid whole-body clearance of biotin. However, binding of endogenous biotin to streptavidin may interfere with the clinical potential of this approach. This report evaluates the effect of endogenous biotin on an antibody-streptavidin conjugate in a mouse tumor model. Tumored nude mice were depleted of endogenous biotin by sequential intraperitoneal injections of streptavidin. The assay of serum biotin levels indicated less than 0.5 ng of biotin per mL of serum in treated mice versus 4 ng per mL in untreated animals. Flow cytometric analysis was used on single-cell suspensions of tumor from animals receiving streptavidin-conjugated IgG to detect the presence of the antibody on the cell membrane (with fluoroisothiocyanate-conjugated goat anti-mouse antibody), and to detect biotin binding sites on streptavidin (with biotin-phycoerythrin). Both treated and untreated mice demonstrated the presence of antibody on tumor cells through 48 h postadministration, but only in treated animals were biotin binding sites observed. These results in the mouse model suggest that the small concentration of streptavidin delivered to a tumor via a specific antibody may be saturated with endogenous biotin and therefore not able to be targeted subsequently with radiolabeled biotin

Streptavidin-binding peptides and uses thereof

Science.gov (United States)

Szostak, Jack W. (Inventor); Wilson, David S. (Inventor); Keefe, Anthony D. (Inventor)

2006-01-01

The invention provides peptides with high affinity for streptavidin. These peptides may be expressed as part of fusion proteins to facilitate the detection, quantitation, and purification of proteins of interest.

Artificial Metalloenzymes Based on the Biotin-Streptavidin Technology: Challenges and Opportunities.

Science.gov (United States)

Heinisch, Tillmann; Ward, Thomas R

2016-09-20

The biotin-streptavidin technology offers an attractive means to engineer artificial metalloenzymes (ArMs). Initiated over 50 years ago by Bayer and Wilchek, the biotin-(strept)avidin techonology relies on the exquisite supramolecular affinity of either avidin or streptavidin for biotin. This versatile tool, commonly referred to as "molecular velcro", allows nearly irreversible anchoring of biotinylated probes within a (strept)avidin host protein. Building upon a visionary publication by Whitesides from 1978, several groups have been exploiting this technology to create artificial metalloenzymes. For this purpose, a biotinylated organometallic catalyst is introduced within (strept)avidin to afford a hybrid catalyst that combines features reminiscent of both enzymes and organometallic catalysts. Importantly, ArMs can be optimized by chemogenetic means. Combining a small collection of biotinylated organometallic catalysts with streptavidin mutants allows generation of significant diversity, thus allowing optimization of the catalytic performance of ArMs. Pursuing this strategy, the following reactions have been implemented: hydrogenation, alcohol oxidation, sulfoxidation, dihydroxylation, allylic alkylation, transfer hydrogenation, Suzuki cross-coupling, C-H activation, and metathesis. In this Account, we summarize our efforts in the latter four reactions. X-ray analysis of various ArMs based on the biotin-streptavidin technology reveals the versatility and commensurability of the biotin-binding vestibule to accommodate and interact with transition states of the scrutinized organometallic transformations. In particular, streptavidin residues at positions 112 and 121 recurrently lie in close proximity to the biotinylated metal cofactor. This observation led us to develop a streamlined 24-well plate streptavidin production and screening platform to optimize the performance of ArMs. To date, most of the efforts in the field of ArMs have focused on the use of purified

Enhanced protein adsorption and patterning on nanostructured latex-coated paper.

Science.gov (United States)

Juvonen, Helka; Määttänen, Anni; Ihalainen, Petri; Viitala, Tapani; Sarfraz, Jawad; Peltonen, Jouko

2014-06-01

Specific interactions of extracellular matrix proteins with cells and their adhesion to the substrate are important for cell growth. A nanopatterned latex-coated paper substrate previously shown to be an excellent substrate for cell adhesion and 2D growth was studied for directed immobilization of proteins. The nanostructured latex surface was formed by short-wavelength IR irradiation of a two-component latex coating consisting of a hydrophilic film-forming styrene butadiene acrylonitrile copolymer and hydrophobic polystyrene particles. The hydrophobic regions of the IR-treated latex coating showed strong adhesion of bovine serum albumin (cell repelling protein), fibronectin (cell adhesive protein) and streptavidin. Opposite to the IR-treated surface, fibronectin and streptavidin had a poor affinity toward the untreated pristine latex coating. Detailed characterization of the physicochemical surface properties of the latex-coated substrates revealed that the observed differences in protein affinity were mainly due to the presence or absence of the protein repelling polar and charged surface groups. The protein adsorption was assisted by hydrophobic (dehydration) interactions. Copyright © 2014 Elsevier B.V. All rights reserved.

Synthesis of magnetic hollow silica using polystyrene bead as a template

International Nuclear Information System (INIS)

Wu, W.; Caruntu, D.; Martin, A.; Yu, M.H.; O'Connor, C.J.; Zhou, W.L.; Chen, J.-F.

2007-01-01

In this paper, we report a new route to synthesize novel magnetic hollow silica nanospheres (MHSNs) using polystyrene particles as sacrificial templates, and TEOS and Fe 3 O 4 as precursors. TEM, EDS, XRD, and SQUID were applied to characterize MHSNs. TEM and EDS results show that the MHSNs consist of about 200 nm of hollow cores and ∼35 nm shells with ∼10 nm of Fe 3 O 4 nanoparticles embedded. The polystyrene beads were successfully removed by immersing the as-prepared silica nanocomposite in a toluene solution. XRD results demonstrate that the Fe 3 O 4 magnetic nanoparticles still keep spinel structure even heated at low temperature. The surface status of the polystyrene beads and Fe 3 O 4 nanoparticles has an important effect on the formation of the MHSNs. The MHSNs present a superparamagnetism at room temperature by SQUID measurement. The MHSNs have potential applications in biosystem and nanomedicine

Nanohole 3D-size tailoring through polystyrene bead combustion during thin film deposition

International Nuclear Information System (INIS)

Peng Xiaofeng; Kamiya, Itaru

2009-01-01

A novel approach is presented for nanohole 3D-size tailoring. The process starts with a monolayer of polystyrene (PS) beads spun coat on silicon wafer as a template. The holes can be directly prepared through combustion of PS beads by oxygen plasma during metal or oxide thin film deposition. The incoming particles are prevented from adhering on PS beads by H 2 O and CO 2 generated from the combustion of the PS beads. The hole depth generally depends on the film thickness. The hole diameter can be tailored by the PS bead size, film deposition rate, and also the combustion speed of the PS beads. In this work, a series of holes with depth of 4-24 nm and diameter of 10-36 nm has been successfully prepared. The hole wall materials can be selected from metals such as Au or Pt and oxides such as SiO 2 or Al 2 O 3 . These templates could be suitable for the preparation and characterization of novel nanodevices based on single quantum dots or single molecules, and could be extended to the studies of a wide range of coating materials and substrates with controlled hole depth and diameters.

Silica-supported Macroporous Chitosan Bead for Affinity Purification of Trypsin Inhibitor

Institute of Scientific and Technical Information of China (English)

Feng Na XI; Jian Min WU; Ming Ming LUAN

2005-01-01

Macroporous cross-linking chitosan layer coated on silica gel (CTS-SiO2) was prepared by phase inversion and polyethylene glycol (PEG) molecular imprinting methods. Formation of macroporous surface was investigated by scanning electron microscopy (SEM) and BET analysis.The prepared bead was activated by reacting with 1,2-ethylene diglycidyl ether for introducing epoxy groups, and trypsin could be efficiently immobilized on the bead as a biospecific ligand.The bead bearing trypsin was employed to purify trypsin inhibitor (TIs) from egg white as affinity adsorbent.

Improved tumor localization with (strept)avidin and labeled biotin as a substitute for antibody

International Nuclear Information System (INIS)

Hnatowich, D.J.; Fritz, B.; Virzi, F.; Mardirossian, G.; Rusckowski, M.

1993-01-01

We have investigated tumor localization with labeled biotin administered subsequent to unlabeled and unconjugated streptavidin. Nude mice bearing anti-CEA tumors (LS174T) received 10 μg of 111 In-labeled anti-CEA antibody (C110) or 111 In-labeled streptavidin with sacrifice 5 h later. In an examination of pretargeting, other animals received 50 μg of unlabeled streptavidin followed 3 h later with 1 μg of 111 In-labeled biotin (EB 1 ) and sacrifice 2 h later. The biodistribution of labeled streptavidin was similar to that of labeled specific antibody except for lower blood and higher kidney levels. Tumor levels were also lower with labeled streptavidin but, because of still lower levels in liver and blood, the tumor/normal tissue ratios were improved. When unlabeled streptavidin was administered and followed by labeled biotin (pretargeting), tumor levels were further reduced modestly; however, normal tissue levels were greatly reduced such that the tumor/blood and tumor/liver ratios were 10.6 and 2.2 vs 1.5 and 0.5 for the specific antibody. Improvements were seen in all tissues sampled with the exception of kidney and muscle. A further control of labeled biotin alone (without the preinjection of streptavidin) showed minimal accumulations in all tissues with the exception of kidneys. In conclusion, the accumulation of (strept)avidin by passive diffusion in tumor may be comparable, at early times, to the accumulation of specific antibody. (author)

Immobilization of streptavidin on 4H-SiC for biosensor development

International Nuclear Information System (INIS)

Williams, Elissa H.; Davydov, Albert V.; Motayed, Abhishek; Sundaresan, Siddarth G.; Bocchini, Peter; Richter, Lee J.; Stan, Gheorghe; Steffens, Kristen; Zangmeister, Rebecca; Schreifels, John A.; Rao, Mulpuri V.

2012-01-01

A sequential layer formation chemistry is demonstrated for the functionalization of silicon carbide (SiC) appropriate to biosensing applications. (0 0 0 1) 4H-SiC was functionalized with 3-aminopropyltriethoxysilane (APTES) and subsequently biotinylated for the selective immobilization of streptavidin. Atomic force microscopy, X-ray photoelectron spectroscopy, ellipsometry, fluorescence microscopy, and contact angle measurements were utilized to determine the structure, thickness, wettability, and reactivity of the resulting surface after each functionalization step. Optimization of the APTES layer was found to be critical to the success of the subsequent steps; multilayer, polymeric films resulted in irreproducible behavior. It was shown that there was significant non-specific (electrostatic) binding of streptavidin to APTES functionalized SiC, thus revealing the importance of a uniform biotinylation step prior to streptavidin attachment. The experimental results demonstrate that the APTES functionalized and biotinylated SiC surface has the potential to be employed as a biosensing platform for the selective detection of streptavidin molecules.

Intravenous avidin chase improved localization of radiolabeled streptavidin in intraperitoneal xenograft pretargeted with biotinylated antibody

International Nuclear Information System (INIS)

Zhang Meili; Sakahara, Harumi; Yao Zhengsheng; Saga, Tsuneo; Nakamoto, Yuhi; Sato, Noriko; Nakada, Hiroshi; Yamashina, Ikuo; Konishi, Junji

1997-01-01

In the present study, we examined the effect of avidin administered intravenously (i.v.) on the biodistribution of radiolabeled streptavidin in mice bearing intraperitoneal (IP) xenografts pretargeted with biotinylated antibody. Tumors were established in nude mice by IP inoculation of LS180 human colon cancer cells. Monoclonal antibody MLS128, which recognizes Tn antigen on mucin, was biotinylated and injected IP into the IP tumor-bearing mice. Radioiodinated streptavidin was administered IP or i.v. 48 h after pretargeting of biotinylated antibody. Avidin was administered i.v. 30 min prior to streptavidin injection. The localization of radioiodinated streptavidin in the tumor pretargeted with biotinylated antibody was significantly higher than that without pretargeting and that of radioiodinated MLS128 by the one-step method. Avidin administration significantly accelerated the clearance of radioiodinated streptavidin in blood and other normal tissues and increased the tumor-to-blood radioactivity ratio regardless of administration route of streptavidin. The i.v. avidin chase improved tumor localization of radiolabeled streptavidin in the IP xenografts pretargeted with biotinylated antibody

Francisella tularensis detection using magnetic labels and a magnetic biosensor based on frequency mixing

Energy Technology Data Exchange (ETDEWEB)

Meyer, Martin H.F. [Institute for Pharmaceutical Chemistry, Philipps-Universitaet Marburg (Germany); Krause, Hans-Joachim [Institute of Bio-and Nanosystems (IBN-2), Research Center Juelich (Germany); Hartmann, Markus [Institute for Pharmaceutical Chemistry, Philipps-Universitaet Marburg (Germany); Miethe, Peter [SENOVA GmbH, Jena (Germany); Oster, Juergen [chemagen GmbH, Baesweiler (Germany); Keusgen, Michael [Institute for Pharmaceutical Chemistry, Philipps-Universitaet Marburg (Germany)]. E-mail: Keusgen@staff.uni-marburg.de

2007-04-15

A biosensor that uses resonant coils with a special frequency-mixing technique and magnetic beads as detectable labels has been established for the detection of Francisella tularensis, the causative agent for tularemia. The detection principle is based on a sandwich immunoassay using an anti-Ft antibody for immunofiltration immobilized to ABICAP[reg] polyethylene filters, and biotinylated with streptavidin-coated magnetic beads as labels. The linear detection range of this biosensor was found to be 10{sup 4}-10{sup 6} cfu F. tularensis lipopolysaccharide (LPS) per ml. Tested sample matrices were physiological PBS buffer and rabbit serum.

The Motion of a Single Molecule, the Lambda-Receptor, in the Bacterial Outer Membrane

DEFF Research Database (Denmark)

Oddershede, Lene; Dreyer, Jakob Kisbye; Grego, Sonia

2002-01-01

Using optical tweezers and single particle tracking, we have revealed the motion of a single protein, the lambda-receptor, in the outer membrane of living Escherichia coli bacteria. We genetically modified the lambda-receptor placing a biotin on an extracellular site of the receptor in vivo....... The efficiency of this in vivo biotinylation is very low, thus enabling the attachment of a streptavidin-coated bead binding specifically to a single biotinylated lambda-receptor. The bead was used as a handle for the optical tweezers and as a marker for the single particle tracking routine. We propose a model...

Synthesis, characterization and antibacterial activity of superparamagnetic nanoparticles modified with glycol chitosan

International Nuclear Information System (INIS)

Stephen Inbaraj, Baskaran; Tsai, Tsung-Yu; Chen, Bing-Huei

2012-01-01

Iron oxide nanoparticles (IONPs) were synthesized by coprecipitation of iron salts in alkali media followed by coating with glycol chitosan (GC-coated IONPs). Both bare and GC-coated IONPs were subsequently characterized and evaluated for their antibacterial activity. Comparison of Fourier transform infrared spectra and thermogravimetric data of bare and GC-coated IONPs confirmed the presence of GC coating on IONPs. Magnetization curves showed that both bare and GC-coated IONPs are superparamagnetic and have saturation magnetizations of 70.3 and 59.8 emu g −1 , respectively. The IONP size was measured as ∼8–9 nm by transmission electron microscopy, and their crystal structure was assigned to magnetite from x-ray diffraction patterns. Both bare and GC-coated IONPs inhibited the growths of Escherichia coli ATCC 8739 and Salmonella enteritidis SE 01 bacteria better than the antibiotics linezolid and cefaclor, as evaluated by the agar dilution assay. GC-coated IONPs showed higher potency against E. coli O157:H7 and Staphylococcus aureus ATCC 10832 than bare IONPs. Given their biocompatibility and antibacterial properties, GC-coated IONPs are a potential nanomaterial for in vivo applications.

Floating-bioadhesive gastroretentive Caesalpinia pulcherrima-based beads of amoxicillin trihydrate for Helicobacter pylori eradication.

Science.gov (United States)

Thombre, Nilima A; Gide, Paraag S

2016-01-01

An oral dosage form containing floating bioadhesive gastroretentive microspheres forms a stomach-specific drug delivery system for the treatment of Helicobacter pylori. To prepare and evaluate controlled release floating bioadhesive gastroretentive chitosan-coated amoxicillin trihydrate-loaded Caesalpinia pulcherrima galactomannan (CPG)-alginate beads (CCA-CPG-A), for H. pylori eradication. CCA-CPG-A beads were prepared by ionotropic gelation, using 2(3) factorial design with quantity of drug, combination of CPG with sodium alginate and concentration of calcium chloride as variables. Beads facilitated mucoadhesion to gastric mucosa with floating nature caused by chitosan coating for wide distribution throughout GIT. Developed beads were evaluated for characteristics like beads size-morphology, entrapment efficiency, DSC, XRD, FTIR, swelling ratio, in vitro mucoadhesion, in vitro drug release, in vitro floating and in vitro H. pylori growth inhibition studies. CCA-CPG-A beads were studied in Wistar rats for in vivo gastric mucoadhesion, in vivo H. pylori growth inhibition studies using PCR amplification of isolated DNA, rapid urease test. Developed beads possess drug release of 79-92%, entrapment efficiency of 65-89%, mucoadhesion of 61-89%. In vivo mucoadhesion study showed more than 85% mucoadhesion of beads even after 7th hour. In vitro-in vivo growth inhibition study showed complete eradication of H. pylori. CPG-alginate and chitosan in beads interacts with gastric mucosubstrate surface for prolonged gastric residence with floating bioadhesion mechanism for H. pylori eradication in rats. Floating bioadhesive CCA-CPG-A beads offer a promising drug delivery system for H. pylori eradication at lower dose, reduced adverse effect and enhance bioavailability.

Improved tumor localization with (strept)avidin and labeled biotin as a substitute for antibody

International Nuclear Information System (INIS)

Hnatowich, D.J.; Fritz, B.; Virzi, F.; Mardirossian, G.; Rusckowski, M.

1993-01-01

We have investigated tumor localization with labeled biotin administered subsequent to unlabeled and unconjugated streptavidin. Nude mice bearing anti-CEA tumors (LS174T) received 10 μg of 111 In-labeled anti-CEA antibody (C110) or 111 In-labeled streptavidin with sacrifice 5 h later. In an examination of pretargeting, other animals received 50 μg of unlabeled streptavidin followed 3 h later with 1 μg of 111 In-labeled biotin (EB 1 ) and sacrifice 2 h later. The biodistribution of labeled streptavidin was similar to that of labeled specific antibody except for lower blood and higher kidney levels. Tumor levels were also lower with labeled streptavidin but, because of still lower levels in liver and blood, the tumor/normal tissue ratios were improved. When unlabeled streptavidin was administered and followed by labeled biotin (pretargeting), tumor levels were further reduced modestly; however, normal tissue levels were greatly reduced such that the tumor/blood and tumor/liver ratios were 10.6 and 2.2 vs 1.5 and 0.5 for the specific antibody. Improvements were seen in all tissues sampled with the exception of kidney and muscle. A further control of labeled biotin alone showed minimal accumulation in all tissues with the exception of kidneys. In conclusion, the accumulation of (strept)avidin by passive diffusion in tumor may be comparable, at early times, to the accumulation of specific antibody. By combining the administration of unlabeled (strept)avidin with labeled biotin, tumor targeting may potentially be improved. (author)

Effect of algal density in bead, bead size and bead concentrations ...

African Journals Online (AJOL)

Laboratory experiments were performed to study nitrogen and phosphorus uptake by the unicellular green microalga Chlorella vulgaris immobilized in calcium alginate beads. Different cell stockings in beads, different bead sizes and different algal bead concentrations in wastewaters were tested. Significant higher nutrients ...

Fast removal of malachite green dye using novel superparamagnetic sodium alginate-coated Fe3O4 nanoparticles.

Science.gov (United States)

Mohammadi, Abbas; Daemi, Hamed; Barikani, Mehdi

2014-08-01

In this study, superparamagnetic sodium alginate-coated Fe3O4 nanoparticles (Alg-Fe3O4) as a novel magnetic adsorbent were prepared by in situ coprecipitation method, in which Fe3O4 nanoparticles were precipitated from FeCl3 and FeCl2 under alkaline medium in the presence of sodium alginate. The Alg-Fe3O4 nanoparticles were used for removal of malachite green (MG) from aqueous solutions using batch adsorption technique. The characterization of synthesized nanoparticles was performed using XRD, FTIR, TEM, TGA and vibrating sample magnetometer (VSM) techniques. FTIR analysis of synthesized nanoparticles provided the evidence that sodium alginate was successfully coated on the surface of Fe3O4 nanoparticles. The FT-IR and TGA characterization showed that the Alg-Fe3O4 nanoparticles contained about 14% (w/w) of sodium alginate. Moreover, TEM analysis indicated that the average diameter of the Alg-Fe3O4 nanoparticles was about 12nm. The effects of adsorbent dosage, pH and temperature were investigated on the adsorption properties of MG onto Alg-Fe3O4 nanoparticles. The equilibrium adsorption data were modeled using the Langmuir and Freundlich isotherms. The maximum adsorption capacity obtained from Langmuir isotherm equation was 47.84mg/g. The kinetics of adsorption of MG onto Alg-Fe3O4 nanoparticles were investigated using the pseudo-first-order and pseudo-second-order kinetic models. The results showed that the adsorption of MG onto nanoparticles followed pseudo-second-order kinetic model. Copyright © 2014 Elsevier B.V. All rights reserved.

The Role of an Aggrecan 32mer Fragment in Post-Traumatic Osteoarthritis

Science.gov (United States)

2017-10-01

streptavidin-coated AlphaLISA donor beads and incubate for 30 minutes.  Read sample absorbance at 615nm on the PerkinElmer Enspire plate reader. 11...HRPO approval within the next six months. Nothing to report 15 Significant changes in use or care of human subjects, vertebrate animals ...human subjects, vertebrate animals , biohazards, and/or select agents during the reporting period. If required, were these changes approved by the

A half-ring GMR sensor for detection of magnetic beads immobilized on a circular micro-trap

KAUST Repository

Gooneratne, Chinthaka Pasan

2011-11-01

Utilizing magnetic principles in biological immunoassays is an attractive option given its ability to remotely and non-invasively manipulate and detect cells tagged with micro/nano size superparamagnetic type beads and due to the fact that even the most complex biological immunoassays will have very little magnetic effect. The presence of magnetic beads can be detected by a magnetic sensor which quantifies the amount of target cells present in the immunoassay. In order to increase the detection rate a circular conducting micro-trap is employed to attract, trap and transport the magnetic beads to the sensing area. In this research we propose a half-ring spin valve type giant magnetoresistance (GMR) sensor for the measurement of stray fields produced by 2 μm magnetic beads which are around the circular micro-trap. A couple of half-ring GMR sensors can be used to cover the entire circular border width, in order to detect the majority of the immobilized magnetic beads. Analytical and numerical analysis leading towards the fabrication of the half-ring GMR sensor are presented. DC characterization of the fabricated sensor showed a magnetoresistance of 5.9 %. Experimental results showed that the half-ring GMR sensor detected the presence of 2 μm magnetic beads. Hence, half-ring GMR sensors integrated with a circular micro-trap have great potential to be used as an effective disease diagnostic device. © 2011 IEEE.

A half-ring GMR sensor for detection of magnetic beads immobilized on a circular micro-trap

KAUST Repository

Gooneratne, Chinthaka Pasan; Liang, Cai; Useinov, Arthur; Kosel, Jü rgen; Giouroudi, Ioanna

2011-01-01

Utilizing magnetic principles in biological immunoassays is an attractive option given its ability to remotely and non-invasively manipulate and detect cells tagged with micro/nano size superparamagnetic type beads and due to the fact that even the most complex biological immunoassays will have very little magnetic effect. The presence of magnetic beads can be detected by a magnetic sensor which quantifies the amount of target cells present in the immunoassay. In order to increase the detection rate a circular conducting micro-trap is employed to attract, trap and transport the magnetic beads to the sensing area. In this research we propose a half-ring spin valve type giant magnetoresistance (GMR) sensor for the measurement of stray fields produced by 2 μm magnetic beads which are around the circular micro-trap. A couple of half-ring GMR sensors can be used to cover the entire circular border width, in order to detect the majority of the immobilized magnetic beads. Analytical and numerical analysis leading towards the fabrication of the half-ring GMR sensor are presented. DC characterization of the fabricated sensor showed a magnetoresistance of 5.9 %. Experimental results showed that the half-ring GMR sensor detected the presence of 2 μm magnetic beads. Hence, half-ring GMR sensors integrated with a circular micro-trap have great potential to be used as an effective disease diagnostic device. © 2011 IEEE.

Synthesis and characterization of superparamagnetic polymeric nanocapsules

Energy Technology Data Exchange (ETDEWEB)

Grillo, Renato; Fraceto, Leonardo Fernandes, E-mail: renato.grillo@ymail.com [Universidade Estadual Paulista Julio de Mesquita Filho (UNESP), Sorocaba, SP (Brazil); Gallo, Juan; Grando Stroppa, Daniel; Carbo-Argibay, Enrique; Banõbre-Lopez, Manuel [International Iberian Nanotechnology Laboratory, Braga (Portugal); Lima, Renata de [Universidade de Sorocaba (UNISO), SP (Brazil)

2016-07-01

Full text: A wide variety of applications have been considered for superparamagnetic iron oxide nanoparticles (SPIONs), such as magnetic resonance imaging, cancer therapy and remediation of contaminants [1].Polymeric nanostructures (PNS) have also received great interest as suitable encapsulating agents and carriers due to their ability to influence the delivery profile. Hybrid nanosystems have been explored as a synergic approach that combines the modified active release induced by the polymer encapsulation and the intrinsic properties from the inorganic nanoparticles [2]. In this context, poly-ε-caprolactone nanocapsules containing different concentration of ∼8 nm superparamagnetic oleic acid coated magnetite (Fe{sub 3}O{sub 4}@OA) nanoparticles were developed. Successful incorporation of the magnetic nanoparticles was confirmed by transmission electron microscopy coupled with energy dispersive X-ray (TEM-EDX). Results showed that they accumulate preferentially in the outer organic membrane of the PNS. On the other hand, scanning electron microscopy and dynamic light scattering measurements showed a significant increase in particle size from ca. 400 to 800 nm. Magnetic measurements as a function of the applied magnetic field and temperature were performed in both vibrant sample (VSM) and superconducting quantum interference device magnetometers (SQUID). Hysteresis loops showed a superparamagnetic behavior with increasing saturation magnetization as magnetite concentration was progressively incorporated into the PNS. Zero-field cooled and field-cooled (ZFC-FC) magnetic curves showed a shift of the blocking temperature to higher temperatures as the content of magnetite increases in the capsules. These results are promising and contribute to a better understanding of the interaction between magnetic nanoparticles and PNS. References: [1] L. Zhang, W. Dong, H. Sun. Nanoscale 5, 7664-7684 (2013) [2] K.T. Nguyen and Y.L. Zhao. Acc. Chem. Res. 48, 3016-3025 (2015

Magnetic nanoparticle-loaded alginate beads for local micro-actuation of in vitro tissue constructs.

Science.gov (United States)

Alshehri, Awatef M; Wilson, Otto C; Dahal, Bishnu; Philip, John; Luo, Xiaolong; Raub, Christopher B

2017-11-01

Magnetic nanoparticles (MNPs) self-align and transduce magnetic force, two properties which lead to promising applications in cell and tissue engineering. However, the toxicity of MNPs to cells which uptake them is a major impediment to applications in engineered tissue constructs. To address this problem, MNPs were embedded in millimeter-scale alginate beads, coated with glutaraldehyde cross-linked chitosan, and loaded in acellular and MDA-MB-231 cancer cell-seeded collagen hydrogels, providing local micro-actuation under an external magnetic field. Brightfield microscopy was used to assess nanoparticle diffusion from the bead. Phase contrast microscopy and digital image correlation were used to track collagen matrix displacement and estimate intratissue strain under magnetic actuation. Coating the magnetic alginate beads with glutaraldehyde-chitosan prevents bulk diffusion of nanoparticles into the surrounding microenvironment. Further, the beads exert force on the surrounding collagen gel and cells, resulting in intratissue strains of 0-10% tunable with bead dimensions, collagen density, and distance from the bead. Cells seeded adjacent to the embedded beads are subjected to strain gradients without loss of cell viability over two days culture. This study describes a simple way to fabricate crosslinked magnetic alginate beads to load in a collagen tissue construct without direct exposure of the construct to nanoparticles. The findings are significant to in vitro studies of mechanobiology in enabling precise control over dynamic mechanical loading of tissue constructs. Copyright © 2017 Elsevier B.V. All rights reserved.

Novel biosensor system model based on fluorescence quenching by a fluorescent streptavidin and carbazole-labeled biotin.

Science.gov (United States)

Zhu, Xianwei; Shinohara, Hiroaki; Miyatake, Ryuta; Hohsaka, Takahiro

2016-10-01

In the present study, a novel molecular biosensor system model was designed by using a couple of the fluorescent unnatural mutant streptavidin and the carbazole-labeled biotin. BODIPY-FL-aminophenylalanine (BFLAF), a fluorescent unnatural amino acid was position-specifically incorporated into Trp120 position of streptavidin by four-base codon method. On the other hand, carbazole-labeled biotin was synthesized as a quencher for the fluorescent Trp120BFLAF mutant streptavidin. The fluorescence of fluorescent Trp120BFLAF mutant streptavidin was decreased as we expected when carbazole-labeled biotin was added into the mutant streptavidin solution. Furthermore, the fluorescence decrease of Trp120BFLAF mutant streptavidin with carbazole-labeled biotin (100 nM) was recovered by the competitive addition of natural biotin. This result demonstrated that by measuring the fluorescence quenching and recovery, a couple of the fluorescent Trp120BFLAF mutant streptavidin and the carbazole-labeled biotin were successfully applicable for quantification of free biotin as a molecular biosensor system. Copyright © 2016 John Wiley & Sons, Ltd. Copyright © 2016 John Wiley & Sons, Ltd.

Sampling polyhexamethylene guanidine aerosols using eosin Y-coated glass beads

International Nuclear Information System (INIS)

Choi, Sang Hyun; Park, Seon Kyung; Kang, Hyun Joong; Kwon, Jung Hwan; Lee, Kyu Hong

2015-01-01

Fatalities caused by the use of polyhexamethylene guanidine (PHMG), a general-purpose chemical germicide used as a humidifier disinfectant in Korea, have raised concerns about exposure to biocide aerosols in indoor environments. A sampler capable of accumulating PHMG from aqueous aerosols was developed as an alternative to low-volume air samplers. This sampler was prepared by placing glass beads coated with 2-(2,4,5,7-tetrabromo-6-oxido-3-oxo-3H-xanthen-9-yl)benzoate (Eosin Y) in a custom-made plastic holder. Passive sampling rates, measured in a bench-top exposure chamber at two different aqueous PHMG aerosol generation rates, were found to be independent of the experimental conditions. This suggests that the capacity of the sampler to accumulate the PHMG aerosol was sufficient for the sampling duration tested. However, the passive sampling rate was 7.6 × 10"−"6 m"3/h for the sampler area of 22 cm"2. This rate is lower than the typical human breathing rate and inadequate for quantitative instrumental analyses at low concentrations in indoor air. A 30-fold enhancement of the sampling rate was achieved by forced convection using a commercial battery-operated fan at ≥2000 rpm. With this accelerated sampling rate, the sampler could be used to monitor time-integrated concentrations of PHMG aerosols in the air

Sampling polyhexamethylene guanidine aerosols using eosin Y-coated glass beads

Energy Technology Data Exchange (ETDEWEB)

Choi, Sang Hyun; Park, Seon Kyung; Kang, Hyun Joong; Kwon, Jung Hwan [Korea University, Seoul (Korea, Republic of); Lee, Kyu Hong [Korea Institute of Toxicology, Jeongeup (Korea, Republic of)

2015-07-15

Fatalities caused by the use of polyhexamethylene guanidine (PHMG), a general-purpose chemical germicide used as a humidifier disinfectant in Korea, have raised concerns about exposure to biocide aerosols in indoor environments. A sampler capable of accumulating PHMG from aqueous aerosols was developed as an alternative to low-volume air samplers. This sampler was prepared by placing glass beads coated with 2-(2,4,5,7-tetrabromo-6-oxido-3-oxo-3H-xanthen-9-yl)benzoate (Eosin Y) in a custom-made plastic holder. Passive sampling rates, measured in a bench-top exposure chamber at two different aqueous PHMG aerosol generation rates, were found to be independent of the experimental conditions. This suggests that the capacity of the sampler to accumulate the PHMG aerosol was sufficient for the sampling duration tested. However, the passive sampling rate was 7.6 × 10{sup −6} m{sup 3}/h for the sampler area of 22 cm{sup 2}. This rate is lower than the typical human breathing rate and inadequate for quantitative instrumental analyses at low concentrations in indoor air. A 30-fold enhancement of the sampling rate was achieved by forced convection using a commercial battery-operated fan at ≥2000 rpm. With this accelerated sampling rate, the sampler could be used to monitor time-integrated concentrations of PHMG aerosols in the air.

Enhanced pulsed magneto-motive ultrasound imaging using superparamagnetic nanoclusters

International Nuclear Information System (INIS)

Mehrmohammadi, M; Qu, M; Emelianov, S Y; Yoon, K Y; Johnston, K P

2011-01-01

Recently, pulsed magneto-motive ultrasound (pMMUS) imaging augmented with ultra-small magnetic nanoparticles has been introduced as a tool capable of imaging events at molecular and cellular levels. The sensitivity of a pMMUS system depends on several parameters, including the size, geometry and magnetic properties of the nanoparticles. Under the same magnetic field, larger magnetic nanostructures experience a stronger magnetic force and produce larger displacement, thus improving the sensitivity and signal-to-noise ratio (SNR) of pMMUS imaging. Unfortunately, large magnetic iron-oxide nanoparticles are typically ferromagnetic and thus are very difficult to stabilize against colloidal aggregation. In the current study we demonstrate improvement of pMMUS image quality by using large size superparamagnetic nanoclusters characterized by strong magnetization per particle. Water-soluble magnetic nanoclusters of two sizes (15 and 55 nm average size) were synthesized from 3 nm iron precursors in the presence of citrate capping ligand. The size distribution of synthesized nanoclusters and individual nanoparticles was characterized using dynamic light scattering (DLS) analysis and transmission electron microscopy (TEM). Tissue mimicking phantoms containing single nanoparticles and two sizes of nanoclusters were imaged using a custom-built pMMUS imaging system. While the magnetic properties of citrate-coated nanoclusters are identical to those of superparamagnetic nanoparticles, the magneto-motive signal detected from nanoclusters is larger, i.e. the same magnetic field produced larger magnetically induced displacement. Therefore, our study demonstrates that clusters of superparamagnetic nanoparticles result in pMMUS images with higher contrast and SNR.

Magnetic polymeric microspheres for protein adsorption

International Nuclear Information System (INIS)

Felinto, M.C.F.C.; Parra, D.F.; Lugao, A.B.; Batista, M.P.; Higa, O.Z.; Yamaura, M.; Camilo, R.L.; Ribela, M.T.C.P.; Sampaio, L.C.

2005-01-01

Magnetic beads consisting of polymer-coated manganese ferrite nanoparticles were prepared by the precipitation reaction of manganese ferrite into the channels of methyl methacrylate polymer beads by sodium hydroxide, resulting in MnMagBead. MnMagBead was characterized by infrared spectra (FTIR), thermogravimetric analysis of TGA/DTG and indicates the presence of -CO (carbonyl) groups and the MnFe 2 O 4 on the beads. Magnetization measurements were obtained at room temperature in magnetic fields up to 10 KOe using a vibrating sample magnetometer. Introductory Protein adsorption biological tests were processed using labeled I-125 albumin (BSA), and the activity was measured in a gamma counting spectrometer. These superparamagnetic beads exhibit the capacity to bind biological molecules such as proteins like albumin, with a good capability (5 x 10 -6 ) μg/100 mg of beads as compared with other magnetic resins studied in our group

Electrical control of superparamagnetism

Science.gov (United States)

Yamada, Kihiro T.; Koyama, Tomohiro; Kakizakai, Haruka; Miwa, Kazumoto; Ando, Fuyuki; Ishibashi, Mio; Kim, Kab-Jin; Moriyama, Takahiro; Ono, Shimpei; Chiba, Daichi; Ono, Teruo

2017-01-01

The electric field control of superparamagnetism is realized using a Cu/Ni system, in which the deposited Ni shows superparamagnetic behavior above the blocking temperature. An electric double-layer capacitor (EDLC) with the Cu/Ni electrode and a nonmagnetic counter electrode is fabricated to examine the electric field effect on magnetism in the magnetic electrode. By changing the voltage applied to the EDLC, the blocking temperature of the system is clearly modulated.

TiO₂ beads and TiO₂-chitosan beads for urease immobilization.

Science.gov (United States)

Ispirli Doğaç, Yasemin; Deveci, Ilyas; Teke, Mustafa; Mercimek, Bedrettin

2014-09-01

The aim of the present study is to synthesize TiO2 beads for urease immobilization. Two different strategies were used to immobilize the urease on TiO2 beads. In the first method (A), urease enzyme was immobilized onto TiO2 beads by adsorption and then crosslinking. In the second method (B), TiO2 beads were coated with chitosan-urease mixture. To determine optimum conditions of immobilization, different parameters were investigated. The parameters of optimization were initial enzyme concentration (0.5; 1; 1.5; 2mg/ml), alginate concentration (1; 2; 3%), glutaraldehyde concentration (1; 2; 3% v/v) and chitosan concentration (2; 3; 4 mg/ml). The optimum enzyme concentrations were determined as 1.5mg/ml for A and 1.0mg/ml for B. The other optimum conditions were found 2.0% (w/v) for alginate concentration (both A and B); 3.0mg/ml for chitosan concentration (B) and 2.0% (v/v) for glutaraldehyde concentration (A). The optimum temperature (20-60°C), optimum pH (3.0-10.0), kinetic parameters, thermal stability (4-70°C), pH stability (4.0-9.0), operational stability (0-230 min) and reusability (20 times) were investigated for characterization. The optimum temperatures were 30°C (A), 40°C (B) and 35°C (soluble). The temperature profiles of the immobilized ureases were spread over a large area. The optimum pH values for the soluble urease and immobilized urease prepared by using methods (A) and (B) were found to be 7.5, 7.0, 7.0, respectively. The thermal stabilities of immobilized enzyme sets were studied and they maintained 50% activity at 65°C. However, at this temperature free urease protected only 15% activity. Copyright © 2014 Elsevier B.V. All rights reserved.

Labeling transplanted mice islet with polyvinylpyrrolidone coated superparamagnetic iron oxide nanoparticles for in vivo detection by magnetic resonance imaging

Energy Technology Data Exchange (ETDEWEB)

Huang Hai; Xie Qiuping; Kang Muxing; Zhang Bo; Wu Yulian [Department of Surgery, 2nd Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou 310009 (China); Zhang Hui; Chen Jin; Zhai Chuanxin; Yang Deren [State Key Lab of Silicon Materials and Department of Materials Science and Engineering, Zhejiang University, Hangzhou 310027 (China); Jiang Biao, E-mail: wuyulian@medmail.com.c, E-mail: yulianwu2003@yahoo.c [Department of Radiology, 2nd Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou 310009 (China)

2009-09-09

Superparamagnetic iron oxide nanoparticles (SPIO) are emerging as a novel probe for noninvasive cell tracking with magnetic resonance imaging (MRI) and have potential wide usage in medical research. In this study, we have developed a method using high-temperature hydrolysis of chelate metal alkoxide complexes to synthesize polyvinylpyrrolidone coated iron oxide nanoparticles (PVP-SPIO), as a biocompatible magnetic agent that can efficiently label mice islet {beta}-cells. The size, crystal structure and magnetic properties of the as-synthesized nanoparticles have been characterized. The newly synthesized PVP-SPIO with high stability, crystallinity and saturation magnetization can be efficiently internalized into {beta}-cells, without affecting viability and function. The imaging of 100 PVP-SPIO-labeled mice islets in the syngeneic renal subcapsular model of transplantation under a clinical 3.0 T MR imager showed high spatial resolution in vivo. These results indicated the great potential application of the PVP-SPIO as an MRI contrast agent for monitoring transplanted islet grafts in the clinical management of diabetes in the near future.

Labeling transplanted mice islet with polyvinylpyrrolidone coated superparamagnetic iron oxide nanoparticles for in vivo detection by magnetic resonance imaging

International Nuclear Information System (INIS)

Huang Hai; Xie Qiuping; Kang Muxing; Zhang Bo; Wu Yulian; Zhang Hui; Chen Jin; Zhai Chuanxin; Yang Deren; Jiang Biao

2009-01-01

Superparamagnetic iron oxide nanoparticles (SPIO) are emerging as a novel probe for noninvasive cell tracking with magnetic resonance imaging (MRI) and have potential wide usage in medical research. In this study, we have developed a method using high-temperature hydrolysis of chelate metal alkoxide complexes to synthesize polyvinylpyrrolidone coated iron oxide nanoparticles (PVP-SPIO), as a biocompatible magnetic agent that can efficiently label mice islet β-cells. The size, crystal structure and magnetic properties of the as-synthesized nanoparticles have been characterized. The newly synthesized PVP-SPIO with high stability, crystallinity and saturation magnetization can be efficiently internalized into β-cells, without affecting viability and function. The imaging of 100 PVP-SPIO-labeled mice islets in the syngeneic renal subcapsular model of transplantation under a clinical 3.0 T MR imager showed high spatial resolution in vivo. These results indicated the great potential application of the PVP-SPIO as an MRI contrast agent for monitoring transplanted islet grafts in the clinical management of diabetes in the near future.

Magnetic tweezers with high permeability electromagnets for fast actuation of magnetic beads

Energy Technology Data Exchange (ETDEWEB)

Chen, La; Offenhäusser, Andreas; Krause, Hans-Joachim [Institute of Bioelectronics (ICS-8/PGI-8), Forschungszentrum Jülich GmbH, 52425 Jülich (Germany)

2015-04-15

As a powerful and versatile scientific instrument, magnetic tweezers have been widely used in biophysical research areas, such as mechanical cell properties and single molecule manipulation. If one wants to steer bead position, the nonlinearity of magnetic properties and the strong position dependence of the magnetic field in most magnetic tweezers lead to quite a challenge in their control. In this article, we report multi-pole electromagnetic tweezers with high permeability cores yielding high force output, good maneuverability, and flexible design. For modeling, we adopted a piece-wise linear dependence of magnetization on field to characterize the magnetic beads. We implemented a bi-linear interpolation of magnetic field in the work space, based on a lookup table obtained from finite element simulation. The electronics and software were custom-made to achieve high performance. In addition, the effects of dimension and defect on structure of magnetic tips also were inspected. In a workspace with size of 0.1 × 0.1 mm{sup 2}, a force of up to 400 pN can be applied on a 2.8 μm superparamagnetic bead in any direction within the plane. Because the magnetic particle is always pulled towards a tip, the pulling forces from the pole tips have to be well balanced in order to achieve control of the particle’s position. Active video tracking based feedback control is implemented, which is able to work at a speed of up to 1 kHz, yielding good maneuverability of the magnetic beads.

Self-assembly of a superparamagnetic raspberry-like silica/iron oxide nanocomposite using epoxy-amine coupling chemistry.

Science.gov (United States)

Cano, Manuel; de la Cueva-Méndez, Guillermo

2015-02-28

The fabrication of colloidal nanocomposites would benefit from controlled hetero-assembly of ready-made particles through covalent bonding. Here we used epoxy-amine coupling chemistry to promote the self-assembly of superparamagnetic raspberry-like nanocomposites. This adaptable method induced the covalent attachment of iron oxide nanoparticles sparsely coated with amine groups onto epoxylated silica cores in the absence of other reactants.

Improving surface-enhanced Raman scattering effect using gold-coated hierarchical polystyrene bead substrates modified with postgrowth microwave treatment.

Science.gov (United States)

Yuen, Clement; Zheng, Wei; Huang, Zhiwei

2008-01-01

We report a novel postgrowth microwave heating implementation by selectively modifying hierarchical polystyrene (PS) bead substrates coated with gold (Au) films to effectively improve the surface-enhanced Raman scattering (SERS) effect on the analytes. The SERS signal of probe molecule rhodamine 6G (Rh 6G) on the microwave-treated Au-PS substrates can be improved by 10-fold, while the detection limit of Rh 6G in concentration can be enhanced by two orders of magnitude compared to the as-growth substrates. The high-quality SERS spectrum of saliva can also be acquired using the modified substrates, demonstrating the potential for the realization of the high-performance SERS substrates for biomedical applications.

Genotoxicity of Superparamagnetic Iron Oxide Nanoparticles in Granulosa Cells

Directory of Open Access Journals (Sweden)

Marina Pöttler

2015-11-01

Full Text Available Nanoparticles that are aimed at targeting cancer cells, but sparing healthy tissue provide an attractive platform of implementation for hyperthermia or as carriers of chemotherapeutics. According to the literature, diverse effects of nanoparticles relating to mammalian reproductive tissue are described. To address the impact of nanoparticles on cyto- and genotoxicity concerning the reproductive system, we examined the effect of superparamagnetic iron oxide nanoparticles (SPIONs on granulosa cells, which are very important for ovarian function and female fertility. Human granulosa cells (HLG-5 were treated with SPIONs, either coated with lauric acid (SEONLA only, or additionally with a protein corona of bovine serum albumin (BSA; SEONLA-BSA, or with dextran (SEONDEX. Both micronuclei testing and the detection of γH2A.X revealed no genotoxic effects of SEONLA-BSA, SEONDEX or SEONLA. Thus, it was demonstrated that different coatings of SPIONs improve biocompatibility, especially in terms of genotoxicity towards cells of the reproductive system.

Evaluation of umbilical cord mesenchymal stem cells labeling with superparamagnetic iron oxide nanoparticles coated with dextran and complexed with Poly-L-Lysine; Avaliacao da marcacao de celulas-tronco mesenquimais de cordao umbilical com nanoparticulas superparamagneticas de oxido de ferro recobertas com Dextran e complexadas a Poli-L-Lisina

Energy Technology Data Exchange (ETDEWEB)

Sibov, Tatiana Tais; Mamani, Javier Bustamante; Pavon, Lorena Favaro; Cardenas, Walter Humberto; Gamarra, Lionel Fernel, E-mail: tatianats@einstein.br [Instituto do Cerebro - InCe, Hospital Israelita Albert Einstein - HIAE, Sao Paulo, SP (Brazil); Miyaki, Liza Aya Mabuchi [Faculdade de Enfermagem, Hospital Israelita Albert Einstein - HIAE, Sao Paulo, SP (Brazil); Marti, Luciana Cavalheiro; Sardinha, Luiz Roberto [Centro de Pesquisa Experimental, Hospital Israelita Albert Einstein - HIAE, Sao Paulo, SP (Brazil); Oliveira, Daniela Mara de [Universidade de Brasilia - UnB, Brasilia, DF (Brazil)

2012-04-15

Objective: The objective of this study was to evaluate the effect of the labeling of umbilical cord vein derived mesenchymal stem cells with superparamagnetic iron oxide nanoparticles coated with dextran and complexed to a non-viral transfector agent transfector poly-L-lysine. Methods: The labeling of mesenchymal stem cells was performed using the superparamagnetic iron oxide nanoparticles/dextran complexed and not complexed to poly-L-lysine. Superparamagnetic iron oxide nanoparticles/dextran was incubated with poly-L-lysine in an ultrasonic sonicator at 37 deg C for 10 minutes for complex formation superparamagnetic iron oxide nanoparticles/dextran/poly-L-lysine by electrostatic interaction. Then, the mesenchymal stem cells were incubated overnight with the complex superparamagnetic iron oxide nanoparticles/dextran/poly-L-lysine and superparamagnetic iron oxide nanoparticles/dextran. After the incubation period the mesenchymal stem cells were evaluated by internalization of the complex superparamagnetic iron oxide nanoparticles/dextran/polyL-lysine and superparamagnetic iron oxide nanoparticles/dextran by Prussian Blue stain. Cellular viability of labeled mesenchymal stem cells was evaluated by cellular proliferation assay using 5,6-carboxyfluorescein-succinimidyl ester method and apoptosis detection by Annexin V- Propidium Iodide assay. Results: mesenchymal stem cells labeled with superparamagnetic iron oxide nanoparticles/ dextran without poly-L-lysine not internalized efficiently the superparamagnetic iron oxide nanoparticles due to its low presence detected within cells. Mesenchymal stem cells labeled with the complex superparamagnetic iron oxide nanoparticles/dextran/polyL-lysine efficiently internalized the superparamagnetic iron oxide nanoparticles due to greater presence in the cells interior. The viability and apoptosis assays demonstrated that the mesenchymal stem cells labeled and not labeled respectively with the superparamagnetic iron oxide

TiO2 beads and TiO2-chitosan beads for urease immobilization

International Nuclear Information System (INIS)

Ispirli Doğaç, Yasemin; Deveci, İlyas; Teke, Mustafa; Mercimek, Bedrettin

2014-01-01

The aim of the present study is to synthesize TiO 2 beads for urease immobilization. Two different strategies were used to immobilize the urease on TiO 2 beads. In the first method (A), urease enzyme was immobilized onto TiO 2 beads by adsorption and then crosslinking. In the second method (B), TiO 2 beads were coated with chitosan-urease mixture. To determine optimum conditions of immobilization, different parameters were investigated. The parameters of optimization were initial enzyme concentration (0.5; 1; 1.5; 2 mg/ml), alginate concentration (1; 2; 3%), glutaraldehyde concentration (1; 2; 3% v/v) and chitosan concentration (2; 3; 4 mg/ml). The optimum enzyme concentrations were determined as 1.5 mg/ml for A and 1.0 mg/ml for B. The other optimum conditions were found 2.0% (w/v) for alginate concentration (both A and B); 3.0 mg/ml for chitosan concentration (B) and 2.0% (v/v) for glutaraldehyde concentration (A). The optimum temperature (20-60 °C), optimum pH (3.0-10.0), kinetic parameters, thermal stability (4–70 °C), pH stability (4.0-9.0), operational stability (0-230 min) and reusability (20 times) were investigated for characterization. The optimum temperatures were 30 °C (A), 40 °C (B) and 35 °C (soluble). The temperature profiles of the immobilized ureases were spread over a large area. The optimum pH values for the soluble urease and immobilized urease prepared by using methods (A) and (B) were found to be 7.5, 7.0, 7.0, respectively. The thermal stabilities of immobilized enzyme sets were studied and they maintained 50% activity at 65 °C. However, at this temperature free urease protected only 15% activity. - Highlights: • TiO 2 and TiO 2 -chitosan beads for urease immobilization have been prepared and characterized. • The beads used in this work are good matrices for the immobilization of urease. • The immobilized urease was shown to have good properties and stabilities (pH and thermal stability, operational stability). • The 50

Magnetic Bead Based Immunoassay for Autonomous Detection of Toxins

Energy Technology Data Exchange (ETDEWEB)

Kwon, Y; Hara, C A; Knize, M G; Hwang, M H; Venkatesteswaran, K S; Wheeler, E K; Bell, P M; Renzi, R F; Fruetel, J A; Bailey, C G

2008-05-01

As a step towards toward the development of a rapid, reliable analyzer for bioagents in the environment, we are developing an automated system for the simultaneous detection of a group of select agents and toxins. To detect toxins, we modified and automated an antibody-based approach previously developed for manual medical diagnostics that uses fluorescent eTag{trademark} reporter molecules and is suitable for highly multiplexed assays. Detection is based on two antibodies binding simultaneously to a single antigen, one of which is labeled with biotin while the other is conjugated to a fluorescent eTag{trademark} through a cleavable linkage. Aqueous samples are incubated with the mixture of antibodies along with streptavidin-coated magnetic beads coupled to a photo-activatable porphyrin complex. In the presence of antigen, a molecular complex is formed where the cleavable linkage is held in proximity to the photoactivable group. Upon excitation at 680 nm, free radicals are generated, which diffuse and cleave the linkage, releasing the eTags{trademark}. Released eTags{trademark} are analyzed using capillary gel electrophoresis with laser-induced fluorescence detection. Limits of detection for ovalbumin and botulinum toxoid individually were 4 ng/mL (or 80 pg) and 16 ng/mL (or 320 pg), respectively, using the manual assay. In addition, we demonstrated the use of pairs of antibodies from different sources in a single assay to decrease the rate of false positives. Automation of the assay was demonstrated on a flow-through format with higher LODs of 125 ng/mL (or 2.5 ng) each of a mixture of ovalbumin and botulinum toxoid. This versatile assay can be easily modified with the appropriate antibodies to detect a wide range of toxins and other proteins.

Design and synthesis of biotin analogues reversibly binding with streptavidin.

Science.gov (United States)

Yamamoto, Tomohiro; Aoki, Kiyoshi; Sugiyama, Akira; Doi, Hirofumi; Kodama, Tatsuhiko; Shimizu, Yohei; Kanai, Motomu

2015-04-01

Two new biotin analogues, biotin carbonate 5 and biotin carbamate 6, have been synthesized. These molecules were designed to reversibly bind with streptavidin by replacing the hydrogen-bond donor NH group(s) of biotin's cyclic urea moiety with oxygen. Biotin carbonate 5 was synthesized from L-arabinose (7), which furnishes the desired stereochemistry at the 3,4-cis-dihydroxy groups, in 11% overall yield (over 10 steps). Synthesis of biotin carbamate 6 was accomplished from L-cysteine-derived chiral aldehyde 33 in 11% overall yield (over 7 steps). Surface plasmon resonance analysis of water-soluble biotin carbonate analogue 46 and biotin carbamate analogue 47 revealed that KD values of these compounds for binding to streptavidin were 6.7×10(-6)  M and 1.7×10(-10)  M, respectively. These values were remarkably greater than that of biotin (KD =10(-15)  M), and thus indicate the importance of the nitrogen atoms for the strong binding between biotin and streptavidin. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Monolayer-crystal streptavidin support films provide an internal standard of cryo-EM image quality

Energy Technology Data Exchange (ETDEWEB)

Han, Bong-Gyoon [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Watson, Zoe [Univ. of California, Berkeley, CA (United States); Cate, Jamie H. D. [Univ. of California, Berkeley, CA (United States); Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Glaeser, Robert M. [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)

2017-03-01

Analysis of images of biotinylated Escherichia coli 70S ribosome particles, bound to streptavidin affinity grids, demonstrates that the image-quality of particles can be predicted by the image-quality of the monolayer crystalline support film. Also, the quality of the Thon rings is a good predictor of the image-quality of particles, but only when images of the streptavidin crystals extend to relatively high resolution. When the estimated resolution of streptavidin was 5 Å or worse, for example, the ribosomal density map obtained from 22,697 particles went to only 9.5 Å, while the resolution of the map reached 4.0 Å for the same number of particles, when the estimated resolution of streptavidin crystal was 4 Å or better. It thus is easy to tell which images in a data set ought to be retained for further work, based on the highest resolution seen for Bragg peaks in the computed Fourier transforms of the streptavidin component. The refined density map obtained from 57,826 particles obtained in this way extended to 3.6 Å, a marked improvement over the value of 3.9 Å obtained previously from a subset of 52,433 particles obtained from the same initial data set of 101,213 particles after 3-D classification. These results are consistent with the hypothesis that interaction with the air-water interface can damage particles when the sample becomes too thin. Finally, streptavidin monolayer crystals appear to provide a good indication of when that is the case.

Monodisperse measurement of the biotin-streptavidin interaction strength in a well-defined pulling geometry.

Directory of Open Access Journals (Sweden)

Steffen M Sedlak

Full Text Available The widely used interaction of the homotetramer streptavidin with the small molecule biotin has been intensively studied by force spectroscopy and has become a model system for receptor ligand interaction. However, streptavidin's tetravalency results in diverse force propagation pathways through the different binding interfaces. This multiplicity gives rise to polydisperse force spectroscopy data. Here, we present an engineered monovalent streptavidin tetramer with a single cysteine in its functional subunit that allows for site-specific immobilization of the molecule, orthogonal to biotin binding. Functionality of streptavidin and its binding properties for biotin remain unaffected. We thus created a stable and reliable molecular anchor with a unique high-affinity binding site for biotinylated molecules or nanoparticles, which we expect to be useful for many single-molecule applications. To characterize the mechanical properties of the bond between biotin and our monovalent streptavidin, we performed force spectroscopy experiments using an atomic force microscope. We were able to conduct measurements at the single-molecule level with 1:1-stoichiometry and a well-defined geometry, in which force exclusively propagates through a single subunit of the streptavidin tetramer. For different force loading rates, we obtained narrow force distributions of the bond rupture forces ranging from 200 pN at 1,500 pN/s to 230 pN at 110,000 pN/s. The data are in very good agreement with the standard Bell-Evans model with a single potential barrier at Δx0 = 0.38 nm and a zero-force off-rate koff,0 in the 10-6 s-1 range.

Monodisperse measurement of the biotin-streptavidin interaction strength in a well-defined pulling geometry.

Science.gov (United States)

Sedlak, Steffen M; Bauer, Magnus S; Kluger, Carleen; Schendel, Leonard C; Milles, Lukas F; Pippig, Diana A; Gaub, Hermann E

2017-01-01

The widely used interaction of the homotetramer streptavidin with the small molecule biotin has been intensively studied by force spectroscopy and has become a model system for receptor ligand interaction. However, streptavidin's tetravalency results in diverse force propagation pathways through the different binding interfaces. This multiplicity gives rise to polydisperse force spectroscopy data. Here, we present an engineered monovalent streptavidin tetramer with a single cysteine in its functional subunit that allows for site-specific immobilization of the molecule, orthogonal to biotin binding. Functionality of streptavidin and its binding properties for biotin remain unaffected. We thus created a stable and reliable molecular anchor with a unique high-affinity binding site for biotinylated molecules or nanoparticles, which we expect to be useful for many single-molecule applications. To characterize the mechanical properties of the bond between biotin and our monovalent streptavidin, we performed force spectroscopy experiments using an atomic force microscope. We were able to conduct measurements at the single-molecule level with 1:1-stoichiometry and a well-defined geometry, in which force exclusively propagates through a single subunit of the streptavidin tetramer. For different force loading rates, we obtained narrow force distributions of the bond rupture forces ranging from 200 pN at 1,500 pN/s to 230 pN at 110,000 pN/s. The data are in very good agreement with the standard Bell-Evans model with a single potential barrier at Δx0 = 0.38 nm and a zero-force off-rate koff,0 in the 10-6 s-1 range.

Crystal structure of core streptavidin determined from multi-wavelength anomalous diffraction of synchrotron radiation

International Nuclear Information System (INIS)

Hendrickson, W.A.; Paehler, A.; Smith, J.L.; Satow, Y.; Merritt, E.A.; Phizackerley, R.P.

1989-01-01

A three-dimensional crystal structure of the biotin-binding core of streptavidin has been determined at 3.1-angstrom resolution. The structure was analyzed from diffraction data measured at three wavelengths from a single crystal of the selenobiotinyl complex with streptavidin. Streptavidin is a tetramer with subunits arrayed in D 2 symmetry. Each protomer is an 8-stranded β-barrel with simple up-down topology. Biotin molecules are bound at one end of each barrel. This study demonstrates the effectiveness of multi-wavelength anomalous diffraction (MAD) procedures for macromolecular crystallography and provides a basis for detailed study of biotin-avidin interactions

Synthesis and characterization of Fe3O4 nanoparticles coated with fucan polysaccharides

International Nuclear Information System (INIS)

Silva, V.A.J.; Andrade, P.L.; Silva, M.P.C.; Bustamante D, A.; De Los Santos Valladares, Luis; Albino Aguiar, J.

2013-01-01

In this work we report the preparation of fucan-coated magnetite (Fe 3 O 4 ) nanoparticles by the co-precipitation method. These nanoparticles were characterized by scanning electron microscopy, transmission electron microscopy, X-ray diffraction (XRD), Fourier transform infrared (FT-IR) spectroscopy, Mössbauer spectroscopy and magnetic measurements. The nanoparticles showed quasi-spherical morphology with mean sizes around 10 nm. XRD and FT-IR confirmed the functionalization of the Fe 3 O 4 nanoparticles with the fucan polysaccharide. Room temperature magnetization measurements and Mössbauer spectroscopy showed that the nanoparticles exhibited superparamagnetic behavior at 300 K and the magnetic properties of the Fe 3 O 4 are partly screened by the coating preventing aggregation. - Highlights: • Syntheses of fucan-coated Fe 3 O 4 nanoparticles were made by co-precipitation method. • The efficiency of polysaccharide coated was analyzed by XRD and FT-IR. • The magnetic nanoparticles mean size was 10–20 nm. • The fucan-coated magnetite nanoparticles showed superparamagnetic behavior

Functionality screen of streptavidin mutants by non-denaturing SDS-PAGE using biotin-4-fluorescein.

Science.gov (United States)

Humbert, Nicolas; Ward, Thomas R

2008-01-01

Site-directed mutagenesis or directed evolution of proteins often leads to the production of inactive mutants. For streptavidin and related proteins, mutations may lead to the loss of their biotin-binding properties. With high-throughput screening methodologies in mind, it is imperative to detect, prior to the high-density protein production, the bacteria that produce non-functional streptavidin isoforms. Based on the incorporation of biotin-4-fluorescein in streptavidin mutants present in Escherichia coli bacterial extracts, we detail a functional screen that allows the identification of biotin-binding streptavidin variants. Bacteria are cultivated in a small volume, followed by a rapid treatment of the cells; biotin-4-fluorescein is added to the bacterial extract and loaded on an Sodium Dodecyl Sulfate Poly-Acrylamide Gel Electrophoresis (SDS-PAGE) under non-denaturing conditions. Revealing is performed using a UV transilluminator. This screen is thus easy to implement, cheap and requires only readily available equipment.

Beads.

Science.gov (United States)

Weewish Tree, 1979

1979-01-01

Beads served both as ornaments and as a medium of exchange, and the Indians manufactured them from such natural sources as bones, stones, beans, nuts, animal teeth, and polished antlers. Even after the introduction of European glass beads, the Indians continued to make their favorite beads from the natural sources. (DS)

A streptavidin linker layer that functions after drying.

Science.gov (United States)

Xia, Nan; Shumaker-Parry, Jennifer S; Zareie, M Hadi; Campbell, Charles T; Castner, David G

2004-04-27

The ability of streptavidin (SA) to simultaneously bind four biotins is often used in linker layers, where a biotinylated molecule is linked to a biotin-functionalized surface via SA. For biosensor and array applications, it is desirable that the SA linker layer be stable to drying and rehydration. In this study it was observed that a significant decrease in binding capacity of a SA layer occurred when that layer was dried. For this study a SA linker layer was constructed by binding SA to a biotin-containing alkylthiolate monolayer (BAT/OEG) self-assembled onto gold. Its stability after drying was investigated using surface plasmon resonance (SPR). Approximately a quarter of the SA layer was removed from the BAT/OEG surface upon drying and rehydration, suggesting disruption of SA-biotin binding when dry. This resulted in the dried SA layer losing approximately 40% of its biotinylated ferritin (BF) binding capacity. Coating the layer with trehalose before drying was found to inhibit the loss of SA from the BAT/OEG surface. SPR showed that the trehalose-protected SA linker layer retained approximately 91% of its original BF binding capacity after drying and rehydration. Atomic force microscopy, which was used to image individual surface-bound SA and BF molecules, qualitatively confirmed these observations.

The growth speed of microtubules with XMAP215-coated beads coupled to their ends is increased by tensile force

Science.gov (United States)

Trushko, Anastasiya; Schäffer, Erik; Howard, Jonathon

2013-01-01

The generation of pulling and pushing forces is one of the important functions of microtubules, which are dynamic and polarized structures. The ends of dynamic microtubules are able to form relatively stable links to cellular structures, so that when a microtubule grows it can exert a pushing force and when it shrinks it can exert a pulling force. Microtubule growth and shrinkage are tightly regulated by microtubule-associated proteins (MAPs) that bind to microtubule ends. Given their localization, MAPs may be exposed to compressive and tensile forces. The effect of such forces on MAP function, however, is poorly understood. Here we show that beads coated with the microtubule polymerizing protein XMAP215, the Xenopus homolog of Dis1 and chTOG, are able to link stably to the plus ends of microtubules, even when the ends are growing or shrinking; at growing ends, the beads increase the polymerization rate. Using optical tweezers, we found that tensile force further increased the microtubule polymerization rate. These results show that physical forces can regulate the activity of MAPs. Furthermore, our results show that XMAP215 can be used as a handle to sense and mechanically manipulate the dynamics of the microtubule tip. PMID:23964126

Environmentally Compatible Synthesis of Superparamagnetic Magnetite (Fe3O4 Nanoparticles with Prehydrolysate from Corn Stover

Directory of Open Access Journals (Sweden)

Chunming Zheng

2013-12-01

Full Text Available An environmentally compatible and size-controlled method has been employed for synthesis of superparamagnetic magnetite nanoparticles with prehydrolysate from corn stover. Various characterizations involving X-ray diffraction (XRD, standard and high-resolution transmission electron microscopy (TEM and HRTEM, selected area electron diffraction (SAED, and thermogravimetric analysis (TGA have integrally confirmed the formation of magnetite nanoparticles with homogeneous morphology and the formation mechanism of magnetite only from ferric precursor. Organic materials in the prehydrolysate act as a bifunctional agent: (1 a reducing agent to reduce ferric ions to prepare magnetite with the coexistence of ferric and ferrous ions; and (2 a coating agent to prevent particle growth and agglomeration and to promote the formation of nanoscale and superparamagnetic magnetite. The size of the magnetite nanoparticles can be easily controlled by tailoring the reducing sugar concentration, reaction time, or hydrothermal temperature.

Magnetic separation of encapsulated islet cells labeled with superparamagnetic iron oxide nano particles.

Science.gov (United States)

Mettler, Esther; Trenkler, Anja; Feilen, Peter J; Wiegand, Frederik; Fottner, Christian; Ehrhart, Friederike; Zimmermann, Heiko; Hwang, Yong Hwa; Lee, Dong Yun; Fischer, Stefan; Schreiber, Laura M; Weber, Matthias M

2013-01-01

Islet cell transplantation is a promising option for the restoration of normal glucose homeostasis in patients with type 1 diabetes. Because graft volume is a crucial issue in islet transplantations for patients with diabetes, we evaluated a new method for increasing functional tissue yield in xenogeneic grafts of encapsulated islets. Islets were labeled with three different superparamagnetic iron oxide nano particles (SPIONs; dextran-coated SPION, siloxane-coated SPION, and heparin-coated SPION). Magnetic separation was performed to separate encapsulated islets from the empty capsules, and cell viability and function were tested. Islets labeled with 1000 μg Fe/ml dextran-coated SPIONs experienced a 69.9% reduction in graft volume, with a 33.2% loss of islet-containing capsules. Islets labeled with 100 μg Fe/ml heparin-coated SPIONs showed a 46.4% reduction in graft volume, with a 4.5% loss of capsules containing islets. No purification could be achieved using siloxane-coated SPIONs due to its toxicity to the primary islets. SPION labeling of islets is useful for transplant purification during islet separation as well as in vivo imaging after transplantation. Furthermore, purification of encapsulated islets can also reduce the volume of the encapsulated islets without impairing their function by removing empty capsules. © 2013 John Wiley & Sons A/S.

Multiplex detection of plant pathogens through the luminex magplex bead system

NARCIS (Netherlands)

Vlugt, van der R.A.A.; Raaij, van H.M.G.; Weerdt, de M.; Bergervoet, J.H.W.

2015-01-01

Here we describe a versatile multiplex method for both the serological and molecular detection of plant pathogens. The Luminex MagPlex bead system uses small paramagnetic microspheres (“beads”), either coated with specific antibodies or oligonucleotides, which capture respectively viruses and/or

Uptake of magnetic nanoparticles into cells for cell tracking

International Nuclear Information System (INIS)

Becker, Christiane; Hodenius, Michael; Blendinger, Gitta; Sechi, Antonio; Hieronymus, Thomas; Mueller-Schulte, Detlef; Schmitz-Rode, Thomas; Zenke, Martin

2007-01-01

A challenge for future applications in nanotechnology is the functional integration of nano-sized materials into cellular structures. Here we investigated superparamagnetic Fe 3 O 4 iron oxide nanoparticles coated with a lipid bilayer for uptake into cells and for targeting subcellular compartments. It was found that magnetic nanoparticles (MNPs) are effectively taken up into cells and make cells acquire magnetic activity. Biotin-conjugated MNPs were further functionalized by binding of the fluorescent tag streptavidin-fluorescein isothiocyanate (FITC) and, following uptake into cells, shown to confer magnetic activity and fluorescence labeling. Such FITC-MNPs were localized in the lysosomal compartment of cells which suggests a receptor-mediated uptake mechanism

Engineering of a Bacillus subtilis strain with adjustable levels of intracellular biotin for secretory production of functional streptavidin.

Science.gov (United States)

Wu, Sau-Ching; Wong, Sui-Lam

2002-03-01

Streptavidin is a biotin-binding protein which has been widely used in many in vitro and in vivo applications. Because of the ease of protein recovery and availability of protease-deficient strains, the Bacillus subtilis expression-secretion system is an attractive system for streptavidin production. However, attempts to produce streptavidin using B. subtilis face the problem that cells overproducing large amounts of streptavidin suffer poor growth, presumably because of biotin deficiency. This problem cannot be solved by supplementing biotin to the culture medium, as this will saturate the biotin binding sites in streptavidin. We addressed this dilemma by engineering a B. subtilis strain (WB800BIO) which overproduces intracellular biotin. The strategy involves replacing the natural regulatory region of the B. subtilis chromosomal biotin biosynthetic operon (bioWAFDBIorf2) with an engineered one consisting of the B. subtilis groE promoter and gluconate operator. Biotin production in WB800BIO is induced by gluconate, and the level of biotin produced can be adjusted by varying the gluconate dosage. A level of gluconate was selected to allow enhanced intracellular production of biotin without getting it released into the culture medium. WB800BIO, when used as a host for streptavidin production, grows healthily in a biotin-limited medium and produces large amounts (35 to 50 mg/liter) of streptavidin, with over 80% of its biotin binding sites available for future applications.

Binding of human endothelium to Ulex europaeus I-coated Dynabeads: application to the isolation of microvascular endothelium.

Science.gov (United States)

Jackson, C J; Garbett, P K; Nissen, B; Schrieber, L

1990-06-01

A major problem encountered when isolating human microvascular endothelium is the presence of contaminating cells such as fibroblasts that rapidly over-grow the endothelial cells. We describe here a simple, rapid technique for purifying endothelial cells derived from the microvasculature of neonatal foreskin and osteoarthritic and rheumatoid arthritic synovium. This technique is based on the selective binding of the lectin Ulex europaeus I (UEA I) to the endothelial cell surface via fucose residues. Initially UEA I was covalently bound to tosyl-activated super-paramagnetic polystyrene beads (Dynabeads) by incubation for 24 h at room temperature. Cells were isolated by extracting microvascular segments from enzyme-treated (trypsin and Pronase) cubes of tissue. The mixed population of cells obtained were purified by incubating them at 4 degrees C for 10 min with the UEA I-coated Dynabeads. Endothelium bound to the beads whilst contaminating cells were removed by five washes with HBSS using a magnetic particle concentrator. The endothelial cells thus obtained grew to confluence as a cobblestone-like monolayer and expressed von Willebrand factor antigen. The cells were released from the Dynabeads by the competitive binding of fucose (10 min at 4 degrees C). This new method is simple and reproducible and allows pure human microvascular endothelial cells to be cultured within 2 h of obtaining a specimen.

Efficient synthesis of superparamagnetic magnetite nanoparticles under air for biomedical applications

Energy Technology Data Exchange (ETDEWEB)

Saxena, Namita, E-mail: saxenanamita@yahoo.com [School of Nano Sciences, Central University of Gujarat, Gandhinagar 382030 (India); Singh, Man, E-mail: mansingh50@hotmail.com [School of Chemical Sciences, Central University of Gujarat, Gandhinagar 382030 (India)

2017-05-01

The facile co-precipitation process of synthesising Superparamagnetic Iron Oxide Nanoparticles (SPIONs) especially magnetite was investigated and simplified, to develop a reproducible and scaled up synthesis process under air, for producing particles with enhanced percentage of magnetite, thus eliminating the crucial and complicated need of using the inert atmosphere. Presence of magnetite was confirmed by XRD, TEM, and Raman spectroscopy. Efficiency of synthesising magnetite was increased up to approx. ∼58 wt%, under air with no other phases but maghemite present. Alkali concentration was optimised, and particles with better magnetisation values were synthesised. The approximate weight percentage of magnetite was calculated using the simple and rapid XRD peak deconvolution method. Higher pH values from 13 to14 were investigated in the study while alkali concentration was varied from 0.5 to 4 M. 1Molar NaOH with a final pH of 13.4 was found to be optimum. Well crystallised particles with approx. 6–12 nm size, narrow size distribution and cubo-spheroidal shape were synthesised. Particles were Superparamagnetic with high values of saturation magnetisation of up to 68 emu/g and negligible values of remanence and coercivity. A reaction yield of up to 62% was obtained. Hydrophilic coated particles were produced in a single, one step facile process for biomedical applications, using optimised parameters of pH and alkali concentration obtained in the study. Single domain particles with good magnetisation formed stable aqueous dispersions. FTIR, UV-Visible and DLS were used to confirm the coating and dispersion stabilities of the particles. These particles have the requisite properties required for application in different biomedical fields.

Intraarticular application of superparamagnetic nanoparticles and their uptake by synovial membrane-an experimental study in sheep

Energy Technology Data Exchange (ETDEWEB)

Schulze, Katja [Musculoskeletal Research Unit, Equine Hospital, Vetsuisse Faculty, University of Zurich, Winterthurerstrasse 260, 8057 Zurich (Switzerland); Koch, Annette [Department of Chemistry and Applied BioSciences, Swiss Federal Institute of Technology Zurich (ETH Zurich), Winterthurerstrasse 190, 8057 Zurich (Switzerland); Schoepf, Bernhard [Musculoskeletal Research Unit, Equine Hospital, Vetsuisse Faculty, University of Zurich, Winterthurerstrasse 260, 8057 Zurich (Switzerland); Petri, Alke [Laboratory of Powder Technology, Swiss Federal Institute of Technology (EPFL), Lausanne (Switzerland); Steitz, Benedikt [Laboratory of Powder Technology, Swiss Federal Institute of Technology (EPFL), Lausanne (Switzerland); Chastellain, Mathieu [Laboratory of Powder Technology, Swiss Federal Institute of Technology (EPFL), Lausanne (Switzerland); Hofmann, Margarethe [MatSearch, Chemin Jean Pavillard 14, 1009 Pully (Switzerland); Hofmann, Heinrich [Laboratory of Powder Technology, Swiss Federal Institute of Technology (EPFL), Lausanne (Switzerland); Rechenberg, Brigitte von [Musculoskeletal Research Unit, Equine Hospital, Vetsuisse Faculty, University of Zurich, Winterthurerstrasse 260, 8057 Zurich (Switzerland)]. E-mail: bvonrechenberg@vetclinics.unizh.ch

2005-05-15

A superparamagnetic iron oxide nanoparticle, coated with polyvinyl alcohol (PVA-SPION) and its fluorescently functionalized analogue (amino-PVA-Cy3.5-SPION) were compared in vivo as proof of principle for future use in magnetic drug targeting in inflammatory joint diseases. They were injected either intraarticularly or periarticularly and their uptake by cells of the synovial membrane was evaluated. Uptake was completed in 48 h and was enforced by an extracorporally applied magnet.

Intraarticular application of superparamagnetic nanoparticles and their uptake by synovial membrane-an experimental study in sheep

International Nuclear Information System (INIS)

Schulze, Katja; Koch, Annette; Schoepf, Bernhard; Petri, Alke; Steitz, Benedikt; Chastellain, Mathieu; Hofmann, Margarethe; Hofmann, Heinrich; Rechenberg, Brigitte von

2005-01-01

A superparamagnetic iron oxide nanoparticle, coated with polyvinyl alcohol (PVA-SPION) and its fluorescently functionalized analogue (amino-PVA-Cy3.5-SPION) were compared in vivo as proof of principle for future use in magnetic drug targeting in inflammatory joint diseases. They were injected either intraarticularly or periarticularly and their uptake by cells of the synovial membrane was evaluated. Uptake was completed in 48 h and was enforced by an extracorporally applied magnet

Comparison of anti-CD3 and anti-CD28-coated beads with soluble anti-CD3 for expanding human T cells: Differing impact on CD8 T cell phenotype and responsiveness to restimulation

Directory of Open Access Journals (Sweden)

Kurlander Roger J

2010-10-01

Full Text Available Abstract Background The ability to expand virus- or tumor-specific T cells without damaging their functional capabilities is critical for success adoptive transfer immunotherapy of patients with opportunistic infection or tumor. Careful comparisons can help identify expansion methods better suited for particular clinical settings and identify recurrent deficiencies requiring new innovation. Methods We compared the efficacy of magnetic beads coated with anti-CD3 and anti-CD28 (anti-CD3/CD28 beads, and soluble anti-CD3 plus mixed mononuclear cells (designated a rapid expansion protocol or REP in expanding normal human T cells. Results Both anti-CD3/CD28 beads and soluble anti-CD3 promoted extensive expansion. Beads stimulated greater CD4 cell growth (geometric mean of 56- versus 27-fold (p Conclusions Anti-CD3/CD28 beads are highly effective for expanding CD4 cells, but soluble anti-CD3 has significant potential advantages for expanding CD8 T cells, particularly where preservation of phenotypically "young" CD8 cells would be desirable, or where the T cells of interest have been antigen-stimulated in vitro or in vivo in the recent past.

Design of Superparamagnetic Nanoparticles for Magnetic Particle Imaging (MPI

Directory of Open Access Journals (Sweden)

Philip W. T. Pong

2013-09-01

Full Text Available Magnetic particle imaging (MPI is a promising medical imaging technique producing quantitative images of the distribution of tracer materials (superparamagnetic nanoparticles without interference from the anatomical background of the imaging objects (either phantoms or lab animals. Theoretically, the MPI platform can image with relatively high temporal and spatial resolution and sensitivity. In practice, the quality of the MPI images hinges on both the applied magnetic field and the properties of the tracer nanoparticles. Langevin theory can model the performance of superparamagnetic nanoparticles and predict the crucial influence of nanoparticle core size on the MPI signal. In addition, the core size distribution, anisotropy of the magnetic core and surface modification of the superparamagnetic nanoparticles also determine the spatial resolution and sensitivity of the MPI images. As a result, through rational design of superparamagnetic nanoparticles, the performance of MPI could be effectively optimized. In this review, the performance of superparamagnetic nanoparticles in MPI is investigated. Rational synthesis and modification of superparamagnetic nanoparticles are discussed and summarized. The potential medical application areas for MPI, including cardiovascular system, oncology, stem cell tracking and immune related imaging are also analyzed and forecasted.

Time-resolved homo-FRET studies of biotin-streptavidin complexes.

Science.gov (United States)

Andreoni, Alessandra; Nardo, Luca; Rigler, Rudolf

2016-09-01

Förster resonance energy transfer is a mechanism of fluorescence quenching that is notably useful for characterizing properties of biomolecules and/or their interactions. Here we study water-solutions of Biotin-Streptavidin complexes, in which Biotin is labeled with a rigidly-bound fluorophore that can interact by Förster resonance energy transfer with the fluorophores labeling the other, up to three, Biotins of the same complex. The fluorophore, Atto550, is a Rhodamine analogue. We detect the time-resolved fluorescence decay of the fluorophores with an apparatus endowed with single-photon sensitivity and temporal resolution of ~30ps. The decay profiles we observe for samples containing constant Biotin-Atto550 conjugates and varying Streptavidin concentrations are multi-exponential. Each decay component can be associated with the rate of quenching exerted on each donor by each of the acceptors that label the other Biotin molecules, depending on the binding site they occupy. The main features that lead to this result are that (i) the transition dipole moments of the up-to-four Atto550 fluorophores that label the complexes are fixed as to both relative positions and mutual orientations; (ii) the fluorophores are identical and the role of donor in each Biotin-Streptavidin complex is randomly attributed to the one that has absorbed the excitation light (homo-FRET). Obviously the high-temporal resolution of the excitation-detection apparatus is necessary to discriminate among the fluorescence decay components. Copyright © 2016 Elsevier B.V. All rights reserved.

Synthesis of Superparamagnetic Iron Oxide Nanoparticles Modified with MPEG-PEI via Photochemistry as New MRI Contrast Agent

Directory of Open Access Journals (Sweden)

Yancong Zhang

2015-01-01

Full Text Available Novel method for synthesis of superparamagnetic iron oxide nanoparticles (SPIONs coated with polyethylenimine (PEI and modified with poly(ethylene glycol methyl ether (MPEG, MPEG-PEI-SPIONs, was developed. PEI-SPIONs were successfully prepared in aqueous system via photochemistry, and their surface was modified with poly(ethylene glycol methyl ether (MPEG. The so-obtained MPEG-PEI-SPIONs had a uniform hydrodynamic particle size of 34 nm. The successful coating of MPEG-PEI on the SPIONs was ascertained from FT-IR analysis, and the PEI and MPEG fractions in MPEG-PEI-SPIONs were calculated to account for 31% and 12%, respectively. Magnetic measurement revealed that the saturated magnetization of MPEG-PEI-SPIONs reached 46 emu/g and the nanoparticles showed the characteristic of being superparamagnetic. The stability experiment revealed that the MPEG-PEI modification improved the nanoparticles stability greatly. T2 relaxation measurements showed that MPEG-PEI-SPIONs show similar R2 value to the PEI-SPIONs. The T2-weighted magnetic resonance imaging (MRI of MPEG-PEI-SPIONs showed that the magnetic resonance signal was enhanced significantly with increasing nanoparticle concentration in water. These results indicated that the MPEG-PEI-SPIONs had great potential for application in MRI.

Connecting DNA Origami Structures Using the Biotin-Streptavidin ...

African Journals Online (AJOL)

Abstract. This work made use of the strong interaction between biotin and streptavidin to connect designed DNA origami structures. The caDNAno software was used to design a 6 layer 3D origami cross-like structure. Selected DNA strands at the engineered attachment sites on the DNA origami structure were biotinylated.

A disposable electrochemical immunosensor for prolactin involving affinity reaction on streptavidin-functionalized magnetic particles

International Nuclear Information System (INIS)

Moreno-Guzman, Maria; Gonzalez-Cortes, Araceli; Yanez-Sedeno, Paloma; Pingarron, Jose M.

2011-01-01

A novel electrochemical immunosensor was developed for the determination of the hormone prolactin. The design involved the use of screen-printed carbon electrodes and streptavidin-functionalized magnetic particles. Biotinylated anti-prolactin antibodies were immobilized onto the functionalized magnetic particles and a sandwich-type immunoassay involving prolactin and anti-prolactin antibody labelled with alkaline phosphatase was employed. The resulting bio-conjugate was trapped on the surface of the screen-printed electrode with a small magnet and prolactin quantification was accomplished by differential pulse voltammetry of 1-naphtol formed in the enzyme reaction using 1-naphtyl phosphate as alkaline phosphatase substrate. All variables involved in the preparation of the immunosensor and in the electrochemical detection step were optimized. The calibration plot for prolactin exhibited a linear range between 10 and 2000 ng mL -1 with a slope value of 7.0 nA mL ng -1 . The limit of detection was 3.74 ng mL -1 . Furthermore, the modified magnetic beads-antiprolactin conjugates showed an excellent stability. The immunosensor exhibited also a high selectivity with respect to other hormones. The analytical usefulness of the immnunosensor was demonstrated by analyzing human sera spiked with prolactin at three different concentration levels.

A disposable electrochemical immunosensor for prolactin involving affinity reaction on streptavidin-functionalized magnetic particles

Energy Technology Data Exchange (ETDEWEB)

Moreno-Guzman, Maria; Gonzalez-Cortes, Araceli [Department of Analytical Chemistry, Faculty of Chemistry, University Computense of Madrid, 28040 Madrid (Spain); Yanez-Sedeno, Paloma, E-mail: yseo@quim.ucm.es [Department of Analytical Chemistry, Faculty of Chemistry, University Computense of Madrid, 28040 Madrid (Spain); Pingarron, Jose M. [Department of Analytical Chemistry, Faculty of Chemistry, University Computense of Madrid, 28040 Madrid (Spain)

2011-04-29

A novel electrochemical immunosensor was developed for the determination of the hormone prolactin. The design involved the use of screen-printed carbon electrodes and streptavidin-functionalized magnetic particles. Biotinylated anti-prolactin antibodies were immobilized onto the functionalized magnetic particles and a sandwich-type immunoassay involving prolactin and anti-prolactin antibody labelled with alkaline phosphatase was employed. The resulting bio-conjugate was trapped on the surface of the screen-printed electrode with a small magnet and prolactin quantification was accomplished by differential pulse voltammetry of 1-naphtol formed in the enzyme reaction using 1-naphtyl phosphate as alkaline phosphatase substrate. All variables involved in the preparation of the immunosensor and in the electrochemical detection step were optimized. The calibration plot for prolactin exhibited a linear range between 10 and 2000 ng mL{sup -1} with a slope value of 7.0 nA mL ng{sup -1}. The limit of detection was 3.74 ng mL{sup -1}. Furthermore, the modified magnetic beads-antiprolactin conjugates showed an excellent stability. The immunosensor exhibited also a high selectivity with respect to other hormones. The analytical usefulness of the immnunosensor was demonstrated by analyzing human sera spiked with prolactin at three different concentration levels.

High frequency lateral flow affinity assay using superparamagnetic nanoparticles

Energy Technology Data Exchange (ETDEWEB)

Lago-Cachón, D., E-mail: dlagocachon@gmail.com [Dpto. de Física, Universidad de Oviedo, Edificio Departamental Este, Campus de Viesques, 33204 Gijón (Spain); Rivas, M., E-mail: rivas@uniovi.es [Dpto. de Física, Universidad de Oviedo, Edificio Departamental Este, Campus de Viesques, 33204 Gijón (Spain); Martínez-García, J.C., E-mail: jcmg@uniovi.es [Dpto. de Física, Universidad de Oviedo, Edificio Departamental Este, Campus de Viesques, 33204 Gijón (Spain); Oliveira-Rodríguez, M., E-mail: oliveiramyriam@uniovi.es [Dpto. de Química Física y Analítica, Universidad de Oviedo, C/Julián Clavería 8, 33006 Oviedo (Spain); Blanco-López, M.C., E-mail: cblanco@uniovi.es [Dpto. de Química Física y Analítica, Universidad de Oviedo, C/Julián Clavería 8, 33006 Oviedo (Spain); García, J.A., E-mail: joseagd@uniovi.es [Dpto. de Física, Universidad de Oviedo, Escuela de Marina, Campus de Viesques, 33204 Gijón (Spain)

2017-02-01

Lateral flow assay is one of the simplest and most extended techniques in medical diagnosis for point-of-care testing. Although it has been traditionally a positive/negative test, some work has been lately done to add quantitative abilities to lateral flow assay. One of the most successful strategies involves magnetic beads and magnetic sensors. Recently, a new technique of superparamagnetic nanoparticle detection has been reported, based on the increase of the impedance induced by the nanoparticles on a RF-current carrying copper conductor. This method requires no external magnetic field, which reduces the system complexity. In this work, nitrocellulose membranes have been installed on the sensor, and impedance measurements have been carried out during the sample diffusion by capillarity along the membrane. The impedance of the sensor changes because of the presence of magnetic nanoparticles. The results prove the potentiality of the method for point-of-care testing of biochemical substances and nanoparticle capillarity flow studies. - Highlights: • A method for quantification of Lateral Flow Assays is proposed. • MNP induce an increase of the impedance on a RF-current carrying copper sensor. • Magnetic nanoparticles (MNP) can be detected flowing over the sensing element.

High frequency lateral flow affinity assay using superparamagnetic nanoparticles

International Nuclear Information System (INIS)

Lago-Cachón, D.; Rivas, M.; Martínez-García, J.C.; Oliveira-Rodríguez, M.; Blanco-López, M.C.; García, J.A.

2017-01-01

Lateral flow assay is one of the simplest and most extended techniques in medical diagnosis for point-of-care testing. Although it has been traditionally a positive/negative test, some work has been lately done to add quantitative abilities to lateral flow assay. One of the most successful strategies involves magnetic beads and magnetic sensors. Recently, a new technique of superparamagnetic nanoparticle detection has been reported, based on the increase of the impedance induced by the nanoparticles on a RF-current carrying copper conductor. This method requires no external magnetic field, which reduces the system complexity. In this work, nitrocellulose membranes have been installed on the sensor, and impedance measurements have been carried out during the sample diffusion by capillarity along the membrane. The impedance of the sensor changes because of the presence of magnetic nanoparticles. The results prove the potentiality of the method for point-of-care testing of biochemical substances and nanoparticle capillarity flow studies. - Highlights: • A method for quantification of Lateral Flow Assays is proposed. • MNP induce an increase of the impedance on a RF-current carrying copper sensor. • Magnetic nanoparticles (MNP) can be detected flowing over the sensing element.

A label-free colorimetric aptasensor for simple, sensitive and selective detection of Pt (II) based on platinum (II)-oligonucleotide coordination induced gold nanoparticles aggregation.

Science.gov (United States)

Fan, Daoqing; Zhai, Qingfeng; Zhou, Weijun; Zhu, Xiaoqing; Wang, Erkang; Dong, Shaojun

2016-11-15

Herein, a gold nanoparticles (AuNPs) based label-free colorimetric aptasensor for simple, sensitive and selective detection of Pt (II) was constructed for the first time. Four bases (G-G mismatch) mismatched streptavidin aptamer (MSAA) was used to protect AuNPs from salt-induced aggregation and recognize Pt (II) specifically. Only in the presence of Pt (II), coordination occurs between G-G bases and Pt (II), leading to the activation of streptavidin aptamer. Streptavidin coated magnetic beads (MBs) were used as separation agent to separate Pt (II)-coordinated MSAA. The residual less amount of MSAA could not efficiently protect AuNPs anymore and aggregation of AuNPs will produce a colorimetric product. With the addition of Pt (II), a pale purple-to-blue color variation could be observed by the naked eye. A detection limit of 150nM and a linear range from 0.6μM to 12.5μM for Pt (II) could be achieved without any amplification. Copyright © 2016 Elsevier B.V. All rights reserved.

Superparamagnetic photocurable nanocomposite for the fabrication of microcantilevers

DEFF Research Database (Denmark)

Suter, M; Ergeneman, O; Zürcher, J

2011-01-01

We present a photocurable polymer composite with superparamagnetic characteristics for the fabrication of microcantilevers. Uniform distribution and low particle agglomeration (......We present a photocurable polymer composite with superparamagnetic characteristics for the fabrication of microcantilevers. Uniform distribution and low particle agglomeration (...

Non-immunogenic dextran-coated superparamagnetic iron oxide nanoparticles: a biocompatible, size-tunable contrast agent for magnetic resonance imaging.

Science.gov (United States)

Unterweger, Harald; Janko, Christina; Schwarz, Marc; Dézsi, László; Urbanics, Rudolf; Matuszak, Jasmin; Őrfi, Erik; Fülöp, Tamás; Bäuerle, Tobias; Szebeni, János; Journé, Clément; Boccaccini, Aldo R; Alexiou, Christoph; Lyer, Stefan; Cicha, Iwona

2017-01-01

Iron oxide-based contrast agents have been in clinical use for magnetic resonance imaging (MRI) of lymph nodes, liver, intestines, and the cardiovascular system. Superparamagnetic iron oxide nanoparticles (SPIONs) have high potential as a contrast agent for MRI, but no intravenous iron oxide-containing agents are currently approved for clinical imaging. The aim of our work was to analyze the hemocompatibility and immuno-safety of a new type of dextran-coated SPIONs (SPIONdex) and to characterize these nanoparticles with ultra-high-field MRI. Key parameters related to nanoparticle hemocompatibility and immuno-safety were investigated in vitro and ex vivo. To address concerns associated with hypersensitivity reactions to injectable nanoparticulate agents, we analyzed complement activation-related pseudoallergy (CARPA) upon intravenous administration of SPIONdex in a pig model. Furthermore, the size-tunability of SPIONdex and the effects of size reduction on their biocompatibility were investigated. In vitro, SPIONdex did not induce hemolysis, complement or platelet activation, plasma coagulation, or leukocyte procoagulant activity, and had no relevant effect on endothelial cell viability or endothelial-monocytic cell interactions. Furthermore, SPIONdex did not induce CARPA even upon intravenous administration of 5 mg Fe/kg in pigs. Upon SPIONdex administration in mice, decreased liver signal intensity was observed after 15 minutes and was still detectable 24 h later. In addition, by changing synthesis parameters, a reduction in particle size contrast agent.

Repetitive Immunosensor with a Fiber-Optic Device and Antibody-Coated Magnetic Beads for Semi-Continuous Monitoring of Escherichia coli O157:H7.

Science.gov (United States)

Taniguchi, Midori; Saito, Hirokazu; Mitsubayashi, Kohji

2017-09-19

A rapid and reproducible fiber-optic immunosensor for Escherichia coli O157:H7 ( E. coli O157:H7) was described. The biosensor consisted of a flow cell, an optical fiber with a thin Ni layer, and a PC linked fluorometer. First, the samples with E. coli O157:H7 were incubated with magnetic beads coated with anti- E. coli O157:H7 antibodies and anti- E. coli O157:H7 antibodies labeled cyanine 5 (Cy5) to make sandwich complexes. Then the Cy5-( E. coli O157:H7)-beads were injected into a flow cell and pulled to the magnetized Ni layer on the optical fiber set in the flow cell. An excitation light (λ = 635 nm) was used to illuminate the optical fiber, and the Cy5 florescent molecules facing the optical fiber were exposed to an evanescent wave from the optical fiber. The 670 nm fluorescent light was measured using a photodiode. Finally, the magnetic intensity of the Ni layer was removed and the Cy5- E. coli O157:H7-beads were washed out for the next immunoassay. E. coli O157:H7, diluted with phosphate buffer (PB), was measured from 1 × 10⁵ to 1 × 10⁷ cells/mL. The total time required for an assay was less than 15 min (except for the pretreatment process) and repeating immunoassay on one optical fiber was made possible.

Certain patterns of IgG adsorption by polystyrene bead surfaces

Energy Technology Data Exchange (ETDEWEB)

Mamedov, M K

1985-01-01

The article reports on tests of domestic Soviet polystyrene beads that permit a simplified modification of the enzyme-adsorption method to identify the alpha hepatitis virus and its antibody in nonspecialized, general laboratories. Only patterns of Ig immunoglobulin adsorption were studied. Human IgG was conjugated with the radioactive isotope /sup 125/I by a chloramine method, with mean radioactivity and protein concentration measured frequently. Bovine serum albumin (BSA) and an anionic detergent Tween-20, and a phosphate-salt buffer with pH 5.8-8.2, were used to produce m-Ig and Ig. Adsorption involved incubation of the beads in various solutions, followed by measurement of their radioactivity. Results of several series of tests were subjected to Student-Fisher evaluation. This suggested that the presence of albumin in physiological concentrations in the solution had no important impact on m-Ig adsorption on the bead surface, which effectively adsorbed Ig from solutions without additional proteins, but also from Ig solutions containing serum albumin in physiological concentrations. Thus, it was possible to coat the beads with alpha Ig hepatitis virus. The Tween-20 weak detergent was effective for eliminating unwanted protein adsorption. 9 references, 3 figures.

Selection and Characterization of Single Stranded DNA Aptamers for the Hormone Abscisic Acid

Science.gov (United States)

Gonzalez, Victor M.; Millo, Enrico; Sturla, Laura; Vigliarolo, Tiziana; Bagnasco, Luca; Guida, Lucrezia; D'Arrigo, Cristina; De Flora, Antonio; Salis, Annalisa; Martin, Elena M.; Bellotti, Marta; Zocchi, Elena

2013-01-01

The hormone abscisic acid (ABA) is a small molecule involved in pivotal physiological functions in higher plants. Recently, ABA has been also identified as an endogenous hormone in mammals, regulating different cell functions including inflammatory processes, stem cell expansion, insulin release, and glucose uptake. Aptamers are short, single-stranded (ss) oligonucleotidesable to recognize target molecules with high affinity. The small size of the ABA molecule represented a challenge for aptamer development and the aim of this study was to develop specific anti-ABA DNA aptamers. Biotinylated abscisic acid (bio-ABA) was immobilized on streptavidin-coated magnetic beads. DNA aptamers against bio-ABA were selected with 7 iterative rounds of the systematic evolution of ligands by exponential enrichment method (SELEX), each round comprising incubation of the ABA-binding beads with the ssDNA sequences, DNA elution, electrophoresis, and polymerase chain reaction (PCR) amplification. The PCR product was cloned and sequenced. The binding affinity of several clones was determined using bio-ABA immobilized on streptavidin-coated plates. Aptamer 2 and aptamer 9 showed the highest binding affinity, with dissociation constants values of 0.98±0.14 μM and 0.80±0.07 μM, respectively. Aptamers 2 and 9 were also able to bind free, unmodified ABA and to discriminate between different ABA enantiomers and isomers. Our findings indicate that ssDNA aptamers can selectively bind ABA and could be used for the development of ABA quantitation assays. PMID:23971905

Highly specific and rapid immuno-fluorescent visualization and detection of E. coli O104:H4 with protein-A coated magnetic beads based LST-MUG assay.

Science.gov (United States)

Barizuddin, Syed; Balakrishnan, Baskar; Stringer, R Cody; Dweik, Majed

2015-08-01

A method combining immunomagnetic separation and fluorescent sensing was developed to detect Escherichia coli (E. coli) O104:H4. The antibody specific to E. coli O104:H4 was immobilized on protein A-coated magnetic beads. This protein-A-anti E. coli O104:H4 complex was used to bind Fluorescein IsoThioCyanate (FITC) labeled E. coli O104:H4 antigen (whole cell) on it. The goal was to achieve a fluorescently detectable protein-A-anti E. coli O104:H4-E. coli O104:H4 complex on the magnetic beads. Fluorescent microscopy was used to image the magnetic beads. The resulting fluorescence on the beads was due to the FITC labeled antigen binding on the protein-A-anti E. coli O104:H4 immobilized magnetic beads. This visually proves the antigen-antibody binding. The fluorescent imaging results were obtained in 2 h if the minimum available bacteria in the sample were at least 10(5) CFU/ml. If no fluorescence was observed on the magnetic beads during fluorescent imaging, it indicates the bacterial concentration in the sample to be too low for it to have bound to the magnetic beads and hence no detection was possible. To detect bacterial concentration less than 10(5) CFU/ml in the sample, an additional step was required for detection. The magnetic bead complex was added to the LST-MUG (lauryl sulfate tryptose-4-methylumbelliferyl-β-D-glucuronide), a signaling reporter. The E. coli O104:H4 grows in LST-MUG and releases β-glucuronidase enzyme. This enzyme cleaves the MUG substrate that produces 4-methylumbelliferone, a highly fluorescent species. This fluorescence was detected using a spectrofluorometer. The emission peak in the fluorescent spectrum was found to be at 450 nm. The lower and upper detection range for this LST-MUG assay was found to be 2.05×10(5)-4.09×10(8) CFU/ml. The results for the LST-MUG assay for concentrations below 10(5) CFU/ml were ascertained in 8h. The advantages of this technique include the specific detection of bacteria without an enrichment step and

Synthesis and characterization of Fe{sub 3}O{sub 4} nanoparticles coated with fucan polysaccharides

Energy Technology Data Exchange (ETDEWEB)

Silva, V.A.J.; Andrade, P.L. [Programa de Pós-Graduação em Ciências de Materiais, Centro de Ciências Exatas e da Natureza, Universidade Federal de Pernambuco, 50670-901 Recife-PE (Brazil); Laboratório de Imunopatologia Keizo Asami (LIKA), Universidade Federal de Pernambuco, 50670-901 Recife-PE (Brazil); Silva, M.P.C. [Laboratório de Imunopatologia Keizo Asami (LIKA), Universidade Federal de Pernambuco, 50670-901 Recife-PE (Brazil); Departamento de Bioquímica, Universidade Federal de Pernambuco, 50670-420 Recife-PE (Brazil); Bustamante D, A. [Laboratorio de Cerámicos y Nanomateriales, Facultad de Ciencias Físicas, Universidad Nacional Mayor de San Marcos, Ap. Postal 14-0149 Lima (Peru); De Los Santos Valladares, Luis [Laboratório de Imunopatologia Keizo Asami (LIKA), Universidade Federal de Pernambuco, 50670-901 Recife-PE (Brazil); Cavendish Laboratory, University of Cambridge, J.J. Thomson Avenue, Cambridge CB3 0HE (United Kingdom); Albino Aguiar, J., E-mail: albino@df.ufpe.br [Programa de Pós-Graduação em Ciências de Materiais, Centro de Ciências Exatas e da Natureza, Universidade Federal de Pernambuco, 50670-901 Recife-PE (Brazil); Departamento de Física, Universidade Federal de Pernambuco, 50670-901 Recife-PE (Brazil)

2013-10-15

In this work we report the preparation of fucan-coated magnetite (Fe{sub 3}O{sub 4}) nanoparticles by the co-precipitation method. These nanoparticles were characterized by scanning electron microscopy, transmission electron microscopy, X-ray diffraction (XRD), Fourier transform infrared (FT-IR) spectroscopy, Mössbauer spectroscopy and magnetic measurements. The nanoparticles showed quasi-spherical morphology with mean sizes around 10 nm. XRD and FT-IR confirmed the functionalization of the Fe{sub 3}O{sub 4} nanoparticles with the fucan polysaccharide. Room temperature magnetization measurements and Mössbauer spectroscopy showed that the nanoparticles exhibited superparamagnetic behavior at 300 K and the magnetic properties of the Fe{sub 3}O{sub 4} are partly screened by the coating preventing aggregation. - Highlights: • Syntheses of fucan-coated Fe{sub 3}O{sub 4} nanoparticles were made by co-precipitation method. • The efficiency of polysaccharide coated was analyzed by XRD and FT-IR. • The magnetic nanoparticles mean size was 10–20 nm. • The fucan-coated magnetite nanoparticles showed superparamagnetic behavior.

Effect of administration route and dose of streptavidin or biotin on the tumor uptake of radioactivity in intraperitoneal tumor with multistep targeting

International Nuclear Information System (INIS)

Zhang Meili; Yao Zhengsheng; Sakahara, Harumi; Saga, Tsuneo; Nakamoto, Yuji; Sato, Noriko; Zhao Songji; Nakada, Hiroshi; Yamashina, Ikuo; Konishi, Junji

1998-01-01

The effect of the administration route and dose of streptavidin or biotin on the biodistribution of radioactivity in multistep targeting was studied in nude mice bearing intraperitoneal (IP) colon cancer xenograft. The multistep targeting included a two-step method using biotinylated antibody and radiolabeled streptavidin and a three-step method with radiolabeled biotin based on the two-step method. A monoclonal antibody, MLS128, which recognizes Tn antigen on mucin, was biotinylated and injected intravenously (IV) or IP in nude mice bearing human colon cancer LS180 IP xenografts for pretargeting. In the two-step method, IP-injected streptavidin showed a higher tumor uptake and tumor-to-nontumor ratios than IV-injected streptavidin regardless of administration route of pretargeting. The tumor uptake of radiolabeled streptavidin was increased with a high dose of biotinylated antibody pretargeting, but decreased with an increasing dose of streptavidin. In the three-step targeting, IP injection also gave a higher tumor uptake of radiolabeled biotin than IV injection. In conclusion, IP administration of radiolabeled streptavidin or biotin resulted in more efficient IP tumor targeting with the multistep methods

The role of CH/π interactions in the high affinity binding of streptavidin and biotin.

Science.gov (United States)

Ozawa, Motoyasu; Ozawa, Tomonaga; Nishio, Motohiro; Ueda, Kazuyoshi

2017-08-01

The streptavidin-biotin complex has an extraordinarily high affinity (Ka: 10 15 mol -1 ) and contains one of the strongest non-covalent interactions known. This strong interaction is widely used in biological tools, including for affinity tags, detection, and immobilization of proteins. Although hydrogen bond networks and hydrophobic interactions have been proposed to explain this high affinity, the reasons for it remain poorly understood. Inspired by the deceased affinity of biotin observed for point mutations of streptavidin at tryptophan residues, we hypothesized that a CH/π interaction may also contribute to the strong interaction between streptavidin and biotin. CH/π interactions were explored and analyzed at the biotin-binding site and at the interface of the subunits by the fragment molecular orbital method (FMO) and extended applications: PIEDA and FMO4. The results show that CH/π interactions are involved in the high affinity for biotin at the binding site of streptavidin. We further suggest that the involvement of CH/π interactions at the subunit interfaces and an extended CH/π network play more critical roles in determining the high affinity, rather than involvement at the binding site. Copyright © 2017 Elsevier Inc. All rights reserved.

Se-SAD serial femtosecond crystallography datasets from selenobiotinyl-streptavidin

Science.gov (United States)

Yoon, Chun Hong; Demirci, Hasan; Sierra, Raymond G.; Dao, E. Han; Ahmadi, Radman; Aksit, Fulya; Aquila, Andrew L.; Batyuk, Alexander; Ciftci, Halilibrahim; Guillet, Serge; Hayes, Matt J.; Hayes, Brandon; Lane, Thomas J.; Liang, Meng; Lundström, Ulf; Koglin, Jason E.; Mgbam, Paul; Rao, Yashas; Rendahl, Theodore; Rodriguez, Evan; Zhang, Lindsey; Wakatsuki, Soichi; Boutet, Sébastien; Holton, James M.; Hunter, Mark S.

2017-04-01

We provide a detailed description of selenobiotinyl-streptavidin (Se-B SA) co-crystal datasets recorded using the Coherent X-ray Imaging (CXI) instrument at the Linac Coherent Light Source (LCLS) for selenium single-wavelength anomalous diffraction (Se-SAD) structure determination. Se-B SA was chosen as the model system for its high affinity between biotin and streptavidin where the sulfur atom in the biotin molecule (C10H16N2O3S) is substituted with selenium. The dataset was collected at three different transmissions (100, 50, and 10%) using a serial sample chamber setup which allows for two sample chambers, a front chamber and a back chamber, to operate simultaneously. Diffraction patterns from Se-B SA were recorded to a resolution of 1.9 Å. The dataset is publicly available through the Coherent X-ray Imaging Data Bank (CXIDB) and also on LCLS compute nodes as a resource for research and algorithm development.

Targeting of [[sup 111]In]biocytin to cultured ovarian adenocarcinoma cells using covalent monoclonal antibody -streptavidin conjugates

Energy Technology Data Exchange (ETDEWEB)

Sheldon, K.; Marks, A. (Toronto Univ., ON (Canada). Banting and Best Dept. of Medical Research); Baumal, R. (Hospital for Sick Children, Toronto, ON (Canada). Dept. of Pathology)

1992-11-01

Three monoclonal antibodies (mAb) directed against the human ovarian adenocarcinoma cell line HEY, were substituted with maleimide and covalently bonded to thiolated streptavidin. The conjugates were separated from unreacted reagents by successive affinity chromatography on protein A-Sepharose and iminobiotin columns. Purified conjugates consisted of an immunoglobulin (Ig) monomer bound to a streptavidin tetramer through a covalent bond between the Ig molecule and one of the streptavidin subunits. The conjugates were able to specifically target [[sup 111]In]biocytin to HEY cells in vitro in the presence of human serum and ascitic fluid from ovarian cancer patients. (Author).

Clinical feasibility of two-step streptavidin/111In-biotin scintigraphy in patients with suspected vertebral osteomyelitis

International Nuclear Information System (INIS)

Lazzeri, Elena; Erba, Paola A.; Volterrani, Duccio; Bottoni, Antonio; Mariani, Giuliano; Pauwels, Ernest K.J.; Manca, Mario; Bodei, Lisa; Trippi, Donatella; Cristofani, Renza; Consoli, Vincenzo; Palestro, Christopher J.

2004-01-01

Streptavidin accumulates at sites of inflammation and infection as a result of increased capillary permeability. In addition to being utilised by bacteria for their own growth, biotin forms a stable, high-affinity non-covalent complex with avidin. The objective of this investigation was to determine the diagnostic performance of two-step streptavidin/ 111 In-biotin imaging for evaluating patients with suspected vertebral osteomyelitis. We evaluated 55 consecutive patients with suspected vertebral osteomyelitis (34 women and 21 men aged 27-86 years), within 2 weeks after the onset of clinical symptoms. Thirty-two of the patients underwent magnetic resonance imaging (MRI) and 24, computed tomography (CT). DTPA-conjugated biotin was radiolabelled by incubating 500 μg of DTPA-biotin with 111 MBq of 111 In-chloride. Two-step scintigraphy was performed by first infusing 3 mg streptavidin intravenously, followed 4 h later by 111 In-biotin. Imaging was begun 60 min later. Streptavidin/ 111 In-biotin scintigraphy was positive in 32/34 patients with spinal infection (94.12% sensitivity). The study was negative in 19/21 patients without infection (95.24% specificity). The corresponding results for MRI and CT were 54.17% and 35.29% (sensitivity), and 75% and 57.14% (specificity), respectively. All statistical parameters of diagnostic performance (Youden's J index, kappa measure of agreement with correct classification, accuracy, sensitivity, specificity, positive likelihood and negative likelihood) were clearly better for streptavidin/ 111 In-biotin scintigraphy than for either MRI or CT. Streptavidin/ 111 In-biotin scintigraphy is highly sensitive and specific for detecting vertebral osteomyelitis in the first 2 weeks after the onset of clinical symptoms, and is potentially very useful for guiding clinical decisions on instituting appropriate therapy. (orig.)

Poly (dopamine) coated superparamagnetic iron oxide nanocluster for noninvasive labeling, tracking, and targeted delivery of adipose tissue-derived stem cells

Science.gov (United States)

Liao, Naishun; Wu, Ming; Pan, Fan; Lin, Jiumao; Li, Zuanfang; Zhang, Da; Wang, Yingchao; Zheng, Youshi; Peng, Jun; Liu, Xiaolong; Liu, Jingfeng

2016-01-01

Tracking and monitoring of cells in vivo after transplantation can provide crucial information for stem cell therapy. Magnetic resonance imaging (MRI) combined with contrast agents is believed to be an effective and non-invasive technique for cell tracking in living bodies. However, commercial superparamagnetic iron oxide nanoparticles (SPIONs) applied to label cells suffer from shortages such as potential toxicity, low labeling efficiency, and low contrast enhancing. Herein, the adipose tissue-derived stem cells (ADSCs) were efficiently labeled with SPIONs coated with poly (dopamine) (SPIONs cluster@PDA), without affecting their viability, proliferation, apoptosis, surface marker expression, as well as their self-renew ability and multi-differentiation potential. The labeled cells transplanted into the mice through tail intravenous injection exhibited a negative enhancement of the MRI signal in the damaged liver-induced by carbon tetrachloride, and subsequently these homed ADSCs with SPIONs cluster@PDA labeling exhibited excellent repair effects to the damaged liver. Moreover, the enhanced target-homing to tissue of interest and repair effects of SPIONs cluster@PDA-labeled ADSCs could be achieved by use of external magnetic field in the excisional skin wound mice model. Therefore, we provide a facile, safe, noninvasive and sensitive method for external magnetic field targeted delivery and MRI based tracking of transplanted cells in vivo.

Poly (dopamine) coated superparamagnetic iron oxide nanocluster for noninvasive labeling, tracking, and targeted delivery of adipose tissue-derived stem cells.

Science.gov (United States)

Liao, Naishun; Wu, Ming; Pan, Fan; Lin, Jiumao; Li, Zuanfang; Zhang, Da; Wang, Yingchao; Zheng, Youshi; Peng, Jun; Liu, Xiaolong; Liu, Jingfeng

2016-01-05

Tracking and monitoring of cells in vivo after transplantation can provide crucial information for stem cell therapy. Magnetic resonance imaging (MRI) combined with contrast agents is believed to be an effective and non-invasive technique for cell tracking in living bodies. However, commercial superparamagnetic iron oxide nanoparticles (SPIONs) applied to label cells suffer from shortages such as potential toxicity, low labeling efficiency, and low contrast enhancing. Herein, the adipose tissue-derived stem cells (ADSCs) were efficiently labeled with SPIONs coated with poly (dopamine) (SPIONs cluster@PDA), without affecting their viability, proliferation, apoptosis, surface marker expression, as well as their self-renew ability and multi-differentiation potential. The labeled cells transplanted into the mice through tail intravenous injection exhibited a negative enhancement of the MRI signal in the damaged liver-induced by carbon tetrachloride, and subsequently these homed ADSCs with SPIONs cluster@PDA labeling exhibited excellent repair effects to the damaged liver. Moreover, the enhanced target-homing to tissue of interest and repair effects of SPIONs cluster@PDA-labeled ADSCs could be achieved by use of external magnetic field in the excisional skin wound mice model. Therefore, we provide a facile, safe, noninvasive and sensitive method for external magnetic field targeted delivery and MRI based tracking of transplanted cells in vivo.

Detection of fine magnetic particles coated on a thread using an HTS-SQUID

International Nuclear Information System (INIS)

Kawagishi, K.; Itozaki, H.; Kondo, T.; Komori, K.; Koetitz, R.

2004-01-01

Polymer-coated magnetic particles, which contain superparamagnetic ferrite nanoparticles, were attached to a nylon thread of 0.35 mm in diameter and were detected by an HTS-SQUID. The length of the sample attached into the thread was within 3 mm and its interval was 30 mm. The particles were magnetized by a coil applied dc field or by a magnet of 1.4 T. The thread ran 2 mm under the SQUID with 20-100 mm/s of the rate. Signals of magnetic beads were detected and the peak-to-peak amplitude of the signals was directly proportional to the applied field and the weight of the magnetic particles. Obtained peak-to-peak amplitude for 20 ng of magnetite particles was 350 pT at 0.25 mT of applied dc field with noise of 18 pT, and estimated detection limit was 10 ng. S/N ratio was improved by the remanence measurement using the magnet and 5.8 ng of detection limit was obtained. This measurement has been proved to be promising for the continuous analysis of ultra dilute DNA solution

Synthesis and characterization of Bombesin-superparamagnetic iron oxide nanoparticles as a targeted contrast agent for imaging of breast cancer using MRI

International Nuclear Information System (INIS)

Jafari, Atefeh; Shayesteh, Saber Farjami; Salouti, Mojtaba; Heidari, Zahra; Rajabi, Ahmad Bitarafan; Boustani, Komail; Nahardani, Ali

2015-01-01

The targeted delivery of superparamagnetic iron oxide nanoparticles (SPIONs) as a contrast agent may facilitate their accumulation in cancer cells and enhance the sensitivity of MR imaging. In this study, SPIONs coated with dextran (DSPIONs) were conjugated with bombesin (BBN) to produce a targeting contrast agent for detection of breast cancer using MRI. X-ray diffraction, transmission electron microscopy, and vibrating sample magnetometer analyses indicated the formation of dextran-coated superparamagnetic iron oxide nanoparticles with an average size of 6.0 ± 0.5 nm. Fourier transform infrared spectroscopy confirmed the conjugation of the BBN with the DSPIONs. A stability study proved the high optical stability of DSPION–BBN in human blood serum. DSPION–BBN biocompatibility was confirmed by cytotoxicity evaluation. A binding study showed the targeting ability of DSPION–BBN to bind to T47D breast cancer cells overexpressing gastrin-releasing peptide (GRP) receptors. T 2 -weighted and T 2 *-weighted color map MR images were acquired. The MRI study indicated that the DSPION–BBN possessed good diagnostic ability as a GRP-specific contrast agent, with appropriate signal reduction in T 2 *-weighted color map MR images in mice with breast tumors. (paper)

Experiment list: SRX319558 [Chip-atlas[Archive

Lifescience Database Archive (English)

Full Text Available | cell type=mouse embryonic stem cells || genotype/variation=expressing control BirA || chip beads=Dynabeads... MyOne Streptavidin T1 || chip beads vendor=Invitrogen http://dbarchive.bioscienc

Experiment list: SRX319557 [Chip-atlas[Archive

Lifescience Database Archive (English)

Full Text Available se embryonic stem cells || genotype/variation=expressing Flag-bio tagged Nanog || chip beads=Dynabeads MyOne... Streptavidin T1 || chip beads vendor=Invitrogen http://dbarchive.biosciencedbc.j

Experiment list: SRX172567 [Chip-atlas[Archive

Lifescience Database Archive (English)

Full Text Available P-Seq source_name=Embryonic Stem Cell || background mouse strain=129SvJae/C57BL/6 || chip antibody=streptavidin beads... || antibody vendor/catalog=Invitrogen 656-01 Dynabeads® MyOne? Streptavidin T1 || genotype/variati

Development of simple immunoradiometric assays using avidin coupled to polystyrene beads as a common solid phase

International Nuclear Information System (INIS)

Jyotsna, N.; Singh, Y.; Chouthkanthiwar, V.; Paradkar, S.; Sivaprasad, N.

1998-01-01

In this paper, we describe the preparation and application of avidin coupled polystyrene beads as a common solid phase for use in immunoradiometric assays (IRMAs). The assay system is based on two matched commercial monoclonal antibodies, of which, the capture antibody is biotinylated using biotinamidocaproate N-hydroxysuccinimide ester and the detection antibody is radiolabeled with 125 I by conventional Chloramine-T method. Avidin was immobilized on the polystyrene beads through a primary coat of bovine serum albumin using glutaraldhyde activation method. Various factors, such as concentration of reagents, incubation time, etc. were optimised to obtain a simple assay protocol consisting of only two pipetting steps, namely, that of a mixture of the two labelled antibodies (radiolabelled and biotinylated) and of the standard or sample. The advantage of the Avidin-Biotin system is the improved sensitivity, economy of antibody and the possibility to use a common solid phase in assays for different analytes. Using the polystyrene beads along with the novel decanting device, it has been possible to achieve the convenience of the 'coated-tube' technology without the expensive automation necessary for large scale preparation of antibody coated tubes. This protocol has been successfully applied to Prolactin, LH and FSH assays. The sensitivity of the Prolactin assay is 8μIU/mL (0.3 ng/mL), that of the FSH assay is 1mIU/mL and that of the LH assay is 0.9 mIU/mL. The intra-assay and inter-assay variations were <10%. Shelf life of the avidin coupled beads was found to be about 8 months and that of the biotin labelled antibodies up to 18 months. (author)

Superparamagnetic nanoparticles for biomedical applications: Possibilities and limitations of a new drug delivery system

Energy Technology Data Exchange (ETDEWEB)

Neuberger, Tobias [Musculoskeletal Research Unit, Equine Hospital, Vetsuisse Faculty Zurich, University of Zurich, Winterthurerstr. 260, 8057 Zurich (Switzerland); Schoepf, Bernhard [Musculoskeletal Research Unit, Equine Hospital, Vetsuisse Faculty Zurich, University of Zurich, Winterthurerstr. 260, 8057 Zurich (Switzerland); Hofmann, Heinrich [Laboratory of Powder Technology, Institute of Materials, Swiss Federal Institute of Technology, EPFL, 1015 Lausanne (Switzerland); Hofmann, Margarete [MatSearch Pully, Chemin Jean Pavillard, 14, CH-1009 Pully (Switzerland); Rechenberg, Brigitte von [Musculoskeletal Research Unit, Equine Hospital, Vetsuisse Faculty Zurich, University of Zurich, Winterthurerstr. 260, 8057 Zurich (Switzerland)]. E-mail: bvonrechenberg@vetclinics.unizh.ch

2005-05-15

Nanoparticles can be used in biomedical applications, where they facilitate laboratory diagnostics, or in medical drug targeting. They are used for in vivo applications such as contrast agent for magnetic resonance imaging (MRI), for tumor therapy or cardiovascular disease. Very promising nanoparticles for these applications are superparamagnetic nanoparticles based on a core consisting of iron oxides (SPION) that can be targeted through external magnets. SPION are coated with biocompatible materials and can be functionalized with drugs, proteins or plasmids. In this review, the characteristics and applications of SPION in the biomedical sector are introduced and discussed.

Superparamagnetic nanoparticles for biomedical applications: Possibilities and limitations of a new drug delivery system

International Nuclear Information System (INIS)

Neuberger, Tobias; Schoepf, Bernhard; Hofmann, Heinrich; Hofmann, Margarete; Rechenberg, Brigitte von

2005-01-01

Nanoparticles can be used in biomedical applications, where they facilitate laboratory diagnostics, or in medical drug targeting. They are used for in vivo applications such as contrast agent for magnetic resonance imaging (MRI), for tumor therapy or cardiovascular disease. Very promising nanoparticles for these applications are superparamagnetic nanoparticles based on a core consisting of iron oxides (SPION) that can be targeted through external magnets. SPION are coated with biocompatible materials and can be functionalized with drugs, proteins or plasmids. In this review, the characteristics and applications of SPION in the biomedical sector are introduced and discussed

Aptasensor development for detection of virus in water

DEFF Research Database (Denmark)

Kirkegaard, Julie

of three types of waterborne viruses: norovirus, rotavirus and hepatitis A virus. The development of the aptasensor involved sample preparation for aptamer selection of rotavirus and hepatitis A virus, an iterative design process of the aptasensor, investigation of the surface immobilisation of aptamers...... and finally an impedimetric electrical characterisation of the sensor. The sample preparation of the rotavirus was based on purification and biotinylation of the virus to meet the requirements of the aptamer selection process. The selection process, performed by an external collaborator, was based...... on streptavidin coated magnetic bead separation, hence the needed biotinylation. It was found that the BPH linker gave the highest yield when the biotinylated rotavirus were immobilised onto the beads. The design of the viral aptasensor was determined by an iterative design process. The final chip design...

Novel multi-biotin grafted poly(lactic acid) and its self-assembling nanoparticles capable of binding to streptavidin

Science.gov (United States)

Yan, Hao; Jiang, Weimin; Zhang, Yinxing; Liu, Ying; Wang, Bin; Yang, Li; Deng, Lihong; Singh, Gurinder K; Pan, Jun

2012-01-01

Targeted drug delivery requires novel biodegradable, specific binding systems with longer circulation time. The aim of this study was to prepare biotinylated poly(lactic acid) (PLA) nanoparticles (NPs) which can meet regular requirements as well conjugate more biotins in the polymer to provide better binding with streptavidin. A biotin-graft-PLA was synthesized based on previously published biodegradable poly(ethylene glycol) (PEG)-graft-PLA, with one polymer molecule containing three PEG molecules. Newly synthesized biotin-graft-PLA had three biotins per polymer molecule, higher than the previous biotinylated PLA (≤1 biotin per polymer molecule). A PEG with a much lower molecular weight (MW ~1900) than the previous biotinylated PLA (PEG MW ≥ 3800), and thus more biocompatible, was used which supplied good nonspecific protein-resistant property compatible to PEG-graft-PLA, suggesting its possible longer stay in the bloodstream. Biotin-graft-PLA specifically bound to streptavidin and self-assembled into NPs, during which naproxen, a model small molecule (MW 230 Da) and hydrophobic drug, was encapsulated (encapsulation efficiency 51.88%). The naproxen-loaded NPs with particle size and zeta potential of 175 nm and −27.35 mV realized controlled release within 170 hours, comparable to previous studies. The biotin-graft-PLA NPs adhered approximately two-fold more on streptavidin film and on biotin film via a streptavidin arm both in static and dynamic conditions compared with PEG-graft-PLA NPs, the proven nonspecific protein-resistant NPs. The specific binding of biotin-graft-PLA NPs with streptavidin and with biotin using streptavidin arm, as well as its entrapment and controlled release for naproxen, suggest potential applications in targeted drug delivery. PMID:22334778

Experiment list: SRX319556 [Chip-atlas[Archive

Lifescience Database Archive (English)

Full Text Available ype=mouse embryonic stem cells || genotype/variation=expressing Flag-bio tagged Dax1 || chip beads=Dynabeads... MyOne Streptavidin T1 || chip beads vendor=Invitrogen http://dbarchive.bioscienc

Experiment list: SRX319553 [Chip-atlas[Archive

Lifescience Database Archive (English)

Full Text Available se embryonic stem cells || genotype/variation=expressing Flag-bio tagged Tip60 || chip beads=Dynabeads MyOne... Streptavidin T1 || chip beads vendor=Invitrogen http://dbarchive.biosciencedbc.j

Experiment list: SRX319555 [Chip-atlas[Archive

Lifescience Database Archive (English)

Full Text Available ype=mouse embryonic stem cells || genotype/variation=expressing Flag-bio tagged Dax1 || chip beads=Dynabeads... MyOne Streptavidin T1 || chip beads vendor=Invitrogen http://dbarchive.bioscienc

Experiment list: SRX319551 [Chip-atlas[Archive

Lifescience Database Archive (English)

Full Text Available use embryonic stem cells || genotype/variation=expressing Flag-bio tagged Dmap1 || chip beads=Dynabeads MyOn...e Streptavidin T1 || chip beads vendor=Invitrogen http://dbarchive.biosciencedbc.

PAMAM dendrimer-coated iron oxide nanoparticles: synthesis and characterization of different generations

International Nuclear Information System (INIS)

Khodadust, Rouhollah; Unsoy, Gozde; Yalcın, Serap; Gunduz, Gungor; Gunduz, Ufuk

2013-01-01

This study focuses on the synthesis and characterization of different generations (G 0 –G 7 ) of polyamidoamine (PAMAM) dendrimer-coated magnetic nanoparticles (DcMNPs). In this study, superparamagnetic iron oxide nanoparticles were synthesized by co-precipitation method. The synthesized nanoparticles were modified with aminopropyltrimethoxysilane for dendrimer coating. Aminosilane-modified MNPs were coated with PAMAM dendrimer. The characterization of synthesized nanoparticles was performed by X-ray diffraction, X-ray photoelectron spectroscopy (XPS), Fourier transform infrared spectroscopy (FTIR), transmission electron microscopy (TEM), dynamic light scattering, and vibrating sample magnetometry (VSM) analyses. TEM images demonstrated that the DcMNPs have monodisperse size distribution with an average particle diameter of 16 ± 5 nm. DcMNPs were found to be superparamagnetic through VSM analysis. The synthesis, aminosilane modification, and dendrimer coating of iron oxide nanoparticles were validated by FTIR and XPS analyses. Cellular internalization of nanoparticles was studied by inverted light scattering microscopy, and cytotoxicity was determined by XTT analysis. Results demonstrated that the synthesized DcMNPs, with their functional groups, symmetry perfection, size distribution, improved magnetic properties, and nontoxic characteristics could be suitable nanocarriers for targeted cancer therapy upon loading with various anticancer agents.

Experiment list: SRX172568 [Chip-atlas[Archive

Lifescience Database Archive (English)

Full Text Available us; ChIP-Seq source_name=Embryonic Stem Cell || background mouse strain=129SvJae/C57BL/6 || chip antibody=streptavidin beads... || antibody vendor/catalog=Invitrogen 656-01 Dynabeads® MyOne? Streptavidin T1 || genotype/

Structural consequences of cutting a binding loop: two circularly permuted variants of streptavidin

International Nuclear Information System (INIS)

Le Trong, Isolde; Chu, Vano; Xing, Yi; Lybrand, Terry P.; Stayton, Patrick S.; Stenkamp, Ronald E.

2013-01-01

The crystal structures of two circularly permuted streptavidins probe the role of a flexible loop in the tight binding of biotin. Molecular-dynamics calculations for one of the mutants suggests that increased fluctuations in a hydrogen bond between the protein and biotin are associated with cleavage of the binding loop. Circular permutation of streptavidin was carried out in order to investigate the role of a main-chain amide in stabilizing the high-affinity complex of the protein and biotin. Mutant proteins CP49/48 and CP50/49 were constructed to place new N-termini at residues 49 and 50 in a flexible loop involved in stabilizing the biotin complex. Crystal structures of the two mutants show that half of each loop closes over the binding site, as observed in wild-type streptavidin, while the other half adopts the open conformation found in the unliganded state. The structures are consistent with kinetic and thermodynamic data and indicate that the loop plays a role in enthalpic stabilization of the bound state via the Asn49 amide–biotin hydrogen bond. In wild-type streptavidin, the entropic penalties of immobilizing a flexible portion of the protein to enhance binding are kept to a manageable level by using a contiguous loop of medium length (six residues) which is already constrained by its anchorage to strands of the β-barrel protein. A molecular-dynamics simulation for CP50/49 shows that cleavage of the binding loop results in increased structural fluctuations for Ser45 and that these fluctuations destabilize the streptavidin–biotin complex

Oxidative stress response in neural stem cells exposed to different superparamagnetic iron oxide nanoparticles

Directory of Open Access Journals (Sweden)

Pongrac IM

2016-04-01

Full Text Available Igor M Pongrac,1 Ivan Pavičić,2 Mirta Milić,2 Lada Brkič Ahmed,1 Michal Babič,3 Daniel Horák,3 Ivana Vinković Vrček,2 Srećko Gajović1 1School of Medicine, Croatian Institute for Brain Research, University of Zagreb, 2Institute for Medical Research and Occupational Health, Zagreb, Croatia; 3Institute of Macromolecular Chemistry, Academy of Sciences of the Czech Republic, Prague, Czech Republic Abstract: Biocompatibility, safety, and risk assessments of superparamagnetic iron oxide nanoparticles (SPIONs are of the highest priority in researching their application in biomedicine. One improvement in the biological properties of SPIONs may be achieved by different functionalization and surface modifications. This study aims to investigate how a different surface functionalization of SPIONs – uncoated, coated with D-mannose, or coated with poly-L-lysine – affects biocompatibility. We sought to investigate murine neural stem cells (NSCs as important model system for regenerative medicine. To reveal the possible mechanism of toxicity of SPIONs on NSCs, levels of reactive oxygen species, intracellular glutathione, mitochondrial membrane potential, cell-membrane potential, DNA damage, and activities of SOD and GPx were examined. Even in cases where reactive oxygen species levels were significantly lowered in NSCs exposed to SPIONs, we found depleted intracellular glutathione levels, altered activities of SOD and GPx, hyperpolarization of the mitochondrial membrane, dissipated cell-membrane potential, and increased DNA damage, irrespective of the surface coating applied for SPION stabilization. Although surface coating should prevent the toxic effects of SPIONs, our results showed that all of the tested SPION types affected the NSCs similarly, indicating that mitochondrial homeostasis is their major cellular target. Despite the claimed biomedical benefits of SPIONs, the refined determination of their effects on various cellular functions

Experiment list: SRX319550 [Chip-atlas[Archive

Lifescience Database Archive (English)

Full Text Available e embryonic stem cells || genotype/variation=expressing Flag-bio tagged Myc || chip beads=Dynabeads MyOne Streptavidin T1 || chip bea...ds vendor=Invitrogen http://dbarchive.biosciencedbc.jp/k

Controlled release of D-glucose from starch granules containing 29% free D-glucose and Eudragit L100-55 as a binding and coating agent.

Science.gov (United States)

Kim, Yeon-Kye; Mukerjea, Rupendra; Robyt, John F

2010-05-27

Waxy maize starch (100% amylopectin) granules were modified by reaction of the granules with glucoamylase in a minimum amount of water to give 29% (w/w) D-glucose inside the granules [Kim, Y.-K.; Robyt, J. F. Carbohydr. Res.1999, 318, 129-134]. These granules were made into beads by dropping an ethanol slurry of starch and different amounts of Eudragit L100-55 in a constant ratio of 100:1 from a pipette onto Whatman 3MM filter paper. The starch beads were air dried and then repeatedly sprayed 0-12 times with 2.0% (w/v) Eudragit L100-55 in ethanol, with drying between each spraying, to coat the surface of the starch beads, giving different amounts of Eudragit L100-55 coating. Seven different kinds of beads, with different amounts of Eudragit L100-55 binding and coating agent, were obtained. The rates of release of D-glucose into water from the seven kinds of beads were inversely proportional to the amount of binding and coating agent. Bead type I, which was without any binding and coating gave a fast 100% release of D-glucose in 30 min. Beads II and III also gave a fast 100% release in 60 min and 90 min, respectively. Bead IV gave a near linear release of 97% D-glucose in 150 min; Bead V gave a 50% release in 120 min followed by the remaining 50% in 60 min; and Beads VI and VII gave a slow release of 10% and 4%, respectively, from 0 to 120 min, followed by a rapid 100% release from 120 to 180 min. (c) 2010 Elsevier Ltd. All rights reserved.

Chemical surface modification of glass beads for the treatment of paper machine process waters

International Nuclear Information System (INIS)

Jradi, Khalil; Daneault, Claude; Chabot, Bruno

2011-01-01

Adsorption of detrimental contaminants on a solid sorbent is proposed to remove these contaminants from process waters to increase water recycling and reduce effluent loads in the papermaking industry. A self-assembly process of attaching (covalent grafting) cationic aminosilane molecules to glass beads was investigated. The existence and the hydrolytic stability of self-assembled monolayers and multilayers were confirmed by X-Ray Photoelectron Spectroscopy and contact angle measurements. Effects of reaction time and curing on aminosilane layer structures are also discussed. The curing step after silanization seems to be crucial in the hydrophobization of the quaternary ammonium silane coated onto glass beads, and curing could affect the final chemical structure of the ammonium groups of grafted organosilane. Results indicated that modified glass beads have a strong hydrophobicity, which is attributed to the hydrophobic property of the longest carbon chain grafted onto the glass surface. Adsorption of a model contaminant (stearic acid) onto chemically modified glass beads was determined using colloidal titration. Hydrophobic interactions could be the main driving force involved between the long carbon chains of stearic acid and the carbon chains of the aminosilane layers on glass bead surfaces. Finally, self-assembly processes applied onto glass beads may have two promising applications for papermaking and self-cleaning systems.

Biotin-independent strains of Escherichia coli for enhanced streptavidin production.

Science.gov (United States)

Jeschek, Markus; Bahls, Maximilian O; Schneider, Veronika; Marlière, Philippe; Ward, Thomas R; Panke, Sven

2017-03-01

Biotin is an archetypal vitamin used as cofactor for carboxylation reactions found in all forms of life. However, biotin biosynthesis is an elaborate multi-enzymatic process and metabolically costly. Moreover, many industrially relevant organisms are incapable of biotin synthesis resulting in the requirement to supplement defined media. Here we describe the creation of biotin-independent strains of Escherichia coli and Corynebacterium glutamicum through installation of an optimized malonyl-CoA bypass, which re-routes natural fatty acid synthesis, rendering the previously essential vitamin completely obsolete. We utilize biotin-independent E. coli for the production of the high-value protein streptavidin which was hitherto restricted because of toxic effects due to biotin depletion. The engineered strain revealed significantly improved streptavidin production resulting in the highest titers and productivities reported for this protein to date. Copyright © 2017 International Metabolic Engineering Society. Published by Elsevier Inc. All rights reserved.

A Phos-tag-based magnetic-bead method for rapid and selective separation of phosphorylated biomolecules.

Science.gov (United States)

Tsunehiro, Masaya; Meki, Yuma; Matsuoka, Kanako; Kinoshita-Kikuta, Emiko; Kinoshita, Eiji; Koike, Tohru

2013-04-15

A simple and efficient method based on magnetic-bead technology has been developed for the separation of phosphorylated and nonphosphorylated low-molecular-weight biomolecules, such as nucleotides, phosphorylated amino acids, or phosphopeptides. The phosphate-binding site on the bead is an alkoxide-bridged dinuclear zinc(II) complex with 1,3-bis(pyridin-2-ylmethylamino)propan-2-olate (Phos-tag), which is linked to a hydrophilic cross-linked agarose coating on a magnetic core particle. All steps for the phosphate-affinity separation are conducted in buffers of neutral pH with 50 μL of the magnetic beads in a 1.5-mL microtube. The entire separation protocol for phosphomonoester-type compounds, from addition to elution, requires less than 12 min per sample if the buffers and the zinc(II)-bound Phos-tag magnetic beads have been prepared in advance. The phosphate-affinity magnetic beads are reusable at least 15 times without a decrease in their phosphate-binding ability and they are stable for three months in propan-2-ol. Copyright © 2013 Elsevier B.V. All rights reserved.

Potential toxicity of superparamagnetic iron oxide nanoparticles (SPION

Directory of Open Access Journals (Sweden)

Neenu Singh

2010-09-01

Full Text Available Superparamagnetic iron oxide nanoparticles (SPION are being widely used for various biomedical applications, for example, magnetic resonance imaging, targeted delivery of drugs or genes, and in hyperthermia. Although, the potential benefits of SPION are considerable, there is a distinct need to identify any potential cellular damage associated with these nanoparticles. Besides focussing on cytotoxicity, the most commonly used determinant of toxicity as a result of exposure to SPION, this review also mentions the importance of studying the subtle cellular alterations in the form of DNA damage and oxidative stress. We review current studies and discuss how SPION, with or without different surface coating, may cause cellular perturbations including modulation of actin cytoskeleton, alteration in gene expression profiles, disturbance in iron homeostasis and altered cellular responses such as activation of signalling pathways and impairment of cell cycle regulation. The importance of protein–SPION interaction and various safety considerations relating to SPION exposure are also addressed.

Tapioca starch blended alginate mucoadhesive-floating beads for intragastric delivery of Metoprolol Tartrate.

Science.gov (United States)

Biswas, Nikhil; Sahoo, Ranjan Kumar

2016-02-01

The objective of the study was to develop tapioca starch blended alginate mucoadhesive-floating beads for the intragastric delivery of Metoprolol Tartrate (MT). The beads were prepared by ionotropic gelation method using calcium chloride as crosslinker and gas forming calcium carbonate (CaCO3) as floating inducer. The alginate gel beads having 51-58% entrapped MT showed 90% release within 45 min in gastric medium (pH 1.2). Tapioca starch blending markedly improved the entrapment efficiency (88%) and sustained the release for 3-4 h. A 12% w/w HPMC coating on these beads extended the release upto 9-11 h. In vitro wash off and buoyancy test in gastric media revealed that the beads containing CaCO3 has gastric residence of more than 12 h. In vitro optimized multi-unit formulation consisting of immediate and sustained release mucoadhesive-floating beads (40:60) showed good initial release of 42% MT within 1h followed by a sustained release of over 90% for 11 h. Pharmacokinetic study performed in rabbit model showed that the relative oral bioavailability of MT after administration of oral solution, sustain release and optimized formulation was 51%, 67% and 87%, respectively. Optimized formulation showed a higher percent inhibition of isoprenaline induced heart rate in rabbits for almost 12 h. Copyright © 2015 Elsevier B.V. All rights reserved.

Surface analytical characterization of Streptavidin/poly(3-hexylthiophene) bilayers for bio-electronic applications

Science.gov (United States)

Sportelli, M. C.; Picca, R. A.; Manoli, K.; Re, M.; Pesce, E.; Tapfer, L.; Di Franco, C.; Cioffi, N.; Torsi, L.

2017-10-01

The analytical performance of bioelectronic devices is highly influenced by their fabrication methods. In particular, the final architecture of field-effect transistor biosensors combining spin-cast poly(3-hexylthiophene) (P3HT) film and a biomolecule interlayer deposited on a SiO2/Si substrate can lead to the development of highly performing sensing systems, such as for the case of streptavidin (SA) used for biotin sensing. To gain a better understanding of the quality of the interfacial area, critical is the assessment of the morphological features characteristic of the adopted biolayer deposition protocol, namely: the layer-by-layer (LbL) approach and the spin coating technique. The present study relies on a combined surface spectroscopic and morphological characterization. Specifically, X-ray photoelectron spectroscopy operated in the parallel angle-resolved mode allowed the non-destructive investigation of the in-depth chemical composition of the SA film, alone or in the presence of the P3HT overlayer. Spectroscopic data were supported and corroborated by the results obtained with a Scanning Electron and a Helium Ion microscope investigation performed on the SA layer that provided relevant information on the protein structural arrangement or on its surface morphology. Clear differences emerged between the SA layers prepared by the two approaches, with the layer-by-layer deposition resulting in a smoother and better defined bio-electronic interface. Such findings support the superior analytical performance shown by bioelectronic devices based on LbL-deposited protein layers over spin coated ones.

Enhancement of thermal conductive pathway of boron nitride coated polymethylsilsesquioxane composite.

Science.gov (United States)

Kim, Gyungbok; Ryu, Seung Han; Lee, Jun-Tae; Seong, Ki-Hun; Lee, Jae Eun; Yoon, Phil-Joong; Kim, Bum-Sung; Hussain, Manwar; Choa, Yong-Ho

2013-11-01

We report here in the fabrication of enhanced thermal conductive pathway nanocomposites of boron nitride (BN)-coated polymethylsilsesquioxane (PMSQ) composite beads using isopropyl alcohol (IPA) as a mixing medium. Exfoliated and size-reduced boron nitride particles were successfully coated on the PMSQ beads and explained by surface charge differences. A homogeneous dispersion and coating of BN on the PMSQ beads using IPA medium was confirmed by SEM. Each condition of the composite powder was carried into the stainless still mould and then hot pressed in an electrically heated hot press machine. Three-dimensional percolation networks and conductive pathways created by exfoliated BN were precisely formed in the nanocomposites. The thermal conductivity of nanocomposites was measured by multiplying specific gravity, specific heat, and thermal diffusivity, based upon the laser flash method. Densification of the composite resulted in better thermal properties. For an epoxy reinforced composite with 30 vol% BN and PMSQ, a thermal conductivity of nine times higher than that of pristine PMSQ was observed.

A giant magnetoresistance ring-sensor based microsystem for magnetic bead manipulation and detection

KAUST Repository

Gooneratne, Chinthaka P.

2011-03-28

In this paper a novel spin valvegiant magnetoresistance(GMR) ring-sensor integrated with a microstructure is proposed for concentrating, trapping, and detecting superparamagnetic beads (SPBs). Taking advantage of the fact that SPBs can be manipulated by an external magnetic field, a unique arrangement of conducting microrings is utilized to manipulate the SPBs toward the GMR sensing area in order to increase the reliability of detection. The microrings are arranged and activated in such a manner so as to enable the detection of minute concentrations of SPBs in a sample. Precise manipulation is achieved by applying current sequentially to the microrings. The fabricated ring-shaped GMR element is located underneath the innermost ring and has a magnetoresistance of approximately 5.9%. By the performed experiments it was shown that SPBs could be successfully manipulated toward the GMR sensing zone.

A giant magnetoresistance ring-sensor based microsystem for magnetic bead manipulation and detection

KAUST Repository

Gooneratne, Chinthaka P.; Giouroudi, Ioanna; Liang, Cai; Kosel, Jü rgen

2011-01-01

In this paper a novel spin valvegiant magnetoresistance(GMR) ring-sensor integrated with a microstructure is proposed for concentrating, trapping, and detecting superparamagnetic beads (SPBs). Taking advantage of the fact that SPBs can be manipulated by an external magnetic field, a unique arrangement of conducting microrings is utilized to manipulate the SPBs toward the GMR sensing area in order to increase the reliability of detection. The microrings are arranged and activated in such a manner so as to enable the detection of minute concentrations of SPBs in a sample. Precise manipulation is achieved by applying current sequentially to the microrings. The fabricated ring-shaped GMR element is located underneath the innermost ring and has a magnetoresistance of approximately 5.9%. By the performed experiments it was shown that SPBs could be successfully manipulated toward the GMR sensing zone.

SERS efficiencies of micrometric polystyrene beads coated with gold and silver nanoparticles: the effect of nanoparticle size

International Nuclear Information System (INIS)

Mir-Simon, Bernat; Morla-Folch, Judit; Pazos-Perez, Nicolas; Xie, Hai-nan; Alvarez-Puebla, Ramon A; Guerrini, Luca; Gisbert-Quilis, Patricia; Bastús, Neus G; Puntes, Víctor

2015-01-01

Rapid advances in nanofabrication techniques of reproducibly manufacturing plasmonic substrates with well-defined nanometric scale features and very large electromagnetic enhancements paved the way for the final translation of the analytical potential of surface-enhanced Raman scattering (SERS) to real applications. A vast number of different SERS substrates have been reported in the literature. Among others, discrete particles consisting of an inorganic micrometric or sub-micrometric core homogeneously coated with plasmonic nanoparticles stand out for their ease of fabrication, excellent SERS enhancing properties, long-term optical stability and remarkable experimental flexibility (manipulation, storage etc). In this article, we performed a systematic experimental study of the correlation between the size of quasi-spherical gold and silver nanoparticle and the final optical property of their corresponding assembles onto micrometric polystyrene (PS) beads. The size and composition of nanoparticles play a key role in tuning the SERS efficiency of the hybrid material at a given excitation wavelength. This study provides valuable information for the selection and optimization of the appropriate PS@NPs substrates for the desired applications. (invited article)

Efficient one-pot sonochemical synthesis of thickness-controlled silica-coated superparamagnetic iron oxide (Fe3O4/SiO2) nanospheres

Science.gov (United States)

Abbas, Mohamed; Abdel-Hamed, M. O.; Chen, Jiangang

2017-12-01

A facile and eco-friendly efficient sonochemical approach was designed for the synthesis of highly crystalline Fe3O4 and Fe3O4/SiO2 core/shell nanospheres in single reaction. The generated physical properties (shock waves, microjets, and turbulent flows) from ultrasonication as a consequence of the collapse of microbubbles and polyvinylpyrrolidone (PVP) as a chemical linker were found to play a crucial role in the successful formation of the core/shell NPs within short time than the previously reported methods. Transmission electron microscopy revealed that a uniform SiO2 shell is successfully coated over Fe3O4 nanospheres, and the thickness of the silica shell could be easily controlled in the range from 5 to 15 nm by adjusting the reaction parameters. X-ray diffraction data were employed to confirm the formation of highly crystalline and pure phase of a cubic inverse spinel structure for magnetite (Fe3O4) nanospheres. The magnetic properties of the as-synthesized Fe3O4 and Fe3O4/SiO2 core/shell nanospheres were measured at room temperature using vibrating sample magnetometer, and the results demonstrated a high magnetic moment values with superparamagnetic properties.

Experiment list: SRX319552 [Chip-atlas[Archive

Lifescience Database Archive (English)

Full Text Available embryonic stem cells || genotype/variation=expressing Flag-bio tagged E2F4 || chip beads=Dynabeads MyOne Streptavidin T1 || chip bea...ds vendor=Invitrogen http://dbarchive.biosciencedbc.jp/k

Magnetic resonance imaging of mouse islet grafts labeled with novel chitosan-coated superparamagnetic iron oxide nanoparticles.

Directory of Open Access Journals (Sweden)

Jyuhn-Huarng Juang

Full Text Available To better understand the fate of islet isografts and allografts, we utilized a magnetic resonance (MR imaging technique to monitor mouse islets labeled with a novel MR contrast agent, chitosan-coated superparamagnetic iron oxide (CSPIO nanoparticles.After being incubated with and without CSPIO (10 µg/ml, C57BL/6 mouse islets were examined under transmission electron microscope (TEM and their insulin secretion was measured. Cytotoxicity was examined in α (αTC1 and β (NIT-1 and βTC cell lines as well as islets. C57BL/6 mice were used as donors and inbred C57BL/6 and Balb/c mice were used as recipients of islet transplantation. Three hundred islets were transplanted under the left kidney capsule of each mouse and then MR was performed in the recipients periodically. At the end of study, the islet graft was removed for histology and TEM studies.After incubation of mouse islets with CSPIO (10 µg/mL, TEM showed CSPIO in endocytotic vesicles of α- and β-cells at 8 h. Incubation with CSPIO did not affect insulin secretion from islets and death rates of αTC1, NIT-1 and βTC cell lines as well as islets. After syngeneic and allogeneic transplantation, grafts of CSPIO-labeled islets were visualized on MR scans as persistent hypointense areas. At 8 weeks after syngeneic transplantation and 31 days after allogeneic transplantation, histology of CSPIO-labeled islet grafts showed colocalized insulin and iron staining in the same areas but the size of allografts decreased with time. TEM with elementary iron mapping demonstrated CSPIO distributed in the cytoplasm of islet cells, which maintained intact ultrastructure.Our results indicate that after syngeneic and allogeneic transplantation, islets labeled with CSPIO nanoparticles can be effectively and safely imaged by MR.

Superparamagnetic iron oxide nanoparticles (SPIONs) for targeted drug delivery

Science.gov (United States)

Garg, Vijayendra K.; Kuzmann, Erno; Sharma, Virender K.; Kumar, Arun; Oliveira, Aderbal C.

2016-10-01

Studies of superparamagnetic iron oxide nanoparticles (SPIONs) have been extensively carried out. Since the earlier work on Mössbauer studies on SPIONs in 1970s, many biomedical applications and their uses in innovative methods to produce new materials with improved performance have appeared. Applications of SPIONs in environmental remediation are also forthcoming. Several different methods of synthesis and coating of the magnetic particles have been described in the literature, and Mössbauer spectroscopy has been an important tool in the characterization of these materials. It is quite possible that the interpretation of the Mössbauer spectra might not be entirely correct because the possible presence of maghemite in the end product of SPIONs might not have been taken into consideration. Nanotechnology is an emerging field that covers a wide range of new technologies under development in nanoscale (1 to 100 nano meters) to produce new products and methodology.

Fluorescent nanoscale detection of biotin-streptavidin interaction using near-field scanning optical microscopy

International Nuclear Information System (INIS)

Park, Hyun Kyu; Chung, Bong Hyun; Gokarna, Anisha; Hulme, John P; Park, Hyun Gyu

2008-01-01

We describe a nanoscale strategy for detecting biotin-streptavidin binding using near-field scanning optical microscopy (NSOM) that exploits the fluorescence properties of single polydiacetylene (PDA) liposomes. NSOM is more useful to observe nanomaterials having optical properties with the help of topological information. We synthesized amine-terminated 10,12-pentacosadiynoic acid (PCDA) monomer (PCDA-NH 2 ) and used this derivatized monomer to prepare PCDA liposomes. PCDA-NH 2 liposomes were immobilized on an aldehyde-functionalized glass surface followed by photopolymerization by using a 254 nm light source. To measure the biotin-streptavidin binding, we conjugated photoactivatable biotin to immobilized PCDA-NH 2 liposomes by UV irradiation (365 nm) and subsequently allowed them to interact with streptavidin. We analyzed the fluorescence using a fluorescence scanner and observed single liposomes using NSOM. The average height and NSOM signal observed in a single liposome after binding were ∼31.3 to 8.5 ± 0.5 nm and 0.37 to 0.16 ± 0.6 kHz, respectively. This approach, which has the advantage of not requiring a fluorescent label, could prove highly beneficial for single molecule detection technology

Complementary approaches for the evaluation of biocompatibility of 90Y-labeled superparamagnetic citric acid (Fe,Er)3O4 coated nanoparticles.

Science.gov (United States)

Antic, Bratislav; Boskovic, Marko; Nikodinovic-Runic, Jasmina; Ming, Yue; Zhang, Hongguo; Bozin, Emil S; Janković, Drina; Spasojevic, Vojislav; Vranjes-Djuric, Sanja

2017-06-01

Magnetic nanoparticles (MNPs) are of immense interest for diagnostic and therapeutic applications in medicine. Design and development of new iron oxide-based MNPs for such applications is of rather limited breadth without reliable and sensitive methods to determine their levels in body tissues. Commonly used methods, such as ICP, are quite problematic, due to the inability to decipher the origin of the detected iron, i.e. whether it originates from the MNPs or endogenous from tissues and bodily fluids. One of the approaches to overcome this problem and to increase reliability of tracing MNPs is to partially substitute iron ions in the MNPs with Er. Here, we report on the development of citric acid coated (Fe,Er) 3 O 4 nanoparticles and characterization of their physico-chemical and biological properties by utilization of various complementary approaches. The synthesized MNPs had a narrow (6-7nm) size distribution, as consistently seen in atomic pair distribution function, transmission electron microscopy, and DC magnetization measurements. The particles were found to be superparamagnetic, with a pronounced maximum in measured zero-field cooled magnetization at around 90K. Reduction in saturation magnetization due to incorporation of 1.7% Er 3+ into the Fe 3 O 4 matrix was clearly observed. From the biological standpoint, citric acid coated (Fe,Er) 3 O 4 NPs were found to induce low toxicity both in human cell fibroblasts and in zebrafish (Danio rerio) embryos. Biodistribution pattern of the MNPs after intravenous administration in healthy Wistar rats was followed by the radiotracer method, revealing that 90 Y-labeled MNPs were predominantly found in liver (75.33% ID), followed by lungs (16.70% ID) and spleen (2.83% ID). Quantitative agreement with these observations was obtained by ICP-MS elemental analysis using Er as the detected tracer. Based on the favorable physical, chemical and biological characteristics, citric acid coated (Fe,Er) 3 O 4 MNPs could be

Tethered particle analysis of supercoiled circular DNA using peptide nucleic acid handles.

Science.gov (United States)

Norregaard, Kamilla; Andersson, Magnus; Nielsen, Peter Eigil; Brown, Stanley; Oddershede, Lene B

2014-09-01

This protocol describes how to monitor individual naturally supercoiled circular DNA plasmids bound via peptide nucleic acid (PNA) handles between a bead and a surface. The protocol was developed for single-molecule investigation of the dynamics of supercoiled DNA, and it allows the investigation of both the dynamics of the molecule itself and of its interactions with a regulatory protein. Two bis-PNA clamps designed to bind with extremely high affinity to predetermined homopurine sequence sites in supercoiled DNA are prepared: one conjugated with digoxigenin for attachment to an anti-digoxigenin-coated glass cover slide, and one conjugated with biotin for attachment to a submicron-sized streptavidin-coated polystyrene bead. Plasmids are constructed, purified and incubated with the PNA handles. The dynamics of the construct is analyzed by tracking the tethered bead using video microscopy: less supercoiling results in more movement, and more supercoiling results in less movement. In contrast to other single-molecule methodologies, the current methodology allows for studying DNA in its naturally supercoiled state with constant linking number and constant writhe. The protocol has potential for use in studying the influence of supercoils on the dynamics of DNA and its associated proteins, e.g., topoisomerase. The procedure takes ~4 weeks.

Metallurgical Characterization of a Weld Bead Coating Applied by the PTA Process on the D2 Tool Steel

Directory of Open Access Journals (Sweden)

Ali Tahaei

Full Text Available Abstract In this investigation, a nickel-base powder mixed with tungsten carbide particles was applied by Plasma Transferred Arc welding (PTA on the surface of the D2 cold work tool steel to improve surface quality and to extend its lifetime during applications. The Design of Experiment (DoE method was applied to obtain the appropriate combination of hardfacing parameters and to run the minimum number of tests. Current, travel speed and preheat were considered as variable parameters. These parameters are important to reach a final layer with an appropriate bead geometry accompanied with good metallurgical properties. All samples were prepared for metallurgical investigations and the effect of process parameters on the weld bead geometry was considered. For each experiment run, weld bead geometry parameters were measured including dilution, penetration and reinforcement. Microstructures and the distribution of tungsten carbide particles after welding were analyzed by Optical Microscopy (OM and Scanning Electron Microscopy (SEM equipped with an EDS microprobe. In addition, hardness tests were performed to evaluate the mechanical properties of the weld bead layers. Finally, among all the experiments, the best sample with appropriate bead geometry and microstructure was selected.

Silica-coated poly(glycidyl methacrylate-ethylene dimethacrylate) beads containing organic phase change materials

Czech Academy of Sciences Publication Activity Database

Feczkó, T.; Trif, L.; Németh, B.; Horák, Daniel

2016-01-01

Roč. 641, 10 October (2016), s. 24-28 ISSN 0040-6031 Institutional support: RVO:61389013 Keywords : porous poly(glycidyl methacrylate-ethylene dimethacrylate) beads * paraffin * cetyl alcohol Subject RIV: CD - Macromolecular Chemistry Impact factor: 2.236, year: 2016

A novel magnet focusing plate for matrix-assisted laser desorption/ionization analysis of magnetic bead-bound analytes.

Science.gov (United States)

Gode, David; Volmer, Dietrich A

2013-05-15

Magnetic beads are often used for serum profiling of peptide and protein biomarkers. In these assays, the bead-bound analytes are eluted from the beads prior to mass spectrometric analysis. This study describes a novel matrix-assisted laser desorption/ionization (MALDI) technique for direct application and focusing of magnetic beads to MALDI plates by means of dedicated micro-magnets as sample spots. Custom-made MALDI plates with magnetic focusing spots were made using small nickel-coated neodymium micro-magnets integrated into a stainless steel plate in a 16 × 24 (384) pattern. For demonstrating the proof-of-concept, commercial C-18 magnetic beads were used for the extraction of a test compound (reserpine) from aqueous solution. Experiments were conducted to study focusing abilities, the required laser energies, the influence of a matrix compound, dispensing techniques, solvent choice and the amount of magnetic beads. Dispensing the magnetic beads onto the micro-magnet sample spots resulted in immediate and strong binding to the magnetic surface. Light microscope images illustrated the homogeneous distribution of beads across the surfaces of the magnets, when the entire sample volume containing the beads was pipetted onto the surface. Subsequent MALDI analysis of the bead-bound analyte demonstrated excellent and reproducible ionization yields. The surface-assisted laser desorption/ionization (SALDI) properties of the strongly light-absorbing γ-Fe2O3-based beads resulted in similar ionization efficiencies to those obtained from experiments with an additional MALDI matrix compound. This feasibility study successfully demonstrated the magnetic focusing abilities for magnetic bead-bound analytes on a novel MALDI plate containing small micro-magnets as sample spots. One of the key advantages of this integrated approach is that no elution steps from magnetic beads were required during analyses compared with conventional bead experiments. Copyright © 2013 John Wiley

Preparation of Ca-alginate coated nZVI core shell beads for uranium (VI) removal from aqueous solution

International Nuclear Information System (INIS)

Shuhong Hu; Xiaoyan Lin; Yahui Zhang; Meiling Shi

2017-01-01

In this study, the core-shell nanoscale zero-valent iron (nZVI)@Alg-Ca beads were synthesized by coaxial electronic injection method for removal of U (VI) from aqueous solution, and characterized by SEM, EDX and XPS. The results showed that the pseudo-second-order models and the Langmuir isotherm model fitted well with the data obtained. The removal mechanism may include both physical adsorption of U (VI) on the surface or inside of core-shell nZVI@Alg-Ca beads and subsequent reduction of U (VI) to U (IV). Therefore, the core-shell nZVI@Alg-Ca beads would have an application prospect in effective removal of U (VI) contamination from aqueous solution. (author)

Fluorescence resonance energy transfer between NaYF4:Yb,Tm upconversion nanoparticles and gold nanorods: Near-infrared responsive biosensor for streptavidin

International Nuclear Information System (INIS)

Zhang, Shuang; Wang, Jing; Xu, Wen; Chen, Boting; Yu, Wei; Xu, Lin; Song, Hongwei

2014-01-01

We represent a fluorescence resonance energy transfer (FRET) system using upconversion nanoparticles (UCNPs) and the gold nanorods (GNRs) as the energy donor–acceptor pair for directly determining streptavidin in near-infrared (NIR) region. NaYF 4 :Yb,Tm UCNPs, which had a strong emission at 800 nm under 980-nm excitation, were adopted as the energy donor. The GNRs, which demonstrated strong surface plasmon absorption around 800 nm, were chosen as acceptor to quench the 800 nm emissions of the UCNPs. There had the spectral overlap between the emission of the donor nanoparticles (UCNPs) and the absorption of the acceptor nanoparticles (GNRs). This UCNP-based FRET system was then used to determine the amount of streptavidin. In this system, NaYF 4 :Yb,Tm UCNPs conjugated with biotin, while GNRs conjugated with streptavidin. When added GNRs into UCNPs, the streptavidin were preferred to bind with biotin and decreased spacing between the donor and acceptor NPs. Consequently, FRET occurred and a linear relationship between the luminescence quenching efficiency and the concentration of streptavidin was obtained. Owing to the aforementioned merits of UCNPs as an energy donor and the strong quenching ability of GNRs, satisfactory analytical performances have been acquired. -- Highlights: • NaYF4:Yb,Tm and GNRs are as NIR energy donor and quenching acceptor for FRET. • Linkage between biotin and streptavidin make the distance between the donors and the acceptors short enough for FRET. • The FRET system in this work was applicable for the detection of streptavidin. • The donor and acceptor NPs can be modified by proper molecules for other biological molecules detection

Sangadzhi Kononov, Buddhist Prayer Beads

OpenAIRE

Churyumov, Anton; Kovaeva, Bair

2016-01-01

Prayer beads have special dividers that divide the beads into 7, 21 and 33. Apart from using in prayers, the Kalmyks also keep beads as amulets that are believed to have strong energy. After prayers, old people often bless their children and grandchildren with their beads. Such beads are also kept in families from one generation to the next. Sangadzhi believes that prayer beads store inside them the energy of mantras that have been read with them. There is an interesting story about the pray...

Rapid and reagent-saving immunoassay using innovative stirring actions of magnetic beads in microreactors in the sequential injection mode.

Science.gov (United States)

Tanaka, K; Imagawa, H

2005-12-15

We developed new ELISA techniques in sequential injection analysis (SIA) mode using microreactors with content of a few microliters. We immobilized antibodies on magnetic beads 1.0mum in diameter, injected the beads into microreactors and applied rotating magnetic fields of several hundred gauss. Magnetic beads, suspended in liquid in density of approximately 10(9)-10(10) particles per millilitre, form a large number of thin rod clusters, whose length-wise axes are oriented in parallel with the magnetic field. We rotate the Nd magnets below the center of the microreactor by a tiny motor at about 2000-5000rpm. These rotating clusters remarkably accelerate the binding rate of the antibodies with antigens in the liquid. The beads are trapped around the center of the rotating magnetic field even in the flowing liquid. This newly found phenomenon enables easy bead handling in microreactors. Modification of reactor walls with selected blocking reagents was essential, because protein-coated beads often stick to the wall surface and cannot move freely. Washing steps were also shortened.

Non-immunogenic dextran-coated superparamagnetic iron oxide nanoparticles: a biocompatible, size-tunable contrast agent for magnetic resonance imaging

Directory of Open Access Journals (Sweden)

Unterweger H

2017-07-01

Full Text Available Harald Unterweger,1,* Christina Janko,1,* Marc Schwarz,2 László Dézsi,3 Rudolf Urbanics,4 Jasmin Matuszak,1 Erik Őrfi,3 Tamás Fülöp,3 Tobias Bäuerle,2 János Szebeni,3,4 Clément Journé,5 Aldo R Boccaccini,6 Christoph Alexiou,1 Stefan Lyer,1 Iwona Cicha1 1Cardiovascular Nanomedicine Unit, Section of Experimental Oncology und Nanomedicine (SEON, Else Kröner-Fresenius-Stiftung-Professorship, ENT Department, University Hospital Erlangen, Friedrich-Alexander-Universitaet Erlangen-Nuernberg, 2Preclinical Imaging Platform Erlangen (PIPE, Institute of Radiology, University Hospital Erlangen, Erlangen, Germany; 3Nanomedicine Research and Education Center, Semmelweis University, 4SeroScience Ltd., Budapest, Hungary; 5Inserm U1148, Fédération de Recherche en Imagerie Multimodalités (FRIM, X Bichat Hospital, Paris Diderot University, Paris, France; 6Institute of Biomaterials, Department of Materials Science and Engineering, University Erlangen-Nuremberg, Erlangen, Germany *These authors contributed equally to this work Abstract: Iron oxide-based contrast agents have been in clinical use for magnetic resonance imaging (MRI of lymph nodes, liver, intestines, and the cardiovascular system. Superparamagnetic iron oxide nanoparticles (SPIONs have high potential as a contrast agent for MRI, but no intravenous iron oxide-containing agents are currently approved for clinical imaging. The aim of our work was to analyze the hemocompatibility and immuno-safety of a new type of dextran-coated SPIONs (SPIONdex and to characterize these nanoparticles with ultra-high-field MRI. Key parameters related to nanoparticle hemocompatibility and immuno-safety were investigated in vitro and ex vivo. To address concerns associated with hypersensitivity reactions to injectable nanoparticulate agents, we analyzed complement activation-related pseudoallergy (CARPA upon intravenous administration of SPIONdex in a pig model. Furthermore, the size-tunability of SPIONdex and

Direct binding to antigen-coated beads refines the specificity and cross-reactivity of four monoclonal antibodies that recognize polymorphic epitopes of HLA class I molecules.

Science.gov (United States)

Hilton, H G; Parham, P

2013-04-01

Monoclonal antibodies with specificity for human leukocyte antigen (HLA) class I determinants of HLA were originally characterized using serological assays in which the targets were cells expressing three to six HLA class I variants. Because of this complexity, the specificities of the antibodies were defined indirectly by correlation. Here we use a direct binding assay, in which the targets are synthetic beads coated with 1 of 111 HLA class I variants, representing the full range of HLA-A, -B and -C variation. We studied one monoclonal antibody with monomorphic specificity (W6/32) and four with polymorphic specificity (MA2.1, PA2.1, BB7.2 and BB7.1) and compared the results with those obtained previously. W6/32 reacted with all HLA class I variants. MA2.1 not only exhibits high specificity for HLA-A*02, -B*57 and -B*58, but also exhibited cross-reactivity with HLA-A*11 and -B*15:16. At low concentration (1 µg/ml), PA2.1 and BB7.2 were both specific for HLA-A*02 and -A*69, and at high concentration (50 µg/ml) exhibited significant cross-reactions with HLA-A*68, -A*23 and -A*24. BB7.1 exhibits specificity for HLA-B*07 and -B*42, as previously described, but reacts equally well with HLA-B*81, a rare allotype defined some 16 years after the description of BB7.1. The results obtained with cell-based and bead-based assays are consistent and, in combination with amino acid sequence comparison, increase understanding of the polymorphic epitopes recognized by the MA2.1, PA2.1, BB7.2 and BB7.1 antibodies. Comparison of two overlapping but distinctive bead sets from two sources gave similar results, but the overall levels of binding were significantly different. Several weaker reactions were observed with only one of the bead sets. © 2013 John Wiley & Sons A/S.

Incorporating functionalized polyethylene glycol lipids into reprecipitated conjugated polymer nanoparticles for bioconjugation and targeted labeling of cells

Science.gov (United States)

Kandel, Prakash K.; Fernando, Lawrence P.; Ackroyd, P. Christine; Christensen, Kenneth A.

2011-03-01

We report a simple and rapid method to prepare extremely bright, functionalized, stable, and biocompatible conjugated polymer nanoparticles incorporating functionalized polyethylene glycol (PEG) lipids by reprecipitation. These nanoparticles retain the fundamental spectroscopic properties of conjugated polymer nanoparticles prepared without PEG lipid, but demonstrate greater hydrophilicity and quantum yield compared to unmodified conjugated polymer nanoparticles. The sizes of these nanoparticles, as determined by TEM, were 21-26 nm. Notably, these nanoparticles were prepared with several PEG lipid functional end groups, including biotin and carboxy moieties that can be easily conjugated to biomolecules. We have demonstrated the availability of these end groups for functionalization using the interaction of biotin PEG lipid conjugated polymer nanoparticles with streptavidin. Biotinylated PEG lipid conjugated polymer nanoparticles bound streptavidin-linked magnetic beads, while carboxy and methoxy PEG lipid modified nanoparticles did not. Similarly, biotinylated PEG lipid conjugated polymer nanoparticles bound streptavidin-coated glass slides and could be visualized as diffraction-limited spots, while nanoparticles without PEG lipid or with non-biotin PEG lipid end groups were not bound. To demonstrate that nanoparticle functionalization could be used for targeted labelling of specific cellular proteins, biotinylated PEG lipid conjugated polymer nanoparticles were bound to biotinylated anti-CD16/32 antibodies on J774A.1 cell surface receptors, using streptavidin as a linker. This work represents the first demonstration of targeted delivery of conjugated polymer nanoparticles and demonstrates the utility of these new nanoparticles for fluorescence based imaging and sensing.We report a simple and rapid method to prepare extremely bright, functionalized, stable, and biocompatible conjugated polymer nanoparticles incorporating functionalized polyethylene glycol (PEG

Investigation properties of superparamagnetic nanoparticles and magnetic field-dependent hyperthermia therapy

Science.gov (United States)

Hedayatnasab, Z.; Abnisa, F.; Daud, W. M. A. Wan

2018-03-01

The application of superparamagnetic nanoparticles as heating agents in hyperthermia therapy has made a therapeutic breakthrough in cancer treatment. The high efficiency of this magnetic hyperthermia therapy has derived from a great capability of superparamagnetic nanoparticles to generate focused heat in inaccessible tumors being effectively inactivated. The main challenges of this therapy are the improvement of the induction heating power of superparamagnetic nanoparticles and the control of the hyperthermia temperature in a secure range of 42 °C to 47 °C, at targeted area. The variation of these hyperthermia properties is principally dependent on the magnetic nanoparticles as well as the magnetic field leading to enhance the efficiency of magnetic hyperthermia therapy at targeted area and also avoid undue heating to healthy cells. The present study evaluates the magnetic hyperthermia therapy through the determination of superparamagnetic nanoparticles properties and magnetic field’ parameters.

Bead beavior in a non-continues slot die coating regime

NARCIS (Netherlands)

Langen, A.; Senes, A.; Vries, I. de; Groen, P.

2015-01-01

Sheet-to-sheet slot die coating is a crucial intermediate technique to bring organic and large area electronics (e.g. OLED and Flexible OPV) towards mass roll-to-roll productions. The coating on substrate has to be uniform within nanometers. Additional settings are added to the slot die coater for a

A two-channel detection method for autofluorescence correction and efficient on-bead screening of one-bead one-compound combinatorial libraries using the COPAS fluorescence activated bead sorting system

International Nuclear Information System (INIS)

Hintersteiner, Martin; Auer, Manfred

2013-01-01

One-bead one-compound combinatorial library beads exhibit varying levels of autofluorescence after solid phase combinatorial synthesis. Very often this causes significant problems for automated on-bead screening using TentaGel beads and fluorescently labeled target proteins. Herein, we present a method to overcome this limitation when fluorescence activated bead sorting is used as the screening method. We have equipped the COPAS bead sorting instrument with a high-speed profiling unit and developed a spectral autofluorescence correction method. The correction method is based on a simple algebraic operation using the fluorescence data from two detection channels and is applied on-the-fly in order to reliably identify hit beads by COPAS bead sorting. Our method provides a practical tool for the fast and efficient isolation of hit beads from one-bead one-compound library screens using either fluorescently labeled target proteins or biotinylated target proteins. This method makes hit bead identification easier and more reliable. It reduces false positives and eliminates the need for time-consuming pre-sorting of library beads in order to remove autofluorescent beads. (technical note)

Pitfalls using tyramide signal amplification (TSA) in the mouse gastrointestinal tract: endogenous streptavidin-binding sites lead to false positive staining.

Science.gov (United States)

Horling, L; Neuhuber, W L; Raab, M

2012-02-15

Highly sensitive immunohistochemical detection systems such as tyramide signal amplification (TSA) are widely used, since they allow using two primary antibodies raised in the same species. Most of them are based on the streptavidin-biotin-peroxidase system and include streptavidin-coupled secondary antibodies. Using TSA in cryostat-sectioned tissues of mouse esophagus, we were puzzled by negative controls with unexpected staining mostly in the ganglionic areas. This prompted us to search for the causing agent and to include also other parts of the mouse gastrointestinal tract for comparison. Streptavidin-coupled antibodies bound to endogenous binding sites yet to be characterized, which are present throughout the mouse intestines. Staining was mainly localized around neuronal cell bodies of enteric ganglia. Thus, caution is warranted when applying streptavidin-coupled antibodies in the mouse gastrointestinal tract. The use of endogenous biotin-blocking kits combined with a prolonged post-fixation time could significantly reduce unintentional staining. Copyright © 2011 Elsevier B.V. All rights reserved.

New Nanoparticles Dispersing Beads Mill with Ultra Small Beads and its Application

International Nuclear Information System (INIS)

Inkyo, M; Tahara, T; Imajyo, Y

2011-01-01

Two of the major problems related to nanoparticle dispersion with a conventional beads mill are re-agglomeration and damage to the crystalline structure of the particles. The Ultra Apex Mill was developed to solve these problems by enabling the use of ultra-small beads with a diameter of less than 0.1mm. The core of this breakthrough development is centrifugation technology which allows the use of beads as small as 0.015mm. When dispersing agglomerated nanoparticles the impulse of the small beads is very low which means there is little influence on the particles. The surface energy of the nanoparticles remains low so the properties are not likely to change. As a result, stable nanoparticle dispersions can be achieved without re-cohesion. The Ultra Apex Mill is superior to conventional beads mills that are limited to much larger bead sizes. The technology of the Ultra Apex Mill has pioneered practical applications for nanoparticles in various fields: composition materials for LCD screens, ink-jet printing, ceramic condensers and cosmetics.

Esferas (beads de alginato como agente encapsulante de óleo de croton zehntneri Pax et Hoffm ALG/Ca beads as an encapsulation agent of croton zehntneri Pax et Hoffm essential oil

Directory of Open Access Journals (Sweden)

Haroldo C. B. de Paula

2010-06-01

Full Text Available Esferas de alginato (ALG reticuladas com cálcio e revestidas com goma do cajueiro (GC foram preparadas e dopadas com óleo essencial de Croton zehntneri Pax et Hoffm (Cz, visando a seu emprego como larvicida para combate ao vetor da dengue. As esferas foram caracterizadas estruturalmente por espectroscopia de infravermelho, análise térmica, microscopia eletrônica de varredura e em relação aos seus parâmetros de transporte líquido, dopagem, embebição (Q e cinética de liberação in vitro e in vivo. Os resultados revelaram que as esferas possuem morfologia com uma alta regularidade esférica, com superfície porosa. A matriz ALG:Cz =1:1 apresentou maior dopagem e eficiência de encapsulamento e valores de Q entre 10 e 12. O perfil de liberação do óleo encapsulado apresentou-se mais prolongado nas esferas contendo goma do cajueiro, ALG-GC:Cz =1:1. A taxa de mortalidade de larvas de Stegomyia aegypti para teores de 1,81 a 4,25 mg de óleo nas esferas resultou em valores na faixa de 72 a 100%, após 24 h. A matriz polimérica mostrou-se efetiva para proteção do principio ativo até cerca de 70 dias, com perdas da ordem de 35%. A análise térmica revelou que a incorporação de Cz e GC resulta em uma matriz polimérica termicamente mais estável.Alginate beads (ALG crosslinked with calcium and coated with cashew gum (GC were prepared and loaded with an essential oil from Croton zehntneri Pax et Hoffm (Cz. The beads were characterized by FTIR, thermal properties, with respect to their size, shape, larvicide loading, swelling, and in vitro and in vivo release kinetics. Results showed that beads with ALG:Cz =1:1 relative ratio showed higher loading and encapsulation efficiency, although presenting a faster release kinetics. On the other hand, ALG beads coated with GC showed Q values from 10 to 12, representing a significant increase in the hydrophilic character of the matrix. Infrared spectroscopy and thermal analysis confirmed the

A Homogeneous Time-Resolved Fluorescence Immunoassay Method for the Measurement of Compound W.

Science.gov (United States)

Huang, Biao; Yu, Huixin; Bao, Jiandong; Zhang, Manda; Green, William L; Wu, Sing-Yung

2018-01-01

Using compound W (a 3,3'-diiodothyronine sulfate [T 2 S] immuno-crossreactive material)-specific polyclonal antibodies and homogeneous time-resolved fluorescence immunoassay assay techniques (AlphaLISA) to establish an indirect competitive compound W (ICW) quantitative detection method. Photosensitive particles (donor beads) coated with compound W or T 2 S and rabbit anti-W antibody were incubated with biotinylated goat anti-rabbit antibody. This constitutes a detection system with streptavidin-coated acceptor particle. We have optimized the test conditions and evaluated the detection performance. The sensitivity of the method was 5 pg/mL, and the detection range was 5 to 10 000 pg/mL. The intra-assay coefficient of variation averages W levels in extracts of maternal serum samples. This may have clinical application to screen congenital hypothyroidism in utero.

Fluorescence resonance energy transfer between NaYF{sub 4}:Yb,Tm upconversion nanoparticles and gold nanorods: Near-infrared responsive biosensor for streptavidin

Energy Technology Data Exchange (ETDEWEB)

Zhang, Shuang; Wang, Jing; Xu, Wen; Chen, Boting; Yu, Wei; Xu, Lin; Song, Hongwei, E-mail: songhw@jlu.edu.cn

2014-03-15

We represent a fluorescence resonance energy transfer (FRET) system using upconversion nanoparticles (UCNPs) and the gold nanorods (GNRs) as the energy donor–acceptor pair for directly determining streptavidin in near-infrared (NIR) region. NaYF{sub 4}:Yb,Tm UCNPs, which had a strong emission at 800 nm under 980-nm excitation, were adopted as the energy donor. The GNRs, which demonstrated strong surface plasmon absorption around 800 nm, were chosen as acceptor to quench the 800 nm emissions of the UCNPs. There had the spectral overlap between the emission of the donor nanoparticles (UCNPs) and the absorption of the acceptor nanoparticles (GNRs). This UCNP-based FRET system was then used to determine the amount of streptavidin. In this system, NaYF{sub 4}:Yb,Tm UCNPs conjugated with biotin, while GNRs conjugated with streptavidin. When added GNRs into UCNPs, the streptavidin were preferred to bind with biotin and decreased spacing between the donor and acceptor NPs. Consequently, FRET occurred and a linear relationship between the luminescence quenching efficiency and the concentration of streptavidin was obtained. Owing to the aforementioned merits of UCNPs as an energy donor and the strong quenching ability of GNRs, satisfactory analytical performances have been acquired. -- Highlights: • NaYF4:Yb,Tm and GNRs are as NIR energy donor and quenching acceptor for FRET. • Linkage between biotin and streptavidin make the distance between the donors and the acceptors short enough for FRET. • The FRET system in this work was applicable for the detection of streptavidin. • The donor and acceptor NPs can be modified by proper molecules for other biological molecules detection.

PCR-free detection of genetically modified organisms using magnetic capture technology and fluorescence cross-correlation spectroscopy.

Directory of Open Access Journals (Sweden)

Xiaoming Zhou

2009-11-01

Full Text Available The safety of genetically modified organisms (GMOs has attracted much attention recently. Polymerase chain reaction (PCR amplification is a common method used in the identification of GMOs. However, a major disadvantage of PCR is the potential amplification of non-target DNA, causing false-positive identification. Thus, there remains a need for a simple, reliable and ultrasensitive method to identify and quantify GMO in crops. This report is to introduce a magnetic bead-based PCR-free method for rapid detection of GMOs using dual-color fluorescence cross-correlation spectroscopy (FCCS. The cauliflower mosaic virus 35S (CaMV35S promoter commonly used in transgenic products was targeted. CaMV35S target was captured by a biotin-labeled nucleic acid probe and then purified using streptavidin-coated magnetic beads through biotin-streptavidin linkage. The purified target DNA fragment was hybridized with two nucleic acid probes labeled respectively by Rhodamine Green and Cy5 dyes. Finally, FCCS was used to detect and quantify the target DNA fragment through simultaneously detecting the fluorescence emissions from the two dyes. In our study, GMOs in genetically engineered soybeans and tomatoes were detected, using the magnetic bead-based PCR-free FCCS method. A detection limit of 50 pM GMOs target was achieved and PCR-free detection of GMOs from 5 microg genomic DNA with magnetic capture technology was accomplished. Also, the accuracy of GMO determination by the FCCS method is verified by spectrophotometry at 260 nm using PCR amplified target DNA fragment from GM tomato. The new method is rapid and effective as demonstrated in our experiments and can be easily extended to high-throughput and automatic screening format. We believe that the new magnetic bead-assisted FCCS detection technique will be a useful tool for PCR-free GMOs identification and other specific nucleic acids.

Surface modified superparamagnetic nanoparticles: Interaction with fibroblasts in primary cell culture

Energy Technology Data Exchange (ETDEWEB)

Chapa Gonzalez, Christian; Roacho Pérez, Jorge A.; Martínez Pérez, Carlos A.; Olivas Armendáriz, Imelda [Instituto de Ingeniería y Tecnología, Universidad Autónoma de Ciudad Juárez, Ave. Del Charro #610 norte, Col. Partido Romero, C.P. 32320 Cd. Juárez, Chihuahua, México (Mexico); Jimenez Vega, Florinda [Instituto de Ciencias Biomédicas, Universidad Autónoma de Ciudad Juárez, Anillo envolvente del PRONAF y Estocolmo, C.P. 32320 Cd. Juárez, Chihuahua, México (Mexico); Castrejon Parga, Karen Y. [Instituto de Ingeniería y Tecnología, Universidad Autónoma de Ciudad Juárez, Ave. Del Charro #610 norte, Col. Partido Romero, C.P. 32320 Cd. Juárez, Chihuahua, México (Mexico); Garcia Casillas, Perla E., E-mail: pegarcia@uacj.mx [Instituto de Ingeniería y Tecnología, Universidad Autónoma de Ciudad Juárez, Ave. Del Charro #610 norte, Col. Partido Romero, C.P. 32320 Cd. Juárez, Chihuahua, México (Mexico)

2014-12-05

Highlights: • An inorganic layer before an organic material shell onto MNPs improves cell viability. • The coating type and the concentration of nanoparticles directly affect cell viability. • Modified magnetite nanoparticles with organic and inorganic materials was developed. - Abstract: The development of a variety of medical applications such as drug delivery, cell labeling, and medical imaging have been possible owing to the unique features exhibited by magnetic nanoparticles. Nanoparticle–cell interaction is related to the surface aspects of nanoparticle, which may be described based on their chemistry or inorganic/organic characteristics. The coating on particle surface reduces the inter-particle interactions and provides properties such as biocompatibility. Among the coating materials used for nanoparticles employed in biomedical applications, oleic acid is one of the most utilized due to its biocompatibility. However, a major drawback with this naturally occurring fatty acid is that it is easily oxidized by cells and this reduces their performance in biomedical applications. In order to avoid the direct contact of the cell with the magnetite particle, coating with an inorganic material prior to the oleic acid shell would be effective. This would retard the magnetite dissociation thereby improve the cell viability. Here we report our investigation on the effect of surface modified magnetite nanoparticles (MNPs) on the cell viability using primary cultures incubated with those particles. We prepared magnetite nanoparticles by chemical co-precipitation method; nanoparticle surface was first modified by silanol condensation followed by chemisorption of oleic acid. All nanostructures have a particle size less than 100 nm, depending on the material coating and superparamagnetic behavior. The saturated magnetizations (M{sub s}) of the magnetite samples coated with oleic acid (MAO; 49.15 emu/g) and double shell silica-oleic acid (MSAO; 46.16 emu/g) are

Latent heat storage by silica-coated polymer beads containing organic phase change materials

Czech Academy of Sciences Publication Activity Database

Feczkó, T.; Trif, L.; Horák, Daniel

2016-01-01

Roč. 132, July (2016), s. 405-414 ISSN 0038-092X R&D Projects: GA MŠk(CZ) LH14318 Institutional support: RVO:61389013 Keywords : latent heat storage * phase change materials * porous beads by suspension polymerization Subject RIV: CD - Macromolecular Chemistry Impact factor: 4.018, year: 2016

Synthesis and characterization of cationic lipid coated magnetic nanoparticles using multiple emulsions as microreactors

Energy Technology Data Exchange (ETDEWEB)

Akbaba, Hasan; Karagöz, Uğur [Ege University, Faculty of Pharmacy, Department of Pharmaceutical Biotechnology, 35100 Izmir (Turkey); Selamet, Yusuf [Izmir Institute of Technology, Faculty of Science, Department of Physics, 35433 Izmir (Turkey); Kantarcı, A. Gülten, E-mail: gulten.kantarci@ege.edu.tr [Ege University, Faculty of Pharmacy, Department of Pharmaceutical Biotechnology, 35100 Izmir (Turkey)

2017-03-15

The aim of this study was to develop a novel iron oxide nanoparticle synthesis method with in-situ surface coating. For this purpose multiple emulsions were used as microreactors for the first time and magnetic iron oxide particles synthesized in the core of cationic solid lipid nanoparticles. DLS, SEM, TEM, VSM, Raman Spectrometer, XRD, and XPS techniques were performed for characterization of the magnetic nanoparticles. Obtained magnetic nanoparticles are superparamagnetic and no additional process was needed for surface adjustments. They are positively charged as a result of cationic lipid coating and has appropriate particle size (<30 nm) for drug or nucleic acid delivery. Structure analysis showed that magnetic core material is in the form of magnetite. Saturation magnetization value was measured as 15–17 emu g{sup −1} for lipid coated magnetic nanoparticles obtained by multiple emulsion method which is reasonably sufficient for magnetic targeting. - Highlights: • A novel iron oxide nanoparticle synthesis method with in-situ surface coating. • Combining advantages of microemulsions and multiple emulsion methods. • Multiple emulsions were used as microreactors for magnetic nanoparticle synthesis. • Superparamagnetic iron oxide particles synthesized in the core of cationic lipids. • Possible delivery systems for nucleic acids, oil soluble compounds or drugs.

Friction of N-bead macromolecules in solution: Effects of the bead-solvent interaction

International Nuclear Information System (INIS)

Uvarov, Alexander; Fritzsche, Stephan

2006-01-01

The role of the bead-solvent interaction has been studied for its influence on the dynamics of an N-bead macromolecule which is immersed into a solution. Using a Fokker-Planck equation for the phase-space distribution function of the macromolecule, we show that all the effects of the solution can be treated entirely in terms of the friction tensors which are assigned to each pair of interacting beads in the chain. For the high-density as well as for the critical solvent, the properties of these tensors are discussed in detail and are calculated by using several (realistic) choices of the bead-solvent potential. From the friction tensors, moreover, an expression for the center-of-mass friction coefficient of a (N-bead) chain macromolecule is derived. Numerical data for this coefficient for 'truncated' Lennard-Jones bead-solvent potential are compared with results from molecular dynamic simulations and from the phenomenological theoretical data as found in the literature

A more sensitive, efficient and ISO 17025 validated Magnetic Capture real time PCR method for the detection of archetypal Toxoplasma gondii strains in meat.

Science.gov (United States)

Gisbert Algaba, Ignacio; Geerts, Manon; Jennes, Malgorzata; Coucke, Wim; Opsteegh, Marieke; Cox, Eric; Dorny, Pierre; Dierick, Katelijne; De Craeye, Stéphane

2017-11-01

Toxoplasma gondii is a globally prevalent, zoonotic parasite of major importance to public health. Various indirect and direct methods can be used for the diagnosis of toxoplasmosis. Whereas serological tests are useful to prove contact with the parasite has occurred, the actual presence of the parasite in the tissues of a seropositive animal is not demonstrated. For this, a bioassay is still the reference method. As an alternative, various PCR methods have been developed, but due to the limited amount of sample that can be tested, combined with a low tissue cyst density, those have proved to be insufficiently sensitive. A major improvement of the sensitivity was achieved with magnetic capture-based DNA extraction. By combining the hybridization of specific, biotinylated probes with the capture of those probes with streptavidin-coated paramagnetic beads, T. gondii DNA can selectively be "fished out" from a large volume of meat lysate. Still, several studies showed an insufficient sensitivity compared with the mouse bioassay. Here we present a method that is more sensitive (99% limit of detection: 65.4 tachyzoites per 100g of meat), economical and reliable (ISO 17025 validated) by adding a non-competitive PCR inhibition control (co-capture of cellular r18S) and making the release of the target DNA from the streptavidin-coated paramagnetic beads UV-dependent. The presented results demonstrate the potential of the modified Magnetic Capture real time PCR as a full alternative to the mouse bioassay for the screening of various types of tissues and meat, with the additional advantage of being quantitative. Copyright © 2017 Australian Society for Parasitology. Published by Elsevier Ltd. All rights reserved.

Beautiful Beads: A Lesson in Making Beads with Friendly Clay. AMACO[R] Lesson.

Science.gov (United States)

Gamble, Harriet; Gamble, David

This lesson resource includes a brief summary of the history of bead making and historic fascination with beads as adornment. A focus on design elements, color theory, craftsmanship, and technical skill in bead making is encouraged. The plan includes lesson goals and objectives; background preparation; a glossary of terms; a list of supplies; and…

Superparamagnetic response of zinc ferrite incrusted nanoparticles

Energy Technology Data Exchange (ETDEWEB)

Lopez-Maldonado, K.L., E-mail: liliana.lopez.maldonado@gmail.com [Instituto de Ingeniería y Tecnología, Universidad Autónoma de Ciudad Juárez, Av. Del Charro 450 norte, 32310 Ciudad Juárez (Mexico); Presa, P. de la, E-mail: pmpresa@ucm.es [Instituto de Magnetismo Aplicado (UCM-ADIF-CSIC), PO Box 155, 28230 Las Rozas (Spain); Dpto. Física de Materiales, Univ. Complutense de Madrid, Madrid (Spain); Betancourt, I., E-mail: israelb@unam.mx [Departamento de Materiales Metálicos y Cerámicos, Instituto de Investigaciones en Materiales, Universidad Nacional Autónoma de México, México, D.F. 04510 (Mexico); Farias Mancilla, J.R., E-mail: rurik.farias@uacj.mx [Instituto de Ingeniería y Tecnología, Universidad Autónoma de Ciudad Juárez, Av. Del Charro 450 norte, 32310 Ciudad Juárez (Mexico); Matutes Aquino, J.A., E-mail: jose.matutes@cimav.edu.mx [Centro de Investigación en Materiales Avanzados, Miguel de Cervantes 120, 31109 Chihuahua (Mexico); Hernando, A., E-mail: antonio.hernando@externos.adif.es [Instituto de Magnetismo Aplicado (UCM-ADIF-CSIC), PO Box 155, 28230 Las Rozas (Spain); Dpto. Física de Materiales, Univ. Complutense de Madrid, Madrid (Spain); and others

2015-07-15

Highlights: • Incrusted nanoparticles are found at the surface of ZnFe{sub 2}O{sub 4} microparticles. • Magnetic contribution of nano and microparticles are analyzed by different models. • Langevin model is used to calculate the nanoparticles-superparamagnetic diameter. • Susceptibility and Langevin analysis and calculations agree with experimental data. - Abstract: Zinc ferrite is synthesized via mechano-activation, followed by thermal treatment. Spinel ZnFe{sub 2}O{sub 4} single phase is confirmed by X-ray diffraction. SEM micrographs show large particles with average particle size 〈D{sub part}〉 = 1 μm, with particles in intimate contact. However, TEM micrographs show incrusted nanocrystallites at the particles surface, with average nanocrystallite size calculated as 〈D{sub inc}〉 ≈ 5 nm. The blocking temperature at 118 K in the ZFC–FC curves indicates the presence of a superparamagnetic response which is attributable to the incrusted nanocrystallites. Moreover, the hysteresis loops show the coexistence of superpara- and paramagnetic responses. The former is observable at the low field region; meanwhile, the second one is responsible of the lack of saturation at high field region. This last behavior is related to a paramagnetic contribution coming from well-ordered crystalline microdomains. The hysteresis loops are analyzed by means of two different models. The first one is the susceptibility model used to examine separately the para- and superparamagnetic contributions. The fittings with the theoretical model confirm the presence of the above mentioned magnetic contributions. Finally, using the Langevin-based model, the average superparamagnetic diameter 〈D{sub SPM}〉 is calculated. The obtained value 〈D{sub SPM}〉 = 4.7 nm (∼5 nm) is consistent with the average nanocrystallite size observed by TEM.

Size-controlled synthesis of superparamagnetic iron oxide nanoparticles and their surface coating by gold for biomedical applications

Energy Technology Data Exchange (ETDEWEB)

Maleki, H. [Institute for Nanoscience and Nanotechnology, Sharif University of Technology, Tehran (Iran, Islamic Republic of); Simchi, A., E-mail: simchi@sharif.edu [Institute for Nanoscience and Nanotechnology, Sharif University of Technology, Tehran (Iran, Islamic Republic of); Department of Material Science and Engineering, Sharif University of Technology, Tehran (Iran, Islamic Republic of); Imani, M. [Novel Drug Delivery Systems Department, Iran Polymer and Petrochemical Institute, Tehran (Iran, Islamic Republic of); Costa, B.F.O. [CEMDRX, Department of Physics, University of Coimbra, P-3004-516 Coimbra (Portugal)

2012-11-15

The size mono-dispersity, saturation magnetization, and surface chemistry of magnetic nanoparticles (NPs) are recognized as critical factors for efficient biomedical applications. Here, we performed modified water-in-oil inverse nano-emulsion procedure for preparation of stable colloidal superparamagnetic iron oxide NPs (SPIONs) with high saturation magnetization. To achieve mono-dispersed SPIONs, optimization process was probed on several important factors including molar ratio of iron salts [Fe{sup 3+} and Fe{sup 2+}], the concentration of ammonium hydroxide as reducing agent, and molar ratio of water to surfactant. The biocompatibility of the obtained NPs, at various concentrations, was evaluated via MTT (3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide) assay and the results showed that the NPs were non-toxic at concentrations <0.1 mg/mL. Surface functionalization was performed by conformal coating of the NPs with a thin shell of gold ({approx}4 nm) through chemical reduction of attached gold salts at the surface of the SPIONs. The Fe{sub 3}O{sub 4} core/Au shell particles demonstrate strong plasmon resonance absorption and can be separated from solution using an external magnetic field. Experimental data from both physical and chemical determinations of the changes in particle size, surface plasmon resonance optical band, phase components, core-shell surface composition, and magnetic properties have confirmed the formation of the mono-dispersed core-shell nanostructure. - Highlights: Black-Right-Pointing-Pointer Increasing the concentration of iron salts, cubic-shape SPION NPs were formed. The magnetic saturation of the SPIONs was also increased. Black-Right-Pointing-Pointer The concentration of reducing agent exhibited marginal effect on the size of SPIONs but influenced the crystallinity of the NPs. A lower magnetic saturation was obtained at higher NH{sub 4}OH concentrations. Black-Right-Pointing-Pointer Mono-dispersed SPIONs can be prepared

Glass bead cultivation of fungi

DEFF Research Database (Denmark)

Droce, Aida; Sørensen, Jens Laurids; Giese, H.

2013-01-01

Production of bioactive compounds and enzymes from filamentous fungi is highly dependent on cultivation conditions. Here we present an easy way to cultivate filamentous fungi on glass beads that allow complete control of nutrient supply. Secondary metabolite production in Fusarium graminearum...... and Fusarium solani cultivated on agar plates, in shaking liquid culture or on glass beads was compared. Agar plate culture and glass bead cultivation yielded comparable results while liquid culture had lower production of secondary metabolites. RNA extraction from glass beads and liquid cultures was easier...... to specific nutrient factors. •Fungal growth on glass beads eases and improves fungal RNA extraction....

PEGylated superparamagnetic iron oxide nanoparticles labeled with 68Ga as a PET/MRI contrast agent. A biodistribution study

International Nuclear Information System (INIS)

Afsaneh Lahooti; Gruttner, Cordula; Parham Geramifar; Hassan Yousefnia

2017-01-01

The purpose of this study is to evaluate the biodistribution of polyethylene glycol (PEG) coated superparamagnetic iron oxide nanoparticles radiolabeled with 68 Ga in normal mice after intravenous administration of this probe. Three mice were sacrificed at specific time intervals. The biodistribution data revealed high uptake by liver and spleen (60.62 and 12.65 %ID/g at 120 min post injection for liver and spleen, respectively). The clearance of other organs was fast. These results suggest that 68 Ga-PEG-SPIONs has magnificent capabilities for applying in (PET-MRI) as a theranostic agent for detection of liver and spleen malignancies. (author)

Heterogeneous time-resolved electrochemiluminoimmunoassay of thyroid stimulating hormone with magnetic beads at oxide-covered aluminum electrode

Energy Technology Data Exchange (ETDEWEB)

Ala-Kleme, Timo, E-mail: timo.ala-kleme@utu.fi

2017-06-15

The heterogeneous immunoassay of thyroid stimulating hormone (TSH) was detected by the time-resolved cathodic electrochemiluminescence immunoassay (tr-CECLIA) method using magnetic beads as a mobile support. The magnetic beads coated with sandwich type TSH immunoassay were captured with a magnet for washing and detection processes. The time-resolved cathodic electrochemiluminescence (tr-CECL) signal of Tb(III) chelate label was generated by cathodic pulse polarization in the aluminum working electrode and platinum counter electrode system. The detection method causes injection of high energy electrons into the aqueous solution near the aluminum electrode and creates rigid simultaneous oxidative and reductive conditions that excitate the Tb(III) chelate used as a label luminophore in the heterogeneous sandwich type immunoassay of TSH in the surface of the magnetic beads. The limit of detection of the method was about 50 mIU L{sup −1}. The precision of it was noticed to be good; the coefficient variation percentage was realized to be lower than 10 %. Unfortunately the limit of detection is not good enough for determination of analyte levels of very low concentrations for instance TSH in body fluids. The possible application areas of the method are in highly sophisticated micro or nano fluidic detection and sensor systems where the aspiration level of the analyte detection limit is not very high and the mobility and manageability and the large surface area of the magnetic beads can be utilized efficiently in separating, washing, moving, coating and detecting processes. In the case of tr-CECLIA the presented method makes possible to use multipurpose working electrodes instead of disposable ones.

Experimental study of the biological properties of 188Re-Hepama-1 biologic superparamagnetic nanoparticles

International Nuclear Information System (INIS)

Feng Yanlin; Tan Jiaju; Sun Jing; Wen Guanghua; Wu Xiaolian; Liang Sheng; Xia Jiaoyun

2007-01-01

Objective: To investigate a new biologic-superparamagnetic nanoparticles's characteristics of immunological activity, biological distributing in vivo, targeting and inhibiting tumor effect. Methods: The experimental group 188 Re-Hepama-l-superparamagnetic nanoparticles, and control groups, including 188 ReO 4 - , 188 Re-Hepama-1, and 188 Re-superparamagnetic nanoparticles, were set up. The distributions were measured after injection 4 h and 24 h by caudal vein of Kuming mice. The magnetic targeting experiments in vivo were clone with and without magnetic field in liver after injection in New Zealand rabbit. The inhibiting tumor effect on hepatic cancer cell lines SMMC-7721 of the above four 188 Re labeled products were measured by mono nuclear cell direct cytotoxicity assay method. Results: After injection 4 h and 24 h by vein, the liver taking was highest in group 188 Re-Hepama-l-superparamagnetic nanoparticles. The radiative activity in liver in magnetism zoo was higher than in non magnetism zoo in 188 Re- Hepama-1-superparamagnetic nanoparticles after applying magnetic field in left lobe of liver, and the ratio of in magnetism zoo to non magnetism zoo was 1.87. And the half effective inhibition radioactive concentrations (IC 50 ) in 188 Re-Hepama-l-superparamagnetic nanoparticles was one forth of 188 ReO 4 - . Conclusion: 188 Re- Hepama-l-superparamagnetic nanoparticles showed its fine stability in intro, good immunological activity and significant liver target. (authors)

Current status of superparamagnetic iron oxide contrast agents for liver magnetic resonance imaging.

Science.gov (United States)

Wang, Yi-Xiang J

2015-12-21

Five types of superparamagnetic iron oxide (SPIO), i.e. Ferumoxides (Feridex(®) IV, Berlex Laboratories), Ferucarbotran (Resovist(®), Bayer Healthcare), Ferumoxtran-10 (AMI-227 or Code-7227, Combidex(®), AMAG Pharma; Sinerem(®), Guerbet), NC100150 (Clariscan(®), Nycomed,) and (VSOP C184, Ferropharm) have been designed and clinically tested as magnetic resonance contrast agents. However, until now Resovist(®) is current available in only a few countries. The other four agents have been stopped for further development or withdrawn from the market. Another SPIO agent Ferumoxytol (Feraheme(®)) is approved for the treatment of iron deficiency in adult chronic kidney disease patients. Ferumoxytol is comprised of iron oxide particles surrounded by a carbohydrate coat, and it is being explored as a potential imaging approach for evaluating lymph nodes and certain liver tumors.

A magnetic biosensor system for detection of E. coli

KAUST Repository

Li, Fuquan

2013-07-01

This work describes a device for detecting E. coli bacteria by manipulating superparamagnetic beads to a sensing area and immobilizing them in a trapping well. The trapping well replaces the biochemical immobilization layer, which is commonly used in magnetic biosensor systems. A concept exploiting the volume difference between bare magnetic beads and magnetic bead-bioanalyte compounds is utilized to detect E. coli bacteria. Trapped beads are detected by the help of a tunnel magneto-resistive sensor. Frequency modulation is employed, in order to increase the signal-to-noise ratio, enabling the detection of individual superparamagnetic beads of 2.8 μm in diameter. Replacing the biochemical immobilization layer by the trapping well greatly simplifies the detection process. After applying the mixture of E. coli and magnetic beads to the biosensor system, bacteria detection is achieved in a single step, within a few minutes. © 2013 IEEE.

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.

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

International Nuclear Information System (INIS)

Awsiuk, K.; Rysz, J.; Petrou, P.; Budkowski, A.; Bernasik, A.; Kakabakos, S.; Marzec, M.M.; Raptis, I.

2014-01-01

To immobilize effectively oligonucleotide probes on SiO 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 2 ) and second (1.31(±0.22) mg/m 2 ) approach, as compared to lower surface density resulting from the third (0.58(±0.20) mg/m 2 ) and fourth (0.41(±0.11) mg/m 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.

Preparation of gold nanoparticle dimers via streptavidin-induced interlinking

International Nuclear Information System (INIS)

Zon, Vera B.; Sachsenhauser, Matthias; Rant, Ulrich

2013-01-01

There is great interest in establishing efficient means of organizing nanoparticles into complex structures, especially in fields like nano-optical devices. One of the demonstrated routes uses biomolecular scaffolds, like the streptavidin–biotin system, to deterministically separate and structure particle complexes. However, controlled formation of streptavidin-linked nanoparticle dimers or trimers is challenging, and large aggregates are often formed under conditions that are difficult to regulate. Here, we studied the aggregates and interlinking kinetics of biotin-functionalized 20 nm gold nanoparticles in the presence of the interlinking protein, streptavidin. We found two different protein-linker concentration regions where small stable particle aggregates are formed: when the protein and nanoparticle concentrations are similar and when the protein to nanoparticle concentration ratio exceeds intermediate concentrations (10:1–100:1) that promote precipitation of large aggregates. We attribute this behavior to the limited availability of free-linker molecules and the limited availability of free ligand (biotin) on the particle surface for low and high protein concentrations, respectively. Furthermore, we show that the product can be additionally enriched up to 25 % through either centrifugation in sucrose or size-exclusion chromatography. These results provide additional understanding into the assembly of ligand-functionalized nanoparticles with water-soluble linkers and provide a facile way to produce well-defined small aggregates for potential use in, for instance, surface-enhanced spectroscopy

Confocal nanoscanning, bead picking (CONA): PickoScreen microscopes for automated and quantitative screening of one-bead one-compound libraries.

Science.gov (United States)

Hintersteiner, Martin; Buehler, Christof; Uhl, Volker; Schmied, Mario; Müller, Jürgen; Kottig, Karsten; Auer, Manfred

2009-01-01

Solid phase combinatorial chemistry provides fast and cost-effective access to large bead based libraries with compound numbers easily exceeding tens of thousands of compounds. Incubating one-bead one-compound library beads with fluorescently labeled target proteins and identifying and isolating the beads which contain a bound target protein, potentially represents one of the most powerful generic primary high throughput screening formats. On-bead screening (OBS) based on this detection principle can be carried out with limited automation. Often hit bead detection, i.e. recognizing beads with a fluorescently labeled protein bound to the compound on the bead, relies on eye-inspection under a wide-field microscope. Using low resolution detection techniques, the identification of hit beads and their ranking is limited by a low fluorescence signal intensity and varying levels of the library beads' autofluorescence. To exploit the full potential of an OBS process, reliable methods for both automated quantitative detection of hit beads and their subsequent isolation are needed. In a joint collaborative effort with Evotec Technologies (now Perkin-Elmer Cellular Technologies Germany GmbH), we have built two confocal bead scanner and picker platforms PS02 and a high-speed variant PS04 dedicated to automated high resolution OBS. The PS0X instruments combine fully automated confocal large area scanning of a bead monolayer at the bottom of standard MTP plates with semiautomated isolation of individual hit beads via hydraulic-driven picker capillaries. The quantification of fluorescence intensities with high spatial resolution in the equatorial plane of each bead allows for a reliable discrimination between entirely bright autofluorescent beads and real hit beads which exhibit an increased fluorescence signal at the outer few micrometers of the bead. The achieved screening speed of up to 200,000 bead assayed in less than 7 h and the picking time of approximately 1 bead

Purification of CD47-streptavidin fusion protein from bacterial lysate using biotin-agarose affinity chromatography.

Science.gov (United States)

Salehi, Nasrin; Peng, Ching-An

2016-07-08

CD47 is a widely expressed transmembrane glycoprotein that modulates the activity of a plethora of immune cells via its extracellular domain. Therefore, CD47 plays important roles in the regulation of immune responses and may serve as targets for the development of immunotherapeutic agents. To make sure CD47 functionality is intact under the process of protein conjugation, CD47-streptavidin fusion protein was expressed and purified because it can easily bind to biotin-tagged materials via the unique biotin-streptavidin affinity. In this study, gene sequences of CD47 extracellular domain (CD47ECD) and core streptavidin (coreSA) with a total 834 bp were inserted into pET20b plasmid to construct recombinant plasmid encoding CD47-SA fusion gene. After bacteria transformation, the CD47-SA fusion protein was expressed by isopropyl-β-d-thiogalactopyranoside (IPTG) induction. The collected bacteria lysate was loaded on biotinylated agarose to proceed the purification of CD47-SA fusion protein. Due to the unexpected high affinity between biotin and coreSA, standard washing and elution approaches (e.g., varying pH, using biotin, and applying guanidine hydrochloride) reported for biotin-streptavidin affinity chromatography were not able to separate the target fusion protein. Instead, using low concentration of the non-ionic detergent Triton X-100 followed with alkaline buffer could efficiently weaken the binding between biotin and coreSA, thereby eluting out CD47-SA fusion protein from the biotin agarose column. The purified CD47-SA fusion protein was further characterized by molecular biology methods and its antiphagocytic functionality was confirmed by the phagocytosis assay. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:949-958, 2016. © 2016 American Institute of Chemical Engineers.

Preparation of magnetic nanoparticles embedded in polystyrene microspheres

International Nuclear Information System (INIS)

Nguyen Hoang Hai; Nguyen Hoang Luong; Nguyen Chau; Ngo Quy Tai

2009-01-01

Superparamagnetic particles are widely used for biological applications such as cell separation. The size of the particles is normally in the range of 10 - 20 nm which is much smaller than the size of a cell. Therefore small particles create small force which is not strong enough to separate the cells from solution. Superparamagnetic nanoparticles embedded in Polystyrene microspheres (magnetic beads) are very useful for cell separation. Magnetic beads have been prepared by solvent evaporation of an emulsion. The beads with size of 0.2 μm - 1.0 μm have a saturation magnetization of 10 - 25 emu/g. The change of the amount of surfactants, volatile solvent, magnetic particles resulted to the change of size, magnetic properties of the magnetic beads.

Streptavidin and its biotin complex at atomic resolution

International Nuclear Information System (INIS)

Le Trong, Isolde; Wang, Zhizhi; Hyre, David E.; Lybrand, Terry P.; Stayton, Patrick S.; Stenkamp, Ronald E.

2011-01-01

Analysis of atomic resolution crystal structures of wild-type streptavidin (1.03 Å) and its biotin complex (0.95 Å) indicate the range of conformational states taken on by this protein in the solid state. Most of the structural variation is found in the polypeptide loops between the strands in this β-sandwich protein. Atomic resolution crystallographic studies of streptavidin and its biotin complex have been carried out at 1.03 and 0.95 Å, respectively. The wild-type protein crystallized with a tetramer in the asymmetric unit, while the crystals of the biotin complex contained two subunits in the asymmetric unit. Comparison of the six subunits shows the various ways in which the protein accommodates ligand binding and different crystal-packing environments. Conformational variation is found in each of the polypeptide loops connecting the eight strands in the β-sandwich subunit, but the largest differences are found in the flexible binding loop (residues 45–52). In three of the unliganded subunits the loop is in an ‘open’ conformation, while in the two subunits binding biotin, as well as in one of the unliganded subunits, this loop ‘closes’ over the biotin–binding site. The ‘closed’ loop contributes to the protein’s high affinity for biotin. Analysis of the anisotropic displacement parameters included in the crystallographic models is consistent with the variation found in the loop structures and the view that the dynamic nature of the protein structure contributes to the ability of the protein to bind biotin so tightly

Fluorescence Enhancement of Fluorescent Unnatural Streptavidin by Binding of a Biotin Analogue with Spacer Tail and Its Application to Biotin Sensing

Directory of Open Access Journals (Sweden)

Xianwei Zhu

2014-01-01

Full Text Available We designed a novel molecular biosensing system for the detection of biotin, an important vitamin by the combination of fluorescent unnatural streptavidin with a commercialized biotin-(AC52-hydrazide. A fluorescent unnatural amino acid, BODIPY-FL-aminophenylalanine (BFLAF, was position-specifically incorporated into Trp120 of streptavidin by four-base codon method. Fluorescence of the Trp120BFLAF mutant streptavidin was enhanced by the addition of biotin-(AC52-hydrazide with the concentration dependent, whereas fluorescence enhancement was not observed at all by the addition of natural biotin. It was considered that the spacer tail of biotin-(AC52-hydrazide may disturb the fluorescence quenching of the Trp120BFLAF by Trp79 and Trp108 of the neighbor subunit. Therefore, biotin sensing was carried out by the competitive binding reaction of biotin-(AC52-hydrazide and natural biotin to the fluorescent mutant streptavidin. The fluorescence intensity decreased by increasing free biotin concentration. The result suggested that molecular biosensor for small ligand could be successfully designed by the pair of fluorescent mutant binding protein and ligand analogue.

Optical enhancement of phosphor-converted wLEDs using glass beads

Science.gov (United States)

Güner, Tuğrul; Şentürk, Ufuk; Demir, Mustafa M.

2017-10-01

YAG:Ce3+ is a yellow-source compound commonly used in phosphor conversion layers for direct coating or remote phosphor configurations in LED illumination. This material, however, suffers from a high correlated color temperature, and low color-rendering index due to its deficiency in the red spectrum. In this study, glass beads (GB) with an average particle diameter of 10 μm were introduced to the conversion layer of a YAG:Ce3+ particulate-filled polydimethylsiloxane matrix composite structure and found to improve the optical features of the resulting composite.

Polymerase-free measurement of microRNA-122 with single base specificity using single molecule arrays: Detection of drug-induced liver injury.

Directory of Open Access Journals (Sweden)

David M Rissin

Full Text Available We have developed a single probe method for detecting microRNA from human serum using single molecule arrays, with sequence specificity down to a single base, and without the use of amplification by polymerases. An abasic peptide nucleic acid (PNA probe-containing a reactive amine instead of a nucleotide at a specific position in the sequence-for detecting a microRNA was conjugated to superparamagnetic beads. These beads were incubated with a sample containing microRNA, a biotinylated reactive nucleobase-containing an aldehyde group-that was complementary to the missing base in the probe sequence, and a reducing agent. When a target molecule with an exact match in sequence hybridized to the capture probe, the reactive nucleobase was covalently attached to the backbone of the probe by a dynamic covalent chemical reaction. Single molecules of the biotin-labeled probe were then labeled with streptavidin-β-galactosidase (SβG, the beads were resuspended in a fluorogenic enzyme substrate, loaded into an array of femtoliter wells, and sealed with oil. The array was imaged fluorescently to determine which beads were associated with single enzymes, and the average number of enzymes per bead was determined. The assay had a limit of detection of 500 fM, approximately 500 times more sensitive than a corresponding analog bead-based assay, with target specificity down to a single base mis-match. This assay was used to measure microRNA-122 (miR-122-an established biomarker of liver toxicity-extracted from the serum of patients who had acute liver injury due to acetaminophen, and control healthy patients. All patients with liver injury had higher levels of miR-122 in their serum compared to controls, and the concentrations measured correlated well with those determined using RT-qPCR. This approach allows rapid quantification of circulating microRNA with single-based specificity and a limit of quantification suitable for clinical use.

Ferroferric oxide/polystyrene (Fe3O4/PS superparamagnetic nanocomposite via facile in situ bulk radical polymerization

Directory of Open Access Journals (Sweden)

2010-03-01

Full Text Available Organo-modified ferroferric oxide superparamagnetic nanoparticles, synthesized by the coprecipitation of superparamagnetic nanoparticles in presence of oleic acid (OA, were incorporated in polystyrene (PS by the facile in situ bulk radical polymerization by using 2,2-azobisisobutyronitrile (AIBN as initiator. The transmission electron microscopy (TEM analysis of the resultant uniform ferroferric oxide/polystyrene superparamagnetic nanocomposite (Fe3O4/PS showed that the superparamagnetic nanoparticles had been dispersed homogeneously in the polymer matrix due to the surface grafted polystyrene, confirmed by Fourier transform infrared (FT-IR spectroscopy and thermogravimetric analysis (TGA. The superparamagnetic property of the Fe3O4/PS nanocomposite was testified by the vibrating sample magnetometer (VSM analysis. The strategy developed is expected to be applied for the large-scale industrial manufacturing of the superparamagnetic polymer nanocomposite.

Catalase coupled gold nanoparticles: Comparison between carbodiimide and biotin-streptavidin methods

Science.gov (United States)

Chirra, Hariharasudhan D.; Sexton, Travis; Biswal, Dipti; Hersh, Louis B.; Hilt, J. Zach

2011-01-01

The use of proteins for therapeutic applications requires the protein to maintain sufficient activity for the period of in vivo treatment. Many proteins exhibit a short half-life in vivo and, thus, require delivery systems for them to be applied as therapeutics. The relative biocompatibility and the ability to form functionalized bioconjugates via simple chemistry make gold nanoparticles excellent candidates as protein delivery systems. Herein, two protocols for coupling proteins to gold nanoparticles were compared. In the first, the strong biomolecular binding between biotin and streptavidin was used to couple catalase to the surface of gold nanoparticles. In the second protocol, the formation of an amide bond between carboxylic acid coated gold nanoparticles and free surface amines of catalase using carbodiimide chemistry was performed. The stability and kinetics of the different steps involved in these protocols were studied using UV-Visible spectroscopy, dynamic light scattering, and transmission electron microscopy. The addition of mercaptoundecanoic acid in conjugation with (N-(6-(biotinamido)hexyl)-3′-(2′-pyridyldithio)-propionamide increased the stability of biotinylated gold nanoparticles. Although the carbodiimide chemistry based bioconjugation approach exhibited a decrease in catalase activity, the carbodiimide chemistry based bioconjugation approach resulted in more active catalase per gold nanoparticle compared to that of mercaptoundecanoic acid stabilized biotinylated gold nanoparticles. Both coupling protocols resulted in gold nanoparticles loaded with active catalase. Thus, these gold nanoparticle systems and coupling protocols represent promising methods for the application of gold nanoparticles for protein delivery. PMID:21232642

Targeting experimental orthotopic glioblastoma with chitosan-based superparamagnetic iron oxide nanoparticles (CS-DX-SPIONs).

Science.gov (United States)

Shevtsov, Maxim; Nikolaev, Boris; Marchenko, Yaroslav; Yakovleva, Ludmila; Skvortsov, Nikita; Mazur, Anton; Tolstoy, Peter; Ryzhov, Vyacheslav; Multhoff, Gabriele

2018-01-01

Glioblastoma is the most devastating primary brain tumor of the central nervous system in adults. Magnetic nanocarriers may help not only for a targeted delivery of chemotherapeutic agents into the tumor site but also provide contrast enhancing properties for diagnostics using magnetic resonance imaging (MRI). Synthesized hybrid chitosan-dextran superparamagnetic nanoparticles (CS-DX-SPIONs) were characterized using transmission electron microscopy (TEM) and relaxometry studies. Nonlinear magnetic response measurements were employed for confirming the superparamagnetic state of particles. Following in vitro analysis of nanoparticles cellular uptake tumor targeting was assessed in the model of the orthotopic glioma in rodents. CS-DX-SPIONs nanoparticles showed a uniform diameter of 55 nm under TEM and superparamagentic characteristics as determined by T 1 (spin-lattice relaxation time) and T 2 (spin-spin relaxation time) proton relaxation times. Application of the chitosan increased the charge from +8.9 to +19.3 mV of the dextran-based SPIONs. The nonlinear magnetic response at second harmonic of CS-DX-SPIONs following the slow change of stationary magnetic fields with very low hysteresis evidenced superparamagnetic state of particles at ambient temperatures. Confocal microscopy and flow cytometry studies showed an enhanced internalization of the chitosan-based nanoparticles in U87, C6 glioma and HeLa cells as compared to dextran-coated particles. Cytotoxicity assay demonstrated acceptable toxicity profile of the synthesized nanoparticles up to a concentration of 10 μg/ml. Intravenously administered CS-DX-SPIONs in orthotopic C6 gliomas in rats accumulated in the tumor site as shown by high-resolution MRI (11.0 T). Retention of nanoparticles resulted in a significant contrast enhancement of the tumor image that was accompanied with a dramatic drop in T 2 values ( P chitosan-dextran magnetic particles demonstrated high MR contrast enhancing properties for the

Poly(ethylene oxide)-block-poly(glutamic acid) coated maghemite nanoparticles: in vitro characterization and in vivo behaviour

International Nuclear Information System (INIS)

Kaufner, L; Cartier, R; Wuestneck, R; Fichtner, I; Pietschmann, S; Bruhn, H; Schuett, D; Thuenemann, A F; Pison, U

2007-01-01

Positively charged superparamagnetic iron oxide (SPIO) particles of maghemite were prepared in aqueous solution and subsequently stabilized with poly(ethylene oxide)-block-poly(glutamic acid) (PEO-PGA) at a hydrodynamic diameter of 60 nm. Depending on the amount of PEO-PGA used, this is accompanied by a switching of their zeta potentials from positive to negative charge (-33 mV). As a prerequisite for in vivo testing, the PEO-PGA coated maghemite nanoparticles were evaluated to be colloidally stable in water and in physiological salt solution for longer than six months as well in various buffer systems under physiological pH and salt conditions (AFM, dynamic light scattering). We excluded toxic effects of the PEO-PGA coated maghemite nanoparticles. We demonstrated by in vivo MR-imaging and 111 In measurements a biodistribution of the nanoparticles into the liver comparable to carboxydextran coated superparamagnetic iron oxide nanoparticles (Resovist[reg]) as a reference nanoscaled MRI contrast medium. This was enforced by a detailed visualization of our nanoparticles by electron microscopy of liver tissue sections. Furthermore, our results indicate that 15% of the injected PEO-PGA coated maghemite nanoparticles circulate in the blood compartment for at least 60 min after i.v. application

Superparamagnetic relaxation of weakly interacting particles

DEFF Research Database (Denmark)

Mørup, Steen; Tronc, Elisabeth

1994-01-01

The influence of particle interactions on the superparamagnetic relaxation time has been studied by Mossbauer spectroscopy in samples of maghemite (gamma-Fe2O3) particles with different particle sizes and particle separations. It is found that the relaxation time decreases with decreasing particl...

NeutrAvidin Functionalization of CdSe/CdS Quantum Nanorods and Quantification of Biotin Binding Sites using Biotin-4-Fluorescein Fluorescence Quenching.

Science.gov (United States)

Lippert, Lisa G; Hallock, Jeffrey T; Dadosh, Tali; Diroll, Benjamin T; Murray, Christopher B; Goldman, Yale E

2016-03-16

We developed methods to solubilize, coat, and functionalize with NeutrAvidin elongated semiconductor nanocrystals (quantum nanorods, QRs) for use in single molecule polarized fluorescence microscopy. Three different ligands were compared with regard to efficacy for attaching NeutrAvidin using the "zero-length cross-linker" 1-ethyl-3-[3-(dimethylamino)propyl]carbodiimide (EDC). Biotin-4-fluorescene (B4F), a fluorophore that is quenched when bound to avidin proteins, was used to quantify biotin binding activity of the NeutrAvidin coated QRs and biotin binding activity of commercially available streptavidin coated quantum dots (QDs). All three coating methods produced QRs with NeutrAvidin coating density comparable to the streptavidin coating density of the commercially available quantum dots (QDs) in the B4F assay. One type of QD available from the supplier (ITK QDs) exhibited ∼5-fold higher streptavidin surface density compared to our QRs, whereas the other type of QD (PEG QDs) had 5-fold lower density. The number of streptavidins per QD increased from ∼7 streptavidin tetramers for the smallest QDs emitting fluorescence at 525 nm (QD525) to ∼20 tetramers for larger, longer wavelength QDs (QD655, QD705, and QD800). QRs coated with NeutrAvidin using mercaptoundecanoicacid (MUA) and QDs coated with streptavidin bound to biotinylated cytoplasmic dynein in single molecule TIRF microscopy assays, whereas Poly(maleic anhydride-alt-1-ocatdecene) (PMAOD) or glutathione (GSH) QRs did not bind cytoplasmic dynein. The coating methods require optimization of conditions and concentrations to balance between substantial NeutrAvidin binding vs tendency of QRs to aggregate and degrade over time.

Microfluidic magnetic bead conveyor belt

NARCIS (Netherlands)

van Pelt, S.; Frijns, A.J.H.; den Toonder, J.M.J.

2017-01-01

Magnetic beads play an important role in the miniaturization of clinical diagnostics systems. In lab-on-chip platforms, beads can be made to link to a target species and can then be used for the manipulation and detection of this species. Current bead actuation systems utilize complex on-chip coil

Flexibility contra Stiffness: The Phragmoplast as a Physical Barrier for Beads But Not for Vesicles[OA

Science.gov (United States)

Esseling-Ozdoba, Agnieszka; Kik, Richard A.; van Lammeren, André A.M.; Kleijn, J. Mieke; Emons, Anne Mie C.

2010-01-01

In plant cells, Golgi vesicles are transported to the division plane to fuse with each other, forming the cell plate, the initial membrane-bordered cell wall separating daughter cells. Vesicles, but not organelles, move through the phragmoplast, which consists of two opposing cylinders of microtubules and actin filaments, interlaced with endoplasmic reticulum membrane. To study physical aspects of this transport/inhibition process, we microinjected fluorescent synthetic 1,2-dioleoyl-sn-glycero-3-phospho-rac-1-glycerol (DOPG) vesicles and polystyrene beads into Tradescantia virginiana stamen hair cells. The phragmoplast was nonselective for DOPG vesicles of a size up to 150 nm in diameter but was a physical barrier for polystyrene beads having a diameter of 20 and 40 nm and also when beads were coated with the same DOPG membrane. We conclude that stiffness is a parameter for vesicle transit through the phragmoplast and discuss that cytoskeleton configurations can physically block such transit. PMID:19939943

Oxidative stress response in neural stem cells exposed to different superparamagnetic iron oxide nanoparticles.

Science.gov (United States)

Pongrac, Igor M; Pavičić, Ivan; Milić, Mirta; Brkić Ahmed, Lada; Babič, Michal; Horák, Daniel; Vinković Vrček, Ivana; Gajović, Srećko

2016-01-01

Biocompatibility, safety, and risk assessments of superparamagnetic iron oxide nanoparticles (SPIONs) are of the highest priority in researching their application in biomedicine. One improvement in the biological properties of SPIONs may be achieved by different functionalization and surface modifications. This study aims to investigate how a different surface functionalization of SPIONs - uncoated, coated with d-mannose, or coated with poly-l-lysine - affects biocompatibility. We sought to investigate murine neural stem cells (NSCs) as important model system for regenerative medicine. To reveal the possible mechanism of toxicity of SPIONs on NSCs, levels of reactive oxygen species, intracellular glutathione, mitochondrial membrane potential, cell-membrane potential, DNA damage, and activities of SOD and GPx were examined. Even in cases where reactive oxygen species levels were significantly lowered in NSCs exposed to SPIONs, we found depleted intracellular glutathione levels, altered activities of SOD and GPx, hyperpolarization of the mitochondrial membrane, dissipated cell-membrane potential, and increased DNA damage, irrespective of the surface coating applied for SPION stabilization. Although surface coating should prevent the toxic effects of SPIONs, our results showed that all of the tested SPION types affected the NSCs similarly, indicating that mitochondrial homeostasis is their major cellular target. Despite the claimed biomedical benefits of SPIONs, the refined determination of their effects on various cellular functions presented in this work highlights the need for further safety evaluations. This investigation helps to fill the knowledge gaps on the criteria that should be considered in evaluating the biocompatibility and safety of novel nanoparticles.

A Quantitative Electrochemiluminescence Assay for Clostridium perfringens alpha toxin

National Research Council Canada - National Science Library

Merrill, Gerald A; Rivera, Victor R; Neal, Dwayne D; Young, Charles; Poli, Mark A

2006-01-01

.... Biotinylated antibodies to C. perfringens alpha toxin bound to streptavidin paramagnetic beads specifically immunoadsorbed soluble sample alpha toxin which subsequently selectively immunoadsorbed ruthenium (Ru...

The control of beads diameter of bead-on-string electrospun nanofibers and the corresponding release behaviors of embedded drugs

Energy Technology Data Exchange (ETDEWEB)

Li, Tingxiao [Key Laboratory of Textile Science and Technology (Donghua University), Ministry of Education of China, Shanghai 201620 (China); College of Textiles, Donghua University, Shanghai 201620 (China); Ding, Xin, E-mail: xding@dhu.edu.cn [Key Laboratory of Textile Science and Technology (Donghua University), Ministry of Education of China, Shanghai 201620 (China); College of Textiles, Donghua University, Shanghai 201620 (China); Tian, Lingling, E-mail: lingling_tian@nus.edu.sg [Center of Nanofibers & Nanotechnology, Department of Mechanical Engineering, National University of Singapore, 117576 (Singapore); Hu, Jiyong; Yang, Xudong [Key Laboratory of Textile Science and Technology (Donghua University), Ministry of Education of China, Shanghai 201620 (China); College of Textiles, Donghua University, Shanghai 201620 (China); Ramakrishna, Seeram [Center of Nanofibers & Nanotechnology, Department of Mechanical Engineering, National University of Singapore, 117576 (Singapore); Guangdong-Hongkong-Macau Institute of CNS Regeneration (GHMICR), Jinan University, Guangzhou 510632 (China)

2017-05-01

Bead-on-string nanofibers, with appropriate control of the beads diameter, are potential fibrous structures for efficient encapsulation of particle drugs in micron scales and could achieve controlled drug release for tissue engineering applications. In this study, the beads diameter of electrospun bead-on-string nanofibers was controlled by adjusting the concentration of spinning polymer, poly (lactic-co-glycolic acid) (PLGA), and the solvent ratio of chloroform to acetone. The images of the scanning electron microscopy (SEM) suggested that bead-on-string nanofibers could be successfully obtained only with a certain range of PLGA solution concentration. Moreover, with the decrease in the solvent ratio of chloroform to acetone, the range was left-shifted towards a smaller concentration. In addition, increase in the PLGA solution concentration within the range the beads diameter became greater and the shape of the beads changed from oval to slender when increasing the PLGA concentration within the range. The bead-on-string nanofibers with different beads diameter were further used to load micro-particle drugs of tetracycline hydrochloride, as a model drug, to examine the release behavior of nanofibers scaffold. The release profiles of drug loaded bead-on-string nanofibers demonstrated the possibility to alleviate the burst drug release by means of beads diameter control. - Highlights: • Bead diameter of bead-on-string electrospun nanofibers was controlled by varying solvent ratio and polymer concentration. • The effect of the addition of particle drugs on BD of bead-on-string electrospun nanofibers was studied. • The corresponding release behaviors of nanofibers with different BD loading micro-particle drugs were investigated. • Bead-on-string nanofibers with bigger BD could alleviate the initial burst release.

The control of beads diameter of bead-on-string electrospun nanofibers and the corresponding release behaviors of embedded drugs

International Nuclear Information System (INIS)

Li, Tingxiao; Ding, Xin; Tian, Lingling; Hu, Jiyong; Yang, Xudong; Ramakrishna, Seeram

2017-01-01

Bead-on-string nanofibers, with appropriate control of the beads diameter, are potential fibrous structures for efficient encapsulation of particle drugs in micron scales and could achieve controlled drug release for tissue engineering applications. In this study, the beads diameter of electrospun bead-on-string nanofibers was controlled by adjusting the concentration of spinning polymer, poly (lactic-co-glycolic acid) (PLGA), and the solvent ratio of chloroform to acetone. The images of the scanning electron microscopy (SEM) suggested that bead-on-string nanofibers could be successfully obtained only with a certain range of PLGA solution concentration. Moreover, with the decrease in the solvent ratio of chloroform to acetone, the range was left-shifted towards a smaller concentration. In addition, increase in the PLGA solution concentration within the range the beads diameter became greater and the shape of the beads changed from oval to slender when increasing the PLGA concentration within the range. The bead-on-string nanofibers with different beads diameter were further used to load micro-particle drugs of tetracycline hydrochloride, as a model drug, to examine the release behavior of nanofibers scaffold. The release profiles of drug loaded bead-on-string nanofibers demonstrated the possibility to alleviate the burst drug release by means of beads diameter control. - Highlights: • Bead diameter of bead-on-string electrospun nanofibers was controlled by varying solvent ratio and polymer concentration. • The effect of the addition of particle drugs on BD of bead-on-string electrospun nanofibers was studied. • The corresponding release behaviors of nanofibers with different BD loading micro-particle drugs were investigated. • Bead-on-string nanofibers with bigger BD could alleviate the initial burst release.

Superparamagnetic relaxation in alpha-Fe particles

DEFF Research Database (Denmark)

Bødker, Franz; Mørup, Steen; Pedersen, Michael Stanley

1998-01-01

The superparamagnetic relaxation time of carbon-supported alpha-Fe particles with an average size of 3.0 Mm has been studied over a large temperature range by the use of Mossbauer spectroscopy combined with AC and DC magnetization measurements. It is found that the relaxation time varies...

Intracellular Delivery of siRNA by Polycationic Superparamagnetic Nanoparticles

Directory of Open Access Journals (Sweden)

Betzaida Castillo

2012-01-01

Full Text Available The siRNA transfection efficiency of nanoparticles (NPs, composed of a superparamagnetic iron oxide core modified with polycationic polymers (poly(hexamethylene biguanide or branched polyethyleneimine, were studied in CHO-K1 and HeLa cell lines. Both NPs demonstrated to be good siRNA transfection vehicles, but unmodified branched polyethyleneimine (25 kD was superior on both cell lines. However, application of an external magnetic field during transfection (magnetofection increased the efficiency of the superparamagnetic NPs. Furthermore, our results reveal that these NPs are less toxic towards CHO-K1 cell lines than the unmodified polycationic-branched polyethyleneimine (PEI. In general, the external magnetic field did not alter the cell’s viability nor it disrupted the cell membranes, except for the poly(hexamethylene biguanide-modified NP, where it was observed that in CHO-K1 cells application of the external magnetic field promoted membrane damage. This paper presents new polycationic superparamagnetic NPs as promising transfection vehicles for siRNA and demonstrates the advantages of magnetofection.

Fused Bead Analysis of Diogenite Meteorites

Science.gov (United States)

Mittlefehldt, D.W.; Beck, B.W.; McSween, H.Y.; Lee, C.T. A.

2009-01-01

Bulk rock chemistry is an essential dataset in meteoritics and planetary science [1]. A common method used to obtain the bulk chemistry of meteorites is ICP-MS. While the accuracy, precision and low detection limits of this process are advantageous [2], the sample size used for analysis (approx.70 mg) can be a problem in a field where small and finite samples are the norm. Fused bead analysis is another bulk rock analytical technique that has been used in meteoritics [3]. This technique involves forming a glass bead from 10 mg of sample and measuring its chemistry using a defocused beam on a microprobe. Though the ICP-MS has lower detection limits than the microprobe, the fused bead method destroys a much smaller sample of the meteorite. Fused bead analysis was initially designed for samples with near-eutectic compositions and low viscosities. Melts generated of this type homogenize at relatively low temperatures and produce primary melts near the sample s bulk composition [3]. The application of fused bead analysis to samples with noneutectic melt compositions has not been validated. The purpose of this study is to test if fused bead analysis can accurately determine the bulk rock chemistry of non-eutectic melt composition meteorites. To determine this, we conduct two examinations of the fused bead. First, we compare ICP-MS and fused bead results of the same samples using statistical analysis. Secondly, we inspect the beads for the presence of crystals and chemical heterogeneity. The presence of either of these would indicate incomplete melting and quenching of the bead.

Preparation and characterization of PVPI-coated Fe3O4 nanoparticles as an MRI contrast agent

International Nuclear Information System (INIS)

Wang, Guangshuo; Chang, Ying; Wang, Ling; Wei, Zhiyong; Kang, Jianyun; Sang, Lin; Dong, Xufeng; Chen, Guangyi; Wang, Hong; Qi, Min

2013-01-01

Polyvinylpyrrolidone-iodine (PVPI)-coated Fe 3 O 4 nanoparticles were prepared by using inverse chemical co-precipitation method, in which the PVPI serves as a stabilizer and dispersant. The wide angle X-ray diffraction (WAXD) and selected area electron diffraction (SAED) results showed that the inverse spinel structure pure phase polycrystalline Fe 3 O 4 was obtained. The scanning electron microscopy (SEM) and transmission electron microscopy (TEM) results exhibited that the resulted Fe 3 O 4 nanoparticles were roughly spherical in shape with narrow size distribution and homogenous shape. Fourier transform infrared spectroscopy (FTIR) results suggested that PVPI interacted with Fe 3 O 4 via its carbonyl groups. Results of superconducting quantum interference device (SQUID) indicated prepared Fe 3 O 4 nanoparticles exhibited superparamagnetic behavior and high saturation magnetization. T 2 -weighted MRI images of PVPI-coated Fe 3 O 4 nanoparticles showed that the magnetic resonance signal was enhanced significantly with increasing nanoparticles concentration in water at room temperature. These results indicated that the PVPI-coated Fe 3 O 4 nanoparticles had great potential for application in MRI as a T 2 contrast agent. - Highlights: • PVPI-coated Fe 3 O 4 nanoparticles were prepared using inverse co-precipitation method. • Resulted Fe 3 O 4 nanoparticles were roughly spherical in shape with narrow size distribution and homogenous shape. • Prepared Fe 3 O 4 nanoparticles exhibited superparamagnetic behavior. • T 2 -weighted MRI images of PVPI-coated Fe 3 O 4 nanoparticles were obtained

Magnetic bead detection using nano-transformers

Energy Technology Data Exchange (ETDEWEB)

Kim, Hyung Kwon; Ahn, Doyeol [Institute of Quantum Information Processing and Systems, University of Seoul, 90 Jeonnong, Dongdaemun, Seoul 130-743 (Korea, Republic of); Hwang, Jong Seung; Hwang, Sung Woo, E-mail: dahn@uos.ac.kr [Research Center for Time-domain Nano-functional Devices and School of Electrical Engineering, Korea University, 5-1 Anam, Sungbuk, Seoul 136-701 (Korea, Republic of)

2010-11-19

A novel scheme to detect magnetic beads using a nano-scale transformer with a femtoweber resolution is reported. We have performed a Faraday's induction experiment with the nano-transformer at room temperature. The transformer shows the linear output voltage responses to the sinusoidal input current. When magnetic beads are placed on the transformer, the output responses are increased by an amount corresponding to the added magnetic flux from the beads when compared with the case of no beads on the transformer. In this way, we could determine whether magnetic beads are on top of the transformer in a single particle level.

Magnetic bead detection using nano-transformers.

Science.gov (United States)

Kim, Hyung Kwon; Hwang, Jong Seung; Hwang, Sung Woo; Ahn, Doyeol

2010-11-19

A novel scheme to detect magnetic beads using a nano-scale transformer with a femtoweber resolution is reported. We have performed a Faraday's induction experiment with the nano-transformer at room temperature. The transformer shows the linear output voltage responses to the sinusoidal input current. When magnetic beads are placed on the transformer, the output responses are increased by an amount corresponding to the added magnetic flux from the beads when compared with the case of no beads on the transformer. In this way, we could determine whether magnetic beads are on top of the transformer in a single particle level.

Preparation, Characterization and Tests of Incorporation in Stem Cells of Superparamagnetic Iron Oxide

International Nuclear Information System (INIS)

Haddad, P S; Britos, T N; Li, L M; Li, L D S

2015-01-01

Superparamagnetic iron oxide nanoparticles (SPIONs) have been produced and used as contrast-enhancing agents in magnetic resonance imaging (MRI) for diagnostic use in a wide range of maladies including cardiovascular, neurological disorders, and cancer. The reasons why these SPIONs are attractive for medical purposes are based on their important and unique features. The large surface area of the nanoparticles and their manipulation through an external magnetic field are features that allow their use for carrying a large number of molecules such as biomolecules or drugs. In this scenario, the present work reports on the synthesis and characterization of SPIONs and in vitro MRI experiments to increase their capacity as probes for MRI applications on stem cells therapy. Initially, the SPIONs were prepared through the co-precipitation method using ferrous and ferric chlorides in acidic solution. The SPIONs were coated with two thiolmolecules such as mercaptosuccinic acid (MSA) and cysteine (Cys) (molar ratio SPIONs:ligand = 1:20), leading to the formation of a stable aqueous dispersion of thiolated nanoparticles (SH-SPIONs). The SH-SPIONs were characterized by Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), transmission electron microscopy (TEM), and vibrating sample magnetometry (VSM). The results showed that the SH-SPIONs have a mean diameter of 14 nm and display superparamagnetic behavior at room temperature. Preliminary tests of incorporation of SH-SPIONs were evaluated stem cells. The results showed that the thiolated nanoparticles have no toxic effects for stem cells and successfully internalized and enhance the contrast in MRI. (paper)

Development of a magnetic bead-based method for the collection of circulating extracellular vesicles.

Science.gov (United States)

Shih, Chun-Liang; Chong, Kowit-Yu; Hsu, Shih-Che; Chien, Hsin-Jung; Ma, Ching-Ting; Chang, John Wen-Cheng; Yu, Chia-Jung; Chiou, Chiuan-Chian

2016-01-25

Cells release different types of extracellular vesicles (EVs). These EVs contain biomolecules, including proteins and nucleic acids, from their parent cells, which can be useful for diagnostic applications. The aim of this study was to develop a convenient procedure to collect circulating EVs with detectable mRNA or other biomolecules. Magnetic beads coated with annexin A5 (ANX-beads), which bound to phosphatidylserine moieties on the surfaces of most EVs, were tested for their ability to capture induced apoptotic bodies in vitro and other phosphatidylserine-presenting vesicles in body fluids. Our results show that up to 60% of induced apoptotic bodies could be captured by the ANX-beads. The vesicles captured from cultured media or plasma contained amplifiable RNA. Suitable blood samples for EV collection included EDTA-plasma and serum but not heparin-plasma. In addition, EVs in plasma were labile to freeze-and-thaw cycles. In rodents xenografted with human cancer cells, tumor-derived mRNA could be detected in EVs captured from serum samples. Active proteins could be detected in EVs captured from ascites but not from plasma. In conclusion, we have developed a magnetic bead-based procedure for the collection of EVs from body fluids and proved that captured EVs contain biomolecules from their parent cells, and therefore have great potential for disease diagnosis. Copyright © 2015 Elsevier B.V. All rights reserved.

Biosensing utilizing the motion of magnetic microparticles in a microfluidic system

KAUST Repository

Giouroudi, Ioanna

2010-10-23

The study for the design of a compact and inexpensive biosensing device, which can be operated either by primary care personnel or by patients as opposed to skilled operators, is presented. The main parts of the proposed device are a microfluidic channel, permanent magnets and functionalized magnetic microparticles. The innovative aspect of the proposed biosensing method is that it utilizes the volumetric increase of magnetic microparticles when analyte binds to their surface. Their velocity decreases drastically when they are accelerated by an externally applied magnetic force within a microfluidic channel. This effect is utilized to detect the presence of analyte e.g. microbes. Analytical calculations showed that a decrease in velocity of approximately 23% can be achieved due to the volumetric change of a magnetic microparticle of View the MathML source1μm diameter when HIV virions of approximately View the MathML source0,135μm are bound to its surface and by keeping its magnetic properties the same. Preliminary experiments were carried out utilizing superparamagnetic microparticles coated with streptavidin and polystyrene microparticles coated with biotin.

Effects of Superparamagnetic Nanoparticle Clusters on the Polymerase Chain Reaction

Directory of Open Access Journals (Sweden)

Toshiaki Higashi

2012-04-01

Full Text Available The polymerase chain reaction (PCR method is widely used for the reproduction and amplification of specific DNA segments, and a novel PCR method using nanomaterials such as gold nanoparticles has recently been reported. This paper reports on the effects of superparamagnetic nanoparticles on PCR amplification without an external magnetic field, and clarifies the mechanism behind the effects of superparamagnetic particle clusters on PCR efficiency by estimating the structures of such clusters in PCR. It was found that superparamagnetic nanoparticles tend to inhibit PCR amplification depending on the structure of the magnetic nanoparticle clusters. The paper also clarifies that Taq polymerase is captured in the spaces formed among magnetic nanoparticle clusters, and that it is captured more efficiently as a result of their motion from heat treatment in PCR thermal cycles. Consequently, Taq polymerase that should be used in PCR is reduced in the PCR solution. These outcomes will be applied to novel PCR techniques using magnetic particles in an external magnetic field.

Streptavidin-functionalized capillary immune microreactor for highly efficient chemiluminescent immunoassay

Energy Technology Data Exchange (ETDEWEB)

Yang Zhanjun [State Key Laboratory of Analytical Chemistry for Life Science, Department of Chemistry, Nanjing University, Nanjing 210093 (China); College of Chemistry and Engineering, Yangzhou University, 88 South University Avenue, Yangzhou 225002 (China); Zong Chen [State Key Laboratory of Analytical Chemistry for Life Science, Department of Chemistry, Nanjing University, Nanjing 210093 (China); Ju Huangxian, E-mail: hxju@nju.edu.cn [State Key Laboratory of Analytical Chemistry for Life Science, Department of Chemistry, Nanjing University, Nanjing 210093 (China); Yan Feng, E-mail: yanfeng2007@sohu.com [Jiangsu Institute of Cancer Prevention and Cure, Nanjing 210009 (China)

2011-11-07

Highlights: {yields} A novel capillary immune microreactor was proposed for highly efficient flow-through chemiluminescent immunoassay. {yields} The microreactor was prepared by functionalizing capillary inner wall with streptavidin for capture of biotinylated antibody. {yields} The proposed immunoassay method showed wide dynamic range, good reproducibility, stability and practicality. {yields} The microreactor was low-cost and disposable, and possessed several advantages over the conventional immunoreactors. - Abstract: A streptavidin functionalized capillary immune microreactor was designed for highly efficient flow-through chemiluminescent (CL) immunoassay. The functionalized capillary could be used as both a support for highly efficient immobilization of antibody and a flow cell for flow-through immunoassay. The functionalized inner wall and the capture process were characterized using scanning electron microscopy. Compared to conventional packed tube or thin-layer cell immunoreactor, the proposed microreactor showed remarkable properties such as lower cost, simpler fabrication, better practicality and wider dynamic range for fast CL immunoassay with good reproducibility and stability. Using {alpha}-fetoprotein as model analyte, the highly efficient CL flow-through immunoassay system showed a linear range of 3 orders of magnitude from 0.5 to 200 ng mL{sup -1} and a low detection limit of 0.1 ng mL{sup -1}. The capillary immune microreactor could make up the shortcoming of conventional CL immunoreactors and provided a promising alternative for highly efficient flow-injection immunoassay.

Digital quantification of gene methylation in stool DNA by emulsion-PCR coupled with hydrogel immobilized bead-array.

Science.gov (United States)

Liu, Yunlong; Wu, Haiping; Zhou, Qiang; Song, Qinxin; Rui, Jianzhong; Zou, Bingjie; Zhou, Guohua

2017-06-15

Aberrations of gene methylation in stool DNA (sDNA) is an effective biomarker for non-invasive colorectal cancer diagnosis. However, it is challenging to accurately quantitate the gene methylation levels in sDNA due to the low abundance and degradation of sDNA. In this study, a digital quantification strategy was proposed by combining emulsion PCR (emPCR) with hydrogel immobilized bead-array. The assay includes following steps: bisulfite conversion of sDNA, pre-amplification by PCR with specific primers containing 5' universal sequences, emPCR of pre-amplicons with beaded primers to achieve single-molecular amplification and identification of hydrogel embedding beads coated with amplicons. The sensitivity and the specificity of the method are high enough to pick up 0.05% methylated targets from unmethylated DNA background. The successful detection of hypermethylated vimentin gene in clinical stool samples suggests that the proposed method should be a potential tool for non-invasive colorectal cancer screening. Copyright © 2016 Elsevier B.V. All rights reserved.

Streptavidin Modified ZnO Film Bulk Acoustic Resonator for Detection of Tumor Marker Mucin 1

Science.gov (United States)

Zheng, Dan; Guo, Peng; Xiong, Juan; Wang, Shengfu

2016-09-01

A ZnO-based film bulk acoustic resonator has been fabricated using a magnetron sputtering technology, which was employed as a biosensor for detection of mucin 1. The resonant frequency of the thin-film bulk acoustic resonator was located near at 1503.3 MHz. The average electromechanical coupling factor {K}_{eff}^2 and quality factor Q were 2.39 % and 224, respectively. Using the specific binding system of avidin-biotin, the streptavidin was self-assembled on the top gold electrode as the sensitive layer to indirectly test the MUC1 molecules. The resonant frequency of the biosensor decreases in response to the mass loading in range of 20-500 nM. The sensor modified with the streptavidin exhibits a high sensitivity of 4642.6 Hz/nM and a good selectivity.

Disposal of bead ion exchange resin wastes

International Nuclear Information System (INIS)

Gay, R.L.; Granthan, L.F.

1985-01-01

Bead ion exchange resin wastes are disposed of by a process which involves spray-drying a bead ion exchange resin waste in order to remove substantially all of the water present in such waste, including the water on the surface of the ion exchange resin beads and the water inside the ion exchange resin beads. The resulting dried ion exchange resin beads can then be solidified in a suitable solid matrix-forming material, such as a polymer, which solidifies to contain the dried ion exchange resin beads in a solid monolith suitable for disposal by burial or other conventional means

Porphyrin synthesized from cashew nut shell liquid as part of a novel superparamagnetic fluorescence nanosystem

Energy Technology Data Exchange (ETDEWEB)

Clemente, C. S.; Ribeiro, V. G. P.; Sousa, J. E. A.; Maia, F. J. N.; Barreto, A. C. H. [Universidade Federal do Ceara, Laboratorio de Produtos e Tecnologia em Processos (LPT) (Brazil); Andrade, N. F. [Universidade Federal do Ceara, Departamento de Fisica (Brazil); Denardin, J. C. [Universidad de Santiago de Chile (USACH), Departamento de Fisica (Chile); Mele, G. [Universita del Salento, Dipartimento di Ingegneria dell' Innovazione (Italy); Carbone, L. [NNL, Istituto Nanoscienze UOS Lecce (Italy); Mazzetto, S. E. [Universidade Federal do Ceara, Laboratorio de Produtos e Tecnologia em Processos (LPT) (Brazil); Fechine, P. B. A., E-mail: fechine@ufc.br [Universidade Federal do Ceara (UFC), Grupo de Quimica de Materiais Avancados (GQMAT), Departamento de Quimica Analitica e Fisico-Quimica (Brazil)

2013-06-15

Magnetic Fe{sub 3}O{sub 4} nanoparticles with average size approximately 11 nm were first oleic acid coated to interact with the meso-porphyrin derivative from CNSL. This procedure produced a novel superparamagnetic fluorescent nanosystem (SFN) linked by van der Waals interactions. This system was characterized by transmission electron microscope, infrared spectroscopy, thermogravimetric analysis, magnetic measurements, UV-Vis absorption, and fluorescence emission measurements. These results showed that SFN has good thermal stability, excellent magnetization, and nanosized dimensions ({approx}13 nm). It exhibited emission peaks at 668 and 725 nm with a maximum emission at 467 nm of excitation wavelength. The type of interaction between porphyrin and magnetic nanoparticles allowed to obtain a material with interesting optical properties which might be used as an imaging agent for contrast in cells as well as heterogeneous photocatalysis.

Synthesis of carboxyl superparamagnetic ultrasmall iron oxide (USPIO) nanoparticles by a novel flocculation-redispersion process

International Nuclear Information System (INIS)

Cheng Changming; Kou Geng; Wang Xiaoliang; Wang Shuhui; Gu Hongchen; Guo Yajun

2009-01-01

We report a novel flocculation-redispersion method to synthesize and purify the biocompatible superparamagnetic ultrasmall iron oxide (USPIO) nanoparticles coated with carboxyl dextran derivative. First, USPIO nanoparticles were synthesized and flocculated to form the large clusters through bridging effect of polyvinyl alcohol (PVA) during coprecipitation process. Then the flocculated USPIO was separated and purified from the solution conveniently through magnetic sedimentation. Finally, USPIO in the clusters were released again and well dispersed through electrostatic repelling effect of citric acid with the aid of ultrasonic. The dispersed carboxyl-functionalized USPIO was conjugated with the monoclonal antibodies. And it has been proved that the antibodies anchored on USPIO still retained their bioactivity after the conjugation. These results implied that the USPIO synthesized have good potential as active targeting molecular probe in biomedical application.

Electrochemical biotin detection based on magnetic beads and a new magnetic flow cell for screen printed electrode.

Science.gov (United States)

Biscay, Julien; González García, María Begoña; Costa García, Agustín

2015-01-01

The use of the first flow-cell for magnetic assays with an integrated magnet is reported here. The flow injection analysis system (FIA) is used for biotin determination. The reaction scheme is based on a one step competitive assay between free biotin and biotin labeled with horseradish peroxidase (B-HRP). The mixture of magnetic beads modified with streptavidin (Strep-MB), biotin and B-HRP is left 15 min under stirring and then a washing step is performed. After that, 100 μL of the mixture is injected and after 30s 100 μL of 3,3',5,5'-Tetramethylbenzidine (TMB) is injected and the FIAgram is recorded applying a potential of -0.2V. The linear range obtained is from 0.01 to 1 nM of biotin and the sensitivity is 758 nA/nM. The modification and cleaning of the electrode are performed in an easy way due to the internal magnet of the flow cell. Copyright © 2014 Elsevier B.V. All rights reserved.

Activity of an enzyme immobilized on superparamagnetic particles in a rotational magnetic field

Energy Technology Data Exchange (ETDEWEB)

Mizuki, Toru; Watanabe, Noriyuki; Nagaoka, Yutaka [Bio-Nano Electronics Research Centre, Toyo University, Saitama 350-8585 (Japan); Fukushima, Tadamasa [Shimadzu GLC Ltd., Phenomenex Support Centre, Tokyo 110-0016 (Japan); Morimoto, Hisao; Usami, Ron [Bio-Nano Electronics Research Centre, Toyo University, Saitama 350-8585 (Japan); Maekawa, Toru, E-mail: maekawa@toyonet.toyo.ac.jp [Bio-Nano Electronics Research Centre, Toyo University, Saitama 350-8585 (Japan)

2010-03-19

We immobilize {alpha}-amylase extracted from Bacillus Iicheniformis on the surfaces of superparamagnetic particles and investigate the effect of a rotational magnetic field on the enzyme's activity. We find that the activity of the enzyme molecules immobilized on superparamagnetic particles increases in the rotational magnetic field and reaches maximum at a certain frequency. We clarify the effect of the cluster structures formed by the superparamagnetic particles on the activity. Enzyme reactions are enhanced even in a tiny volume of solution using the present method, which is very important for the development of efficient micro reactors and micro total analysis systems ({mu}-TAS).

Experimental Study on Hygrothermal Deformation of External Thermal Insulation Cladding Systems with Glazed Hollow Bead

Directory of Open Access Journals (Sweden)

Houren Xiong

2016-01-01

Full Text Available This research analyzes the thermal and strain behavior of external thermal insulation cladding systems (ETICS with Glazed Hollow Beads (GHB thermal insulation mortar under hygrothermal cycles weather test in order to measure its durability under extreme weather (i.e., sunlight and rain. Thermometers and strain gauges are placed into different wall layers to gather thermal and strain data and another instrument measures the crack dimensions after every 4 cycles. The results showed that the finishing coat shrank at early stage (elastic deformation and then the finishing coat tends to expand and become damaged at later stage (plastic deformation. The deformation of insulation layer is similar to that of the finishing coat but its variation amplitude is smaller. Deformation of substrate expanded with heat and contracted with cold due to the small temperature variation. The length and width of cracks on the finishing coat grew as the experiment progressed but with a decreasing growth rate and the cracks stopped growing around 70 cycles.

Superparamagnetic iron oxide nanoparticles exert different cytotoxic effects on cells grown in monolayer cell culture versus as multicellular spheroids

Energy Technology Data Exchange (ETDEWEB)

Theumer, Anja; Gräfe, Christine; Bähring, Franziska [Department of Hematology and Oncology, Jena University Hospital, Erlanger Allee 101, 07747 Jena (Germany); Bergemann, Christian [Chemicell GmbH, Eresburgstrasse 22–23, 12103 Berlin (Germany); Hochhaus, Andreas [Department of Hematology and Oncology, Jena University Hospital, Erlanger Allee 101, 07747 Jena (Germany); Clement, Joachim H., E-mail: joachim.clement@med.uni-jena.de [Department of Hematology and Oncology, Jena University Hospital, Erlanger Allee 101, 07747 Jena (Germany)

2015-04-15

The aim of this study was to investigate the interaction of superparamagnetic iron oxide nanoparticles (SPION) with human blood–brain barrier-forming endothelial cells (HBMEC) in two-dimensional cell monolayers as well as in three-dimensional multicellular spheroids. The precise nanoparticle localisation and the influence of the NP on the cellular viability and the intracellular Akt signalling were studied in detail. Long-term effects of different polymer-coated nanoparticles (neutral fluidMAG-D, anionic fluidMAG-CMX and cationic fluidMAG-PEI) and the corresponding free polymers on cellular viability of HBMEC were investigated by real time cell analysis studies. Nanoparticles exert distinct effects on HBMEC depending on the nanoparticles' surface charge and concentration, duration of incubation and cellular context. The most severe effects were caused by PEI-coated nanoparticles. Concentrations above 25 µg/ml led to increased amounts of dead cells in monolayer culture as well as in multicellular spheroids. On the level of intracellular signalling, context-dependent differences were observed. Monolayer cultures responded on nanoparticle incubation with an increase in Akt phosphorylation whereas spheroids on the whole show a decreased Akt activity. This might be due to the differential penetration and distribution of PEI-coated nanoparticles.

Microfluidic magnetic bead conveyor belt.

Science.gov (United States)

van Pelt, Stijn; Frijns, Arjan; den Toonder, Jaap

2017-11-07

Magnetic beads play an important role in the miniaturization of clinical diagnostics systems. In lab-on-chip platforms, beads can be made to link to a target species and can then be used for the manipulation and detection of this species. Current bead actuation systems utilize complex on-chip coil systems that offer low field strengths and little versatility. We demonstrate a novel system based on an external rotating magnetic field and on-chip soft-magnetic structures to focus the field locally. These structures were designed and optimized using finite element simulations in order to create a number of local flux density maxima. These maxima, to which the magnetic beads are attracted, move over the chip surface in a continuous way together with the rotation of the external field, resulting in a mechanism similar to that of a conveyor belt. A prototype was fabricated using PDMS molding techniques mixed with iron powder for the magnetic structures. In the subsequent experiments, a quadrupole electromagnet was used to create the rotating external field. We observed that beads formed agglomerates that rolled over the chip surface, just above the magnetic structures. Field rotation frequencies between 0.1-50 Hz were tested resulting in magnetic bead speeds of over 1 mm s -1 for the highest frequency. With this, we have shown that our novel concept works, combining a simple design and simple operation with a powerful and versatile method for bead actuation. This makes it a promising method for further research and utilization in lab-on-chip systems.

Direct friction measurement in draw bead testing

DEFF Research Database (Denmark)

Olsson, David Dam; Bay, Niels; Andreasen, Jan Lasson

2005-01-01

The application of draw beads in sheet metal stamping ensures controlled drawing-in of flange parts. Lubrication conditions in draw beads are severe due to sliding under simultaneous bending. Based on the original draw bead test design by Nine [1] comprehensive studies of friction in draw beads...... have been reported in literature. A major drawback in all these studies is that friction is not directly measured, but requires repeated measurements of the drawing force with and without relative sliding between the draw beads and the sheet material. This implies two tests with a fixed draw bead tool...... and a freely rotating tool respectively, an approach, which inevitably implies large uncertainties due to scatter in the experimental conditions. In order to avoid this problem a new draw bead test is proposed by the authors measuring the friction force acting on the tool radius directly by a build...

Atomic layer deposition of superparamagnetic and ferrimagnetic magnetite thin films

International Nuclear Information System (INIS)

Zhang, Yijun; Liu, Ming; Ren, Wei; Zhang, Yuepeng; Chen, Xing; Ye, Zuo-Guang

2015-01-01

One of the key challenges in realizing superparamagnetism in magnetic thin films lies in finding a low-energy growth way to create sufficiently small grains and magnetic domains which allow the magnetization to randomly and rapidly reverse. In this work, well-defined superparamagnetic and ferrimagnetic Fe 3 O 4 thin films are successfully prepared using atomic layer deposition technique by finely controlling the growth condition and post-annealing process. As-grown Fe 3 O 4 thin films exhibit a conformal surface and poly-crystalline nature with an average grain size of 7 nm, resulting in a superparamagnetic behavior with a blocking temperature of 210 K. After post-annealing in H 2 /Ar at 400 °C, the as-grown α−Fe 2 O 3 sample is reduced to Fe 3 O 4 phase, exhibiting a ferrimagnetic ordering and distinct magnetic shape anisotropy. Atomic layer deposition of magnetite thin films with well-controlled morphology and magnetic properties provides great opportunities for integrating with other order parameters to realize magnetic nano-devices with potential applications in spintronics, electronics, and bio-applications

Beads from Inhumation Rite Burials of Gnezdovo Burial Mound

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Dobrova Olga P.

2017-12-01

Full Text Available The beads from 33 inhumation burials at Gnezdovo burial mound are examined in the article. The beads (total 367 were crafted from stretched tube (258, stretched stick (3, winding (45, press molding (2 pcs., welding (2 pcs., and mosaic beads (9 pcs.. The burial mound contains virtually no broken beads, including the settlement's most common yellow glass beads. Besides glass beads, cornelian, crystal, amber and faience beads have been registered among the burial mound material, as well as beads crafted with metal. Apart from beads, grave inventories contained a series of pendants with a bead strung on a wire ring. The considered complexes contain five pendants of this type. Besides Gnezdovo, similar pendants have been discovered in Kiev, Timerev, Pskov and Vladimir barrows. A comparison between bead sets from Gnezdovo and Kiev burial mounds allows to conclude that the general composition and occurrence frequency of beads is identical for these burials. At the same time, beads crafted with rock crystal, cornelian and metal are more frequently discovered in Kiev inhumations.

A String of Beads

Science.gov (United States)

Mead, Kelly C.

2006-01-01

In this article, the author relates how she designed a math activity she called Beads to use in conjunction with their school's 100th day celebration. Beads has provided her kindergarten class with many opportunities to practice a variety of math skills - counting, patterning, sorting, comparing, making sets, predicting, identifying numerals,…

On-Chip Magnetic Bead Manipulation and Detection Using a Magnetoresistive Sensor-Based Micro-Chip: Design Considerations and Experimental Characterization

KAUST Repository

Gooneratne, Chinthaka Pasan; Kodzius, Rimantas; Li, Fuquan; Foulds, Ian G.; Kosel, Jü rgen

2016-01-01

The remarkable advantages micro-chip platforms offer over cumbersome, time-consuming equipment currently in use for bio-analysis are well documented. In this research, a micro-chip that includes a unique magnetic actuator (MA) for the manipulation of superparamagnetic beads (SPBs), and a magnetoresistive sensor for the detection of SPBs is presented. A design methodology, which takes into account the magnetic volume of SPBs, diffusion and heat transfer phenomena, is presented with the aid of numerical analysis to optimize the parameters of the MA. The MA was employed as a magnetic flux generator and experimental analysis with commercially available COMPEL™ and Dynabeads® demonstrated the ability of the MA to precisely transport a small number of SPBs over long distances and concentrate SPBs to a sensing site for detection. Moreover, the velocities of COMPEL™ and Dynabead® SPBs were correlated to their magnetic volumes and were in good agreement with numerical model predictions. We found that 2.8 μm Dynabeads® travel faster, and can be attracted to a magnetic source from a longer distance, than 6.2 μm COMPEL™ beads at magnetic flux magnitudes of less than 10 mT. The micro-chip system could easily be integrated with electronic circuitry and microfluidic functions, paving the way for an on-chip biomolecule quantification device

On-Chip Magnetic Bead Manipulation and Detection Using a Magnetoresistive Sensor-Based Micro-Chip: Design Considerations and Experimental Characterization

KAUST Repository

Gooneratne, Chinthaka Pasan

2016-08-26

The remarkable advantages micro-chip platforms offer over cumbersome, time-consuming equipment currently in use for bio-analysis are well documented. In this research, a micro-chip that includes a unique magnetic actuator (MA) for the manipulation of superparamagnetic beads (SPBs), and a magnetoresistive sensor for the detection of SPBs is presented. A design methodology, which takes into account the magnetic volume of SPBs, diffusion and heat transfer phenomena, is presented with the aid of numerical analysis to optimize the parameters of the MA. The MA was employed as a magnetic flux generator and experimental analysis with commercially available COMPEL™ and Dynabeads® demonstrated the ability of the MA to precisely transport a small number of SPBs over long distances and concentrate SPBs to a sensing site for detection. Moreover, the velocities of COMPEL™ and Dynabead® SPBs were correlated to their magnetic volumes and were in good agreement with numerical model predictions. We found that 2.8 μm Dynabeads® travel faster, and can be attracted to a magnetic source from a longer distance, than 6.2 μm COMPEL™ beads at magnetic flux magnitudes of less than 10 mT. The micro-chip system could easily be integrated with electronic circuitry and microfluidic functions, paving the way for an on-chip biomolecule quantification device

On-Chip Magnetic Bead Manipulation and Detection Using a Magnetoresistive Sensor-Based Micro-Chip: Design Considerations and Experimental Characterization

Directory of Open Access Journals (Sweden)

Chinthaka P. Gooneratne

2016-08-01

Full Text Available The remarkable advantages micro-chip platforms offer over cumbersome, time-consuming equipment currently in use for bio-analysis are well documented. In this research, a micro-chip that includes a unique magnetic actuator (MA for the manipulation of superparamagnetic beads (SPBs, and a magnetoresistive sensor for the detection of SPBs is presented. A design methodology, which takes into account the magnetic volume of SPBs, diffusion and heat transfer phenomena, is presented with the aid of numerical analysis to optimize the parameters of the MA. The MA was employed as a magnetic flux generator and experimental analysis with commercially available COMPEL™ and Dynabeads® demonstrated the ability of the MA to precisely transport a small number of SPBs over long distances and concentrate SPBs to a sensing site for detection. Moreover, the velocities of COMPEL™ and Dynabead® SPBs were correlated to their magnetic volumes and were in good agreement with numerical model predictions. We found that 2.8 μm Dynabeads® travel faster, and can be attracted to a magnetic source from a longer distance, than 6.2 μm COMPEL™ beads at magnetic flux magnitudes of less than 10 mT. The micro-chip system could easily be integrated with electronic circuitry and microfluidic functions, paving the way for an on-chip biomolecule quantification device.

False biochemical diagnosis of hyperthyroidism in streptavidin-biotin-based immunoassays: the problem of biotin intake and related interferences.

Science.gov (United States)

Piketty, Marie-Liesse; Polak, Michel; Flechtner, Isabelle; Gonzales-Briceño, Laura; Souberbielle, Jean-Claude

2017-05-01

Immunoassays are now commonly used for hormone measurement, in high throughput analytical platforms. Immunoassays are generally robust to interference. However, endogenous analytical error may occur in some patients; this may be encountered in biotin supplementation or in the presence of anti-streptavidin antibody, in immunoassays involving streptavidin-biotin interaction. In these cases, the interference may induce both false positive and false negative results, and simulate a seemingly coherent hormonal profile. It is to be feared that this type of errors will be more frequently observed. This review underlines the importance of keeping close interactions between biologists and clinicians to be able to correlate the hormonal assay results with the clinical picture.

Microstructural Study on Oxygen Permeated Arc Beads

Directory of Open Access Journals (Sweden)

Kuan-Heng Liu

2015-01-01

Full Text Available We simulated short circuit of loaded copper wire at ambient atmosphere and successfully identified various phases of the arc bead. A cuprous oxide flake was formed on the surface of the arc bead in the rapid solidification process, and there were two microstructural constituents, namely, Cu-κ eutectic structure and solutal dendrites. Due to the arc bead formed at atmosphere during the local equilibrium solidification process, the phase of arc bead has segregated to the cuprous oxide flake, Cu-κ eutectic, and Cu phase solutal dendrites, which are the fingerprints of the arc bead permeated by oxygen.

Butyl acetate synthesis using immobilized lipase in calcium alginate beads

International Nuclear Information System (INIS)

Mohd Zulkhairi Abdul Rahim; Lee, Pat M.; Lee, Kong H.

2008-01-01

The esterification reaction of acetic acid and n-butanol using immobilized lipase encapsulated in calcium alginate beads (Lipase - CAB) and in chitosan coated calcium alginate beads (Lipase-CCAB) in n-hexane under mild reaction conditions were studied. Effects of temperature and substrate concentration (acetic acid and n-butanol) using Lipase - CAB, Lipase - CCAB and free lipase on the esterification reaction and their thermal stability towards esterification reaction were investigated. Results of temperature studies showed that the butyl acetate conversion increased with increase of temperature and reached the highest yield of about 70% around 50 degree Celsius for both immobilized systems but the yield of product catalyzed by free enzyme decreased as temperature was increased. Thermal stabilities studies showed that the Lipase-CCAB and Lipase-CAB were stable throughout the temperature range of 30-60 degree Celsius. However, free lipase became less stable at temperatures higher than 50 degree Celsius. The substrates, n-butanol and acetic acid exerted different effects on the esterification reaction and the reaction was favoured by higher acetic acid concentration than butanol. Kinetics parameters, Km and Vmax values for both substrates and the specific activities of the three enzyme system were also determined. The beads morphology was examined using SEM. Batch-wise operational stability studies for both immobilized systems demonstrated that the immobilized lipase performed better in the batch wise reactor system than the continuous bioreactor system and that the immobilized lipase remained active for at least 5 cycles of batch wise esterification reactions. (author)

Development and optimization of water treatment reactors using TiO2-modified polymer beads with a refractive index identical to that of water

Science.gov (United States)

Myoga, Arata; Iwashita, Ryutaro; Unno, Noriyuki; Satake, Shin-ichi; Taniguchi, Jun; Yuki, Kazuhisa; Seki, Yohji

2018-06-01

Various water purification reactors were constructed using beads of TiO2-coated MEXFLON, which is a fluoropolymer exhibiting a refractive index identical to that of water. The performance of these reactors was evaluated in a recirculation experiment utilizing an aqueous solution of methylene blue. Reactor pipes (length = 150 mm, internal diameter = 10 mm) were made of a fluorinated ethylene polymer with a refractive index of 1.338 and contained 206-bead clusters. A UV lamp was used to irradiate eight reactor pipes surrounding it. The above-mentioned eight bead-packed pipes were connected both in series and in parallel, and the performances of these two reactor types were compared. A pseudo-first-order rate constant of 0.70 h- 1 was obtained for the series connection, whereas the corresponding value for the parallel connection was 1.5 times smaller, confirming the effectiveness of increasing the reaction surface by employing a larger number of beads.

Development and optimization of water treatment reactors using TiO2-modified polymer beads with a refractive index identical to that of water

Science.gov (United States)

Myoga, Arata; Iwashita, Ryutaro; Unno, Noriyuki; Satake, Shin-ichi; Taniguchi, Jun; Yuki, Kazuhisa; Seki, Yohji

2018-03-01

Various water purification reactors were constructed using beads of TiO2-coated MEXFLON, which is a fluoropolymer exhibiting a refractive index identical to that of water. The performance of these reactors was evaluated in a recirculation experiment utilizing an aqueous solution of methylene blue. Reactor pipes (length = 150 mm, internal diameter = 10 mm) were made of a fluorinated ethylene polymer with a refractive index of 1.338 and contained 206-bead clusters. A UV lamp was used to irradiate eight reactor pipes surrounding it. The above-mentioned eight bead-packed pipes were connected both in series and in parallel, and the performances of these two reactor types were compared. A pseudo-first-order rate constant of 0.70 h- 1 was obtained for the series connection, whereas the corresponding value for the parallel connection was 1.5 times smaller, confirming the effectiveness of increasing the reaction surface by employing a larger number of beads.

Quantum Dot-Based Luminescent Oxygen Channeling Assay for Potential Application in Homogeneous Bioassays.

Science.gov (United States)

Zhuang, Si-Hui; Guo, Xin-Xin; Wu, Ying-Song; Chen, Zhen-Hua; Chen, Yao; Ren, Zhi-Qi; Liu, Tian-Cai

2016-01-01

The unique photoproperties of quantum dots are promising for potential application in bioassays. In the present study, quantum dots were applied to a luminescent oxygen channeling assay. The reaction system developed in this study was based on interaction of biotin with streptavidin. Carboxyl-modified polystyrene microspheres doped with quantum dots were biotinylated and used as acceptors. Photosensitizer-doped carboxyl-modified polystyrene microspheres were conjugated with streptavidin and used as donors. The results indicated that the singlet oxygen that was released from the donor beads diffused into the acceptor beads. The acceptor beads were then exited via thioxene, and were subsequently fluoresced. To avoid generating false positives, a high concentration (0.01 mg/mL) of quantum dots is required for application in homogeneous immunoassays. Compared to a conventional luminescent oxygen channeling assay, this quantum dots-based technique requires less time, and would be easier to automate and miniaturize because it requires no washing to remove excess labels.

Automated solid-phase subcloning based on beads brought into proximity by magnetic force.

Science.gov (United States)

Hudson, Elton P; Nikoshkov, Andrej; Uhlen, Mathias; Rockberg, Johan

2012-01-01

In the fields of proteomics, metabolic engineering and synthetic biology there is a need for high-throughput and reliable cloning methods to facilitate construction of expression vectors and genetic pathways. Here, we describe a new approach for solid-phase cloning in which both the vector and the gene are immobilized to separate paramagnetic beads and brought into proximity by magnetic force. Ligation events were directly evaluated using fluorescent-based microscopy and flow cytometry. The highest ligation efficiencies were obtained when gene- and vector-coated beads were brought into close contact by application of a magnet during the ligation step. An automated procedure was developed using a laboratory workstation to transfer genes into various expression vectors and more than 95% correct clones were obtained in a number of various applications. The method presented here is suitable for efficient subcloning in an automated manner to rapidly generate a large number of gene constructs in various vectors intended for high throughput applications.

In-bead screening

DEFF Research Database (Denmark)

2013-01-01

The present invention relates to screening of one-bead-one-compound (OBOC) combinatorial libraries which is useful for the discovery of compounds displaying molecular interactions with a biological or a physicochemical system, such as substrates and inhibitors of enzymes and the like. The invention...... provides a method for screening a library of compounds for their interaction with a physico- chemical or biological system and a corresponding kit for performing the method of screening a one-bead-one-compound library of compounds....

Gentamicin coated iron oxide nanoparticles as novel antibacterial agents

Science.gov (United States)

Bhattacharya, Proma; Neogi, Sudarsan

2017-09-01

Applications of different types of magnetic nanoparticles for biomedical purposes started a long time back. The concept of surface functionalization of the iron oxide nanoparticles with antibiotics is a novel technique which paves the path for further application of these nanoparticles by virtue of their property of superparamagnetism. In this paper, we have synthesized novel iron oxide nanoparticles surface functionalized with Gentamicin. The average size of the particles, concluded from the HR-TEM images, came to be around 14 nm and 10 nm for unmodified and modified nanoparticles, respectively. The magnetization curve M(H) obtained for these nanoparticles are typical of superparamagnetic nature and having almost zero values of coercivity and remanance. The release properties of the drug coated nanoparticles were studied; obtaining an S shaped profile, indicating the initial burst effect followed by gradual sustained release. In vitro investigations against various gram positive and gram negative strains viz Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa and Bacillus subtilis indicated significant antibacterial efficiency of the drug-nanoparticle conjugate. The MIC values indicated that a small amount like 0.2 mg ml-1 of drug capped particles induce about 98% bacterial death. The novelty of the work lies in the drug capping of the nanoparticles, which retains the superparamagnetic nature of the iron oxide nanoparticles and the medical properties of the drug simultaneously, which is found to extremely blood compatible.

Development of a novel device to trap heavy metal cations: application of the specific interaction between heavy metal cation and mismatch DNA base pair.

Science.gov (United States)

Torigoe, Hidetaka; Miyakawa, Yukako; Fukushi, Miyako; Ono, Akira; Kozasa, Tetsuo

2009-01-01

We have already found that Hg(II) cation specifically binds to T:T mismatch base pair in heteroduplex DNA, which increases the melting temperature of heteroduplex DNA involving T:T mismatch base pair by about 4 degrees C. We have also found that Ag(I) cation specifically binds to C:C mismatch base pair in heteroduplex DNA, which increases the melting temperature of heteroduplex DNA involving C:C mismatch base pair by about 4 degrees C. Using the specific interaction, we developed a novel device to trap each of Hg(II) and Ag(I) cation. The device is composed of 5'-biotinylated T-rich or C-rich DNA oligonucleotides, BIO-T20: 5'-Bio-T(20)-3' or BIO-C20: 5'-Bio-C(20)-3' (Bio is a biotin), immobilized on streptavidin-coated polystylene beads. When the BIO-T20-immobilized beads were added to a solution containing Hg(II) cation, and the beads trapping Hg(II) cation were collected by centrifugation, almost all of Hg(II) cation were removed from the solution. Also, when the BIO-C20-immobilized beads were added to a solution containing Ag(I) cation, and the beads trapping Ag(I) cation were collected by centrifugation, almost all of Ag(I) cation were removed from the solution. We conclude that, using the novel device developed in this study, Hg(II) and Ag(I) cation can be effectively removed from the solution.

Towards Hypoxia-responsive Drug-eluting Embolization Beads.

Science.gov (United States)

Ashrafi, Koorosh; Heaysman, Clare L; Phillips, Gary J; Lloyd, Andrew W; Lewis, Andrew L

2017-05-30

Drug release from chemoembolization microspheres stimulated by the presence of a chemically reducing environment may provide benefits for targeting drug resistant and metastatic hypoxic tumours. A water-soluble disulfide-based bifunctional cross-linker bis(acryloyl)-(l)-cystine (BALC) was synthesised, characterised and incorporated into a modified poly(vinyl) alcohol (PVA) hydrogel beads at varying concentrations using reverse suspension polymerisation. The beads were characterised to confirm the amount of cross-linker within each formulation and its effects on the bead properties. Elemental and UV/visible spectroscopic analysis confirmed the incorporation of BALC within the beads and sizing studies showed that in the presence of a reducing agent, all bead formulations increased in mean diameter. The BALC beads could be loaded with doxorubicin hydrochloride and amounts in excess of 300mg of drug per mL of hydrated beads could be achieved but required conversion of the carboxylic acid groups of the BALC to their sodium carboxylate salt forms. Elution of doxorubicin from the beads demonstrated a controlled release via ionic exchange. Some formulations exhibited an increase in size and release of drug in the presence of a reducing agent, and therefore demonstrated the ability to respond to an in vitro reducing environment. Copyright © 2017 The Authors. Published by Elsevier B.V. All rights reserved.

Self-organizing magnetic beads for biomedical applications

International Nuclear Information System (INIS)

Gusenbauer, Markus; Kovacs, Alexander; Reichel, Franz; Exl, Lukas; Bance, Simon; Özelt, Harald; Schrefl, Thomas

2012-01-01

In the field of biomedicine magnetic beads are used for drug delivery and to treat hyperthermia. Here we propose to use self-organized bead structures to isolate circulating tumor cells using lab-on-chip technologies. Typically blood flows past microposts functionalized with antibodies for circulating tumor cells. Creating these microposts with interacting magnetic beads makes it possible to tune the geometry in size, position and shape. We developed a simulation tool that combines micromagnetics and discrete particle dynamics, in order to design micropost arrays made of interacting beads. The simulation takes into account the viscous drag of the blood flow, magnetostatic interactions between the magnetic beads and gradient forces from external aligned magnets. We developed a particle–particle particle–mesh method for effective computation of the magnetic force and torque acting on the particles. - Highlights: ► We propose to use self-organized bead structures to isolate circulating tumor cells. ► Flexible ways are important to get a high probability of catching cancer cells. ► The beads make it possible to tune the geometry in size position and shape.

MICROSTRUCTURE FEATURES OF CHROME-NICKEL COATING WELDED WITH FILLER WIRE PL AN-111 WITH A 50% OVERLAP

Directory of Open Access Journals (Sweden)

A. G. Belik

2017-04-01

Full Text Available Purpose. The paper involves investigation of microstructure features of the coating welded with filler wire PL AN-111 with a 50% beads overlap. Methodology. Wear-resistant layer was formed by means of electric arc deposit welding using filler wire PL AN-111 on the plate from steel 09G2S. Deposit welding was conducted under the following parameters: welding current is of 650-750 A; arc voltage is of 30-34 V; welding speed is of 32 m/h. Microstructure was researched with application of optical microscopies “Neophot-21”, “Nikon Eclipse M200” and electron scanning microscopy JEOL JSM-6510 LV. Microhardness of structural constituentswas measuredwithtesterFM-300 (Future-Tech under loading of 10-50 g. Findings. It is shown that the overlap of the beads leads to the formation of inhomogeneous microstructure in the cross section that varies by zones from free-carbide austenite to hypereutectic microstructure with primary chromium carbides. The analysis of the microhardness of the structural constituents in various coating areas was carried out. It was found that hardness of austenite, carbide eutectic and carbides M7C3 varies in coatings in the range of 3 100-3 850 МPа, 4 100-6 800 МPа and 12 100-15 100 МPа, accordingly. Originality. Authors determined that Cr-Ni coating comprises substantially austenitic-carbide eutectic with different density and thickness of carbide fibers within eutectic colonies. Along the border “base/coating” a single-phase austenitic layer lies which turns into a layer with a hypoeutectic structure. In the heat affected zone from beads fusion austenite disintegration with the granular carbides formation was recorded. This leads to decreasing of matrix corrosion resistance due to chromium depletion. Above the zone of beads fusion, the coating has a hypereutectic structure with the presence of large primary chromium carbides. Practical value. It is shown that deposit welding with filler wire PL AN-111 with a 50

Metal-Containing Polystyrene Beads as Standards for Mass Cytometry.

Science.gov (United States)

Abdelrahman, Ahmed I; Ornatsky, Olga; Bandura, Dmitry; Baranov, Vladimir; Kinach, Robert; Dai, Sheng; Thickett, Stuart C; Tanner, Scott; Winnik, Mitchell A

2010-01-01

We examine the suitability of metal-containing polystyrene beads for the calibration of a mass cytometer instrument, a single particle analyser based on an inductively coupled plasma ion source and a time of flight mass spectrometer. These metal-containing beads are also verified for their use as internal standards for this instrument. These beads were synthesized by multiple-stage dispersion polymerization with acrylic acid as a comonomer. Acrylic acid acts as a ligand to anchor the metal ions within the interior of the beads. Mass cytometry enabled the bead-by-bead measurement of the metal-content and determination of the metal-content distribution. Beads synthesized by dispersion polymerization that involved three stages were shown to have narrower bead-to-bead variation in their lanthanide content than beads synthesized by 2-stage dispersion polymerization. The beads exhibited insignificant release of their lanthanide content to aqueous solutions of different pHs over a period of six months. When mixed with KG1a or U937 cell lines, metal-containing polymer beads were shown not to affect the mass cytometry response to the metal content of element-tagged antibodies specifically attached to these cells.

Degeneration of biogenic superparamagnetic magnetite.

Science.gov (United States)

Li, Y-L; Pfiffner, S M; Dyar, M D; Vali, H; Konhauser, K; Cole, D R; Rondinone, A J; Phelps, T J

2009-01-01

Magnetite crystals precipitated as a consequence of Fe(III) reduction by Shewanella algae BrY after 265 h incubation and 5-year anaerobic storage were investigated with transmission electron microscopy, Mössbauer spectroscopy and X-ray diffraction. The magnetite crystals were typically superparamagnetic with an approximate size of 13 nm. The lattice constants of the 265 h and 5-year crystals are 8.4164A and 8.3774A, respectively. The Mössbauer spectra indicated that the 265 h magnetite had excess Fe(II) in its crystal-chemistry (Fe(3+) (1.990)Fe(2+) (1.015)O(4)) but the 5-year magnetite was Fe(II)-deficient in stoichiometry (Fe(3+) (2.388)Fe(2+) (0.419)O(4)). Such crystal-chemical changes may be indicative of the degeneration of superparamagnetic magnetite through the aqueous oxidization of Fe(II) anaerobically, and the concomitant oxidation of the organic phases (fatty acid methyl esters) that were present during the initial formation of the magnetite. The observation of a corona structure on the aged magnetite corroborates the anaerobic oxidation of Fe(II) on the outer layers of magnetite crystals. These results suggest that there may be a possible link between the enzymatic activity of the bacteria and the stability of Fe(II)-excess magnetite, which may help explain why stable nano-magnetite grains are seldom preserved in natural environments.

Degeneration of Biogenic Superparamagnetic Magnetite

Energy Technology Data Exchange (ETDEWEB)

Li, Dr. Yi-Liang [University of Tennessee, Knoxville (UTK); Pfiffner, Susan M. [University of Tennessee, Knoxville (UTK); Dyar, Dr. M Darby [Mount Holyoke College; Vali, Dr. Hojatolah [McGill University, Montreal, Quebec; Konhauser, Dr, Kurt [University of Alberta; Cole, David R [ORNL; Rondinone, Adam Justin [ORNL; Phelps, Tommy Joe [ORNL

2009-01-01

ABSTRACT. Magnetite crystals precipitated as a consequence of Fe(III) reduction by Shewanella algae BrY after 265 hours incubation and 5-year storage were investigated with transmission electron microscopy, M ssbauer spectroscopy and X-ray diffraction. The magnetite crystals were typically superparamagnetic with an approximate size of 13 nm. The lattice constants of the 265 hour and 5-year crystals are 8.4164 and 8.3774 , respectively. The M ssbauer spectra indicated that the 265 hour magnetite had excess Fe(II) in its crystal-chemistry (Fe3+1.9901Fe2+ 1.0149O4) but the 5-year magnetite was Fe(II)-deficient in stoichiometry (Fe3+2.3875Fe2+0.4188O4). Such crystal-hemical changes may be indicative of the degeneration of superparamagnetic magnetite through the aqueous oxidization of Fe(II) anaerobically, and the concomitant oxidation of the organic phases(fatty acid methyl esters) that were present during the initial formation of the magnetite. The observation of a corona structure on the aged magnetite corroborates the oxidation of Fe(II) on the outer layers of magnetite crystals. These results suggest that there may be a possible link between the enzymatic activity of the bacteria and the stability of Fe(II)-excess magnetite, which may help explain why stable nano-magnetite grains are seldom preserved in natural environments.

Microarc oxidized TiO2 based ceramic coatings combined with cefazolin sodium/chitosan composited drug film on porous titanium for biomedical applications.

Science.gov (United States)

Wei, Daqing; Zhou, Rui; cheng, Su; Feng, Wei; Li, Baoqiang; Wang, Yaming; Jia, Dechang; Zhou, Yu; Guo, Haifeng

2013-10-01

Porous titanium was prepared by pressureless sintering of titanium beads with diameters of 100, 200, 400 and 600 μm. The results indicated that the mechanical properties of porous titanium changed significantly with different bead diameters. Plastic deformations such as necking phenomenon and dimple structure were observed on the fracture surface of porous titanium sintered by beads with diameter of 100 μm. However, it was difficult to find this phenomenon on the porous titanium with a titanium bead diameter of 600 μm. The microarc oxidized coatings were deposited on its surface to improve the bioactivity of porous titanium. Furthermore, the cefazolin sodium/chitosan composited films were fabricated on the microarc oxidized coatings for overcoming the inflammation due to implantation, showing good slow-release ability by addition of chitosan. And the release kinetic process of cefazolin sodium in composited films could be possibly fitted by a polynomial model. Copyright © 2013 Elsevier B.V. All rights reserved.

Rapid synthesis of water-dispersible superparamagnetic iron oxide nanoparticles by a microwave-assisted route for safe labeling of endothelial progenitor cells.

Science.gov (United States)

Carenza, Elisa; Barceló, Verónica; Morancho, Anna; Montaner, Joan; Rosell, Anna; Roig, Anna

2014-08-01

We synthesize highly crystalline citrate-coated iron oxide superparamagnetic nanoparticles that are stable and readily dispersible in water by an extremely fast microwave-assisted route and investigate the uptake of magnetic nanoparticles by endothelial cells. Nanoparticles form large aggregates when added to complete endothelial cell medium. The size of the aggregates was controlled by adjusting the ionic strength of the medium. The internalization of nanoparticles into endothelial cells was then investigated by transmission electron microscopy, magnetometry and chemical analysis, together with cell viability assays. Interestingly, a sevenfold more efficient uptake was found for systems with larger nanoparticle aggregates, which also showed significantly higher magnetic resonance imaging effectiveness without compromising cell viability and functionality. We are thus presenting an example of a straightforward microwave synthesis of citrate-coated iron oxide nanoparticles for safe endothelial progenitor cell labeling and good magnetic resonance cell imaging with potential application for magnetic cell guidance and in vivo cell tracking. Copyright © 2014 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

Microfabricated Passive Magnetic Bead separators

DEFF Research Database (Denmark)

Hansen, Mikkel Fougt; Lund-Olesen, Torsten; Smistrup, Kristian

2006-01-01

The use and manipulation of functionalized magnetic beads for bioanalysis in lab-on-a-chip systems is receiving growing interest. We have developed microfluidic systems with integrated magnetic structures for the capture and release of magnetic beads. The systems are fabricated in silicon by deep...

Complement activation in vitro and reactogenicity of low-molecular weight dextran-coated SPIONs in the pig CARPA model : Correlation with physicochemical features and clinical information

NARCIS (Netherlands)

Fülöp, Tamás; Nemes, Reka; Mészáros, Tamás; Urbanics, Rudolf; Kok, Robbert Jan; Jackman, Joshua A.; Storm, G; Szebeni, János

2018-01-01

The unique magnetic properties of superparamagnetic iron oxide nanoparticles (SPIONs) have led to their increasing use in drug delivery and imaging applications. Some polymer-coated SPIONs, however, share with many other nanoparticles the potential of causing hypersensitivity reactions known as

Rapid and sensitive PCR-dipstick DNA chromatography for multiplex analysis of the oral microbiota.

Science.gov (United States)

Tian, Lingyang; Sato, Takuichi; Niwa, Kousuke; Kawase, Mitsuo; Tanner, Anne C R; Takahashi, Nobuhiro

2014-01-01

A complex of species has been associated with dental caries under the ecological hypothesis. This study aimed to develop a rapid, sensitive PCR-dipstick DNA chromatography assay that could be read by eye for multiplex and semiquantitative analysis of plaque bacteria. Parallel oligonucleotides were immobilized on a dipstick strip for multiplex analysis of target DNA sequences of the caries-associated bacteria, Streptococcus mutans, Streptococcus sobrinus, Scardovia wiggsiae, Actinomyces species, and Veillonella parvula. Streptavidin-coated blue-colored latex microspheres were to generate signal. Target DNA amplicons with an oligonucleotide-tagged terminus and a biotinylated terminus were coupled with latex beads through a streptavidin-biotin interaction and then hybridized with complementary oligonucleotides on the strip. The accumulation of captured latex beads on the test and control lines produced blue bands, enabling visual detection with the naked eye. The PCR-dipstick DNA chromatography detected quantities as low as 100 pg of DNA amplicons and demonstrated 10- to 1000-fold higher sensitivity than PCR-agarose gel electrophoresis, depending on the target bacterial species. Semiquantification of bacteria was performed by obtaining a series of chromatograms using serial 10-fold dilution of PCR-amplified DNA extracted from dental plaque samples. The assay time was less than 3 h. The semiquantification procedure revealed the relative amounts of each test species in dental plaque samples, indicating that this disposable device has great potential in analysis of microbial composition in the oral cavity and intestinal tract, as well as in point-of-care diagnosis of microbiota-associated diseases.

Rapid and Sensitive PCR-Dipstick DNA Chromatography for Multiplex Analysis of the Oral Microbiota

Directory of Open Access Journals (Sweden)

Lingyang Tian

2014-01-01

Full Text Available A complex of species has been associated with dental caries under the ecological hypothesis. This study aimed to develop a rapid, sensitive PCR-dipstick DNA chromatography assay that could be read by eye for multiplex and semiquantitative analysis of plaque bacteria. Parallel oligonucleotides were immobilized on a dipstick strip for multiplex analysis of target DNA sequences of the caries-associated bacteria, Streptococcus mutans, Streptococcus sobrinus, Scardovia wiggsiae, Actinomyces species, and Veillonella parvula. Streptavidin-coated blue-colored latex microspheres were to generate signal. Target DNA amplicons with an oligonucleotide-tagged terminus and a biotinylated terminus were coupled with latex beads through a streptavidin-biotin interaction and then hybridized with complementary oligonucleotides on the strip. The accumulation of captured latex beads on the test and control lines produced blue bands, enabling visual detection with the naked eye. The PCR-dipstick DNA chromatography detected quantities as low as 100 pg of DNA amplicons and demonstrated 10- to 1000-fold higher sensitivity than PCR-agarose gel electrophoresis, depending on the target bacterial species. Semiquantification of bacteria was performed by obtaining a series of chromatograms using serial 10-fold dilution of PCR-amplified DNA extracted from dental plaque samples. The assay time was less than 3 h. The semiquantification procedure revealed the relative amounts of each test species in dental plaque samples, indicating that this disposable device has great potential in analysis of microbial composition in the oral cavity and intestinal tract, as well as in point-of-care diagnosis of microbiota-associated diseases.

Selective in vitro anticancer effect of superparamagnetic iron oxide nanoparticles loaded in hyaluronan polymeric micelles.

Science.gov (United States)

Smejkalová, Daniela; Nešporová, Kristina; Huerta-Angeles, Gloria; Syrovátka, Jakub; Jirák, Daniel; Gálisová, Andrea; Velebný, Vladimír

2014-11-10

Due to its native origin, excellent biocompatibility and biodegradability, hyaluronan (HA) represents an attractive polymer for superparamagnetic iron oxide nanoparticles (SPION) coating. Herein, we report HA polymeric micelles encapsulating oleic acid coated SPIONs, having a hydrodynamic size of about 100 nm and SPION loading capacity of 1-2 wt %. The HA-SPION polymeric micelles were found to be selectively cytotoxic toward a number of human cancer cell lines, mainly those of colon adenocarcinoma (HT-29). The selective inhibition of cell growth was even observed when the SPION loaded HA polymeric micelles were incubated with a mixture of control and cancer cells. The selective in vitro inhibition could not be connected with an enhanced CD44 uptake or radical oxygen species formation and was rather connected with a different way of SPION intracellular release. While aggregated iron particles were visualized in control cells, nonaggregated solubilized iron oxide particles were detected in cancer cells. In vivo SPION accumulation in intramuscular tumor following an intravenous micelle administration was confirmed by magnetic resonance (MR) imaging and histological analysis. Having a suitable hydrodynamic size, high magnetic relaxivity, and being cancer specific and able to accumulate in vivo in tumors, SPION-loaded HA micelles represent a promising platform for theranostic applications.

Polythiophene-coated Fe{sub 3}O{sub 4} superparamagnetic nanocomposite: Synthesis and application as a new sorbent for solid-phase extraction

Energy Technology Data Exchange (ETDEWEB)

Tahmasebi, Elham [Department of Chemistry, Tarbiat Modares University, P.O. Box 14115-175, Tehran (Iran, Islamic Republic of); Yamini, Yadollah, E-mail: yyamini@modares.ac.ir [Department of Chemistry, Tarbiat Modares University, P.O. Box 14115-175, Tehran (Iran, Islamic Republic of); Moradi, Morteza; Esrafili, Ali [Department of Chemistry, Tarbiat Modares University, P.O. Box 14115-175, Tehran (Iran, Islamic Republic of)

2013-04-03

Graphical abstract: In the present work, polythiophene-coated Fe{sub 3}O{sub 4} nanoparticles (Fe{sub 3}O{sub 4}@PTh NPs) have been successfully synthesized. The synthesized Fe{sub 3}O{sub 4}@PTh NPs were applied as an efficient sorbent for extraction and pre-concentration of several typical plasticizer compounds from environmental water samples. Highlights: ► A novel polythiophene-coated Fe{sub 3}O{sub 4} nanoparticles (Fe{sub 3}O{sub 4}@PTh NPs) was synthesized. ► The synthesized Fe{sub 3}O{sub 4}@PTh NPs were characterized by using different instruments. ► The Fe{sub 3}O{sub 4}@PTh NPs were applied as a sorbent for extraction of several plasticizers. ► After extraction, separation of NPs from solution was achieved by a magnetic field. ► The proposed procedure was applied to analysis of the analytes in real water samples. -- Abstract: In the present work, a novel type of superparamagnetic nanosorbent, polythiophene-coated Fe{sub 3}O{sub 4} nanoparticles (Fe{sub 3}O{sub 4}@PTh NPs), have been successfully synthesized. The synthesized NPs were characterized by scanning electron microscopy (SEM), Fourier transform-infrared (FT-IR) spectroscopy, and thermal gravimetric analysis (TGA). The synthesized Fe{sub 3}O{sub 4}@PTh NPs were applied as an efficient sorbent for extraction and preconcentration of several typical plasticizer compounds (di-n-butyl phthalate (DBP), di-(2-ethylhexyl) phthalate (DEHP), and dioctyl adipate (DOA)) from environmental water samples. Separation of Fe{sub 3}O{sub 4}@PTh NPs from the aqueous solution was simply achieved by applying external magnetic field. Separation and determination of the extracted plasticizers was performed by gas chromatography–flame ionization detection (GC–FID). Several variables affecting the extraction efficiency of the analytes i.e., amount of NPs sorbent, salt concentration, extraction time, and desorption conditions were investigated and optimized. The best working conditions were as follows: amount

Gd3+ doped Mn-Zn soft ferrite nanoparticles: Superparamagnetism and its correlation with other physical properties

International Nuclear Information System (INIS)

Thakur, Prashant; Sharma, Rohit; Sharma, Vineet; Barman, P.B.; Kumar, Manoj; Barman, Dipto; Katyal, S.C.; Sharma, Pankaj

2017-01-01

Highlights: • Superparamagnetic nanoparticles of Gd doped Mn-Zn spinel ferrites synthesized by co-precipitation. • XRD and FTIR studies justify the formation of cubical spinel structure. • Maximum saturation magnetization and magnetic moment at x = 0.025. • PL spectra shows blue shift for x = 0.025, 0.075 and may be attributed to quantum confinement. - Abstract: Superparamagnetic nanoparticles are very important in biomedicine due to their various applications like drug delivery, gene delivery in the body and also used for hyperthermia. In the present work, superparamagnetic nanoparticles of Mn 0.5 Zn 0.5 Gd x Fe 2-x O 4 (x = 0, 0.025, 0.050, 0.075, 0.1) ferrites have been prepared by co-precipitation method. Thorough characterizations (XRD, FTIR, FE-SEM, EDS, VSM and fluorescence spectroscopy) have proved the formation of cubical spinel superparamagnetic nanoparticles of soft ferrites. A cation distribution has been proposed for the determination of various important theoretical parameters for these samples. With the addition of Gd 3+ nanoparticles have shown the superparamagnetism at room temperature confirmed by VSM analysis. Photoluminescence (PL) spectra shows a blue shift (for x = 0.025, 0.075) which may be due to quantum confinement.

Gd3+ doped Mn-Zn soft ferrite nanoparticles: Superparamagnetism and its correlation with other physical properties

Science.gov (United States)

Thakur, Prashant; Sharma, Rohit; Sharma, Vineet; Barman, P. B.; Kumar, Manoj; Barman, Dipto; Katyal, S. C.; Sharma, Pankaj

2017-06-01

Superparamagnetic nanoparticles are very important in biomedicine due to their various applications like drug delivery, gene delivery in the body and also used for hyperthermia. In the present work, superparamagnetic nanoparticles of Mn0.5Zn0.5GdxFe2-xO4 (x = 0, 0.025, 0.050, 0.075, 0.1) ferrites have been prepared by co-precipitation method. Thorough characterizations (XRD, FTIR, FE-SEM, EDS, VSM and fluorescence spectroscopy) have proved the formation of cubical spinel superparamagnetic nanoparticles of soft ferrites. A cation distribution has been proposed for the determination of various important theoretical parameters for these samples. With the addition of Gd3+ nanoparticles have shown the superparamagnetism at room temperature confirmed by VSM analysis. Photoluminescence (PL) spectra shows a blue shift (for x = 0.025, 0.075) which may be due to quantum confinement.

The Beads of Translation: Using Beads to Translate mRNA into a Polypeptide Bracelet

Science.gov (United States)

Dunlap, Dacey; Patrick, Patricia

2012-01-01

During this activity, by making beaded bracelets that represent the steps of translation, students simulate the creation of an amino acid chain. They are given an mRNA sequence that they translate into a corresponding polypeptide chain (beads). This activity focuses on the events and sites of translation. The activity provides students with a…

K Basin sludge/resin bead separation test report

International Nuclear Information System (INIS)

Squier, D.M.

1998-01-01

The K Basin sludge is an accumulation of fuel element corrosion products, organic and inorganic ion exchange materials, canister gasket materials, iron and aluminum corrosion products, sand, dirt and minor amounts of other organic material. The sludge will be collected and treated for storage and eventual disposal. This process will remove the large solid materials by a 1/4 inch screen. The screened material will be subjected to nitric acid in a chemical treatment process. The organic ion exchange resin beads produce undesirable chemical reactions with the nitric acid. The resin beads must be removed from the bulk material and treated by another process. An effective bead separation method must extract 95% of the resin bead mass without entraining more than 5% of the other sludge component mass. The test plan I-INF-2729, ''Organic Ion Exchange Resin Separation Methods Evaluation,'' proposed the evaluation of air lift, hydro cyclone, agitated slurry and elutriation resin bead separation methods. This follows the testing strategy outlined in section 4.1 of BNF-2574, ''Testing Strategy to Support the Development of K Basins Sludge Treatment Process''. Engineering study BNF-3128, ''Separation of Organic Ion Exchange Resins from Sludge,'' Rev. 0, focused the evaluation tests on a method that removed the fine sludge particles by a sieve and then extracted the beads by means of a elutriation column. Ninety-nine percent of the resin beads are larger than 125 microns and 98.5 percent are 300 microns and larger. Particles smaller than 125 microns make up the largest portion of sludge in the K Basins. Eliminating a large part of the sludge's non-bead component will reduce the quantity that is lifted with the resin beads in the elutriation column. Resin bead particle size distribution measurements are given in Appendix A The Engineering Testing Laboratory conducted measurements of a elutriation column's ability to extract resin beads from a sieved, non-radioactive sludge

Effect of surfactant coating on magnetic properties of Fe3O4 nanoparticles: ESR study

International Nuclear Information System (INIS)

Koeseoglu, Yueksel

2006-01-01

Magnetic properties of surfactant-coated and uncoated superparamagnetic iron oxide nanoparticles, Fe 3 O 4 (SPION) were investigated by electron spin resonance (ESR) technique. For all samples, a strong and broad single ESR signal has been observed at all temperatures. A strong temperature dependence of ESR linewidth and resonance field is observed. Also, there is a strong effect of surfactant coating on magnetic properties of Fe 3 O 4 nanoparticles. While the resonance field is decreasing by coating, the linewidth of the ESR spectra is increasing. These changes in resonance field and the linewidth are attributed to the decrease in effective magnetic moment due to a non-collinear spin structure originated from the pinning of the surface spins and coated surfactant at the interface of nanoparticles. Also, the changes are due to the contribution of the volume of the diamagnetic coating mass to the sample volume

Construction and optimization of an efficient breathing-based isothermal emulsion amplification method

Energy Technology Data Exchange (ETDEWEB)

Shen, Yanting, E-mail: shenyanting798@126.com [Research Center for Learning Science, Southeast University, Sipailou Road no. 2, Nanjing, Jiangsu Province 210096 (China); State Key Laboratory of Bioelectronics, Southeast University, Sipailou Road no. 2, Nanjing, Jiangsu Province 210096 (China); Tian, Fei, E-mail: 642807827@qq.com [Research Center for Learning Science, Southeast University, Sipailou Road no. 2, Nanjing, Jiangsu Province 210096 (China); Tu, Jing, E-mail: jtu@seu.edu.cn [State Key Laboratory of Bioelectronics, Southeast University, Sipailou Road no. 2, Nanjing, Jiangsu Province 210096 (China); Li, Rui, E-mail: lirui901113@163.com [Research Center for Learning Science, Southeast University, Sipailou Road no. 2, Nanjing, Jiangsu Province 210096 (China); Chen, Zhenzhu, E-mail: zzchen_seu@163.com [Research Center for Learning Science, Southeast University, Sipailou Road no. 2, Nanjing, Jiangsu Province 210096 (China); Bai, Yunfei, E-mail: whitecf@seu.edu.cn [State Key Laboratory of Bioelectronics, Southeast University, Sipailou Road no. 2, Nanjing, Jiangsu Province 210096 (China); Ge, Qinyu, E-mail: geqinyu@seu.edu.cn [State Key Laboratory of Bioelectronics, Southeast University, Sipailou Road no. 2, Nanjing, Jiangsu Province 210096 (China); Lu, Zuhong, E-mail: zhlu@seu.edu.cn [State Key Laboratory of Bioelectronics, Southeast University, Sipailou Road no. 2, Nanjing, Jiangsu Province 210096 (China)

2017-06-22

The reaction temperature is one of the main factors that affect the stability of emulsion PCR (emPCR). Focusing on this point, we applied the “DNA breathing” mechanism in BEAMing (Bead, Emulsion, Amplification, and Magnetic) and proposed a more stable emulsion amplification method. Compared to the conventional emPCR, this method provided excellent results. Firstly, more stable emulsion system resulted in higher percentage of single-molecular amplifications (73.17%). Secondly, an ordinary temperature-controlling device was enough. Our outcome showed that the reaction temperature of this method was not strict so that the ordinary temperature-controlling device was enough for it (the heat block sets vs. the PCR instrument: 13.140 ± 0.110 vs. 13.008 ± 0.039, P = 0.120). Thirdly, the single-biotinylated emP{sub 1} coated streptavidin beads were stable enough to be used for this method (the control temperature vs. the reaction temperature: 2967.91 ± 409.045 vs. 3026.22 ± 442.129, P = 0.334), which could replace the double-biotinylated emP{sub 1} coated beads and was benefit for saving cost. In conclusion, the method presented here with stable emulsion system, simplified temperature-controlling device, and decreased investment would be a highly streamlined and inexpensive option for future single-molecular amplification based researches. - Highlights: • A breathing-based isothermal emulsion amplification (BIEA) method was developed. • BIEA showed excellent properties compared with conventional amplification method. • Terminal breathing of DNA duplex was firstly used in emulsion amplification.

Chondroitin sulfate-polyethylenimine copolymer-coated superparamagnetic iron oxide nanoparticles as an efficient magneto-gene carrier for microRNA-encoding plasmid DNA delivery

Science.gov (United States)

Lo, Yu-Lun; Chou, Han-Lin; Liao, Zi-Xian; Huang, Shih-Jer; Ke, Jyun-Han; Liu, Yu-Sheng; Chiu, Chien-Chih; Wang, Li-Fang

2015-04-01

MicroRNA-128 (miR-128) is an attractive therapeutic molecule with powerful glioblastoma regulation properties. However, miR-128 lacks biological stability and leads to poor delivery efficacy in clinical applications. In our previous study, we demonstrated two effective transgene carriers, including polyethylenimine (PEI)-decorated superparamagnetic iron oxide nanoparticles (SPIONs) as well as chemically-conjugated chondroitin sulfate-PEI copolymers (CPs). In this contribution, we report optimized conditions for coating CPs onto the surfaces of SPIONs, forming CPIOs, for magneto-gene delivery systems. The optimized weight ratio of the CPs and SPIONs is 2 : 1, which resulted in the formation of a stable particle as a good transgene carrier. The hydrodynamic diameter of the CPIOs is ~136 nm. The gel electrophoresis results demonstrate that the weight ratio of CPIO/DNA required to completely encapsulate pDNA is >=3. The in vitro tests of CPIO/DNA were done in 293 T, CRL5802, and U87-MG cells in the presence and absence of an external magnetic field. The magnetofection efficiency of CPIO/DNA was measured in the three cell lines with or without fetal bovine serum (FBS). CPIO/DNA exhibited remarkably improved gene expression in the presence of the magnetic field and 10% FBS as compared with a gold non-viral standard, PEI/DNA, and a commercial magnetofection reagent, PolyMag/DNA. In addition, CPIO/DNA showed less cytotoxicity than PEI/DNA and PolyMag/DNA against the three cell lines. The transfection efficiency of the magnetoplex improved significantly with an assisted magnetic field. In miR-128 delivery, a microRNA plate array and fluorescence in situ hybridization were used to demonstrate that CPIO/pMIRNA-128 indeed expresses more miR-128 with the assisted magnetic field than without. In a biodistribution test, CPIO/Cy5-DNA showed higher accumulation at the tumor site where an external magnet is placed nearby.MicroRNA-128 (miR-128) is an attractive therapeutic molecule

Bead Capture on Magnetic Sensors in a Microfluidic System

DEFF Research Database (Denmark)

Østerberg, Frederik Westergaard; Dalslet, Bjarke Thomas; Damsgaard, Christian Danvad

2009-01-01

The accumulation of magnetic beads by gravitational sedimentation and magnetic capture on a planar Hall-effect sensor integrated in a microfluidic channel is studied systematically as a function of the bead concentration, the fluid flow rate, and the sensor bias current. It is demonstrated...... that the sedimentation flux is proportional to the bead concentration and has a power law relation to the fluid flow rate. The mechanisms for the bead accumulation are investigated and it is found that gravitational sedimentation dominates the bead accumulation, whereas the stability of the sedimented beads against...

DWPF GLASS BEADS AND GLASS FRIT TRANSPORT DEMONSTRATION

Energy Technology Data Exchange (ETDEWEB)

Adamson, D; Bradley Pickenheim, B

2008-11-24

DWPF is considering replacing irregularly shaped glass frit with spherical glass beads in the Slurry Mix Evaporator (SME) process to decrease the yield stress of the melter feed (a non-Newtonian Bingham Plastic). Pilot-scale testing was conducted on spherical glass beads and glass frit to determine how well the glass beads would transfer when compared to the glass frit. Process Engineering Development designed and constructed the test apparatus to aid in the understanding and impacts that spherical glass beads may have on the existing DWPF Frit Transfer System. Testing was conducted to determine if the lines would plug with the glass beads and the glass frit slurry and what is required to unplug the lines. The flow loop consisted of vertical and horizontal runs of clear PVC piping, similar in geometry to the existing system. Two different batches of glass slurry were tested: a batch of 50 wt% spherical glass beads and a batch of 50 wt% glass frit in process water. No chemicals such as formic acid was used in slurry, only water and glass formers. The glass beads used for this testing were commercially available borosilicate glass of mesh size -100+200. The glass frit was Frit 418 obtained from DWPF and is nominally -45+200 mesh. The spherical glass beads did not have a negative impact on the frit transfer system. The transferring of the spherical glass beads was much easier than the glass frit. It was difficult to create a plug with glass bead slurry in the pilot transfer system. When a small plug occurred from setting overnight with the spherical glass beads, the plug was easy to displace using only the pump. In the case of creating a man made plug in a vertical line, by filling the line with spherical glass beads and allowing the slurry to settle for days, the plug was easy to remove by using flush water. The glass frit proved to be much more difficult to transfer when compared to the spherical glass beads. The glass frit impacted the transfer system to the point

DWPF GLASS BEADS AND GLASS FRIT TRANSPORT DEMONSTRATION

International Nuclear Information System (INIS)

Adamson, D.; Pickenheim, Bradley

2008-01-01

DWPF is considering replacing irregularly shaped glass frit with spherical glass beads in the Slurry Mix Evaporator (SME) process to decrease the yield stress of the melter feed (a non-Newtonian Bingham Plastic). Pilot-scale testing was conducted on spherical glass beads and glass frit to determine how well the glass beads would transfer when compared to the glass frit. Process Engineering Development designed and constructed the test apparatus to aid in the understanding and impacts that spherical glass beads may have on the existing DWPF Frit Transfer System. Testing was conducted to determine if the lines would plug with the glass beads and the glass frit slurry and what is required to unplug the lines. The flow loop consisted of vertical and horizontal runs of clear PVC piping, similar in geometry to the existing system. Two different batches of glass slurry were tested: a batch of 50 wt% spherical glass beads and a batch of 50 wt% glass frit in process water. No chemicals such as formic acid was used in slurry, only water and glass formers. The glass beads used for this testing were commercially available borosilicate glass of mesh size -100+200. The glass frit was Frit 418 obtained from DWPF and is nominally -45+200 mesh. The spherical glass beads did not have a negative impact on the frit transfer system. The transferring of the spherical glass beads was much easier than the glass frit. It was difficult to create a plug with glass bead slurry in the pilot transfer system. When a small plug occurred from setting overnight with the spherical glass beads, the plug was easy to displace using only the pump. In the case of creating a man made plug in a vertical line, by filling the line with spherical glass beads and allowing the slurry to settle for days, the plug was easy to remove by using flush water. The glass frit proved to be much more difficult to transfer when compared to the spherical glass beads. The glass frit impacted the transfer system to the point

Weld Bead Geometry of Ni-Based Alloy Deposited by PTA Process for Pipe Conduction of Shale Gas

Science.gov (United States)

Echavarria-Figueroa, C.; García-Vázquez, F.; Ruiz-Mondragón, J.; Hernández-García, H. M.; González-González, D.; Vargas, A.

The transportation of shale gas has the problem that the piping used for the extraction does not resist the erosion generated by the amount of solids causing cracks over the surface and it is necessary to extend the life of the pipelines. Plasma transferred arc (PTA) welded coatings are used to improve the surface properties of mechanical parts. Therefore, in this paper is studied the use of Ni-based filler metal as weld bead deposits on A36 steel substrates by PTA. In order to determine the suitable conditions to ensure coating quality on the substrate a design of experiments (DOE) was determined. Welding current, feed rate, and travel speed were used as input parameters and the dilution percentage as the response variable. The composition and properties of hardfacing or overlay deposited are strongly influenced by the dilution obtained. Control of dilution is important, where typically low dilution is desirable. When the dilution is low, the final deposit composition will be closer to that of the filler metal, and the wear and corrosion resistance of the hardfacing will also be maintained. To evaluate the features on the weld beads/substrate interface a microstructural characterization was performed by using scanning electron microscopy and to evaluate the mechanical properties was carried out hardness test.

Size-controlled synthesis of superparamagnetic iron oxide nanoparticles and their surface coating by gold for biomedical applications

International Nuclear Information System (INIS)

Maleki, H.; Simchi, A.; Imani, M.; Costa, B.F.O.

2012-01-01

The size mono-dispersity, saturation magnetization, and surface chemistry of magnetic nanoparticles (NPs) are recognized as critical factors for efficient biomedical applications. Here, we performed modified water-in-oil inverse nano-emulsion procedure for preparation of stable colloidal superparamagnetic iron oxide NPs (SPIONs) with high saturation magnetization. To achieve mono-dispersed SPIONs, optimization process was probed on several important factors including molar ratio of iron salts [Fe 3+ and Fe 2+ ], the concentration of ammonium hydroxide as reducing agent, and molar ratio of water to surfactant. The biocompatibility of the obtained NPs, at various concentrations, was evaluated via MTT (3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide) assay and the results showed that the NPs were non-toxic at concentrations 3 O 4 core/Au shell particles demonstrate strong plasmon resonance absorption and can be separated from solution using an external magnetic field. Experimental data from both physical and chemical determinations of the changes in particle size, surface plasmon resonance optical band, phase components, core–shell surface composition, and magnetic properties have confirmed the formation of the mono-dispersed core–shell nanostructure. - Highlights: ► Increasing the concentration of iron salts, cubic-shape SPION NPs were formed. The magnetic saturation of the SPIONs was also increased. ► The concentration of reducing agent exhibited marginal effect on the size of SPIONs but influenced the crystallinity of the NPs. A lower magnetic saturation was obtained at higher NH 4 OH concentrations. ► Mono-dispersed SPIONs can be prepared by nano-emulsion procedure at w=23, [Fe]=2.12 M, and [NH 4 OH]=30%. Under this condition, NPs with dimension of 9±3 nm and magnetic saturation of 54 emu/g are obtained. The synthesized SPIONs exhibited acceptable biocompatibility, >80% viability after 24 h incubation in L929 cells at concentrations <0

Energy efficient bead milling of microalgae: Effect of bead size on disintegration and release of proteins and carbohydrates.

Science.gov (United States)

Postma, P R; Suarez-Garcia, E; Safi, C; Yonathan, K; Olivieri, G; Barbosa, M J; Wijffels, R H; Eppink, M H M

2017-01-01

The disintegration of three industry relevant algae (Chlorella vulgaris, Neochloris oleoabundans and Tetraselmis suecica) was studied in a lab scale bead mill at different bead sizes (0.3-1mm). Cell disintegration, proteins and carbohydrates released into the water phase followed a first order kinetics. The process is selective towards proteins over carbohydrates during early stages of milling. In general, smaller beads led to higher kinetic rates, with a minimum specific energy consumption of ⩽0.47kWhkg DW -1 for 0.3mm beads. After analysis of the stress parameters (stress number and stress intensity), it appears that optimal disintegration and energy usage for all strains occurs in the 0.3-0.4mm range. During the course of bead milling, the native structure of the marker protein Rubisco was retained, confirming the mildness of the disruption process. Copyright © 2016 The Authors. Published by Elsevier Ltd.. All rights reserved.

In situ hybridization of superparamagnetic iron-biomolecule nanoparticles.

Science.gov (United States)

Moghimi, Nafiseh; Donkor, Apraku David; Mohapatra, Mamata; Thomas, Joseph Palathinkal; Su, Zhengding; Tang, Xiaowu Shirley; Leung, Kam Tong

2014-07-23

The increase in interest in the integration of organic-inorganic nanostructures in recent years has promoted the use of hybrid nanoparticles (HNPs) in medicine, energy conversion, and other applications. Conventional hybridization methods are, however, often long, complicated, and multistepped, and they involve biomolecules and discrete nanostructures as separate entities, all of which hinder the practical use of the resulting HNPs. Here, we present a novel, in situ approach to synthesizing size-specific HNPs using Fe-biomolecule complexes as the building blocks. We choose an anticancer peptide (p53p, MW 1.8 kDa) and an enzyme (GOx, MW 160 kDa) as model molecules to demonstrate the versatility of the method toward different types of molecules over a large size range. We show that electrostatic interaction for complex formation of metal hydroxide ion with the partially charged side of biomolecule in the solution is the key to hybridization of metal-biomolecule materials. Electrochemical deposition is then used to produce hybrid NPs from these complexes. These HNPs with controllable sizes ranging from 30 nm to 3.5 μm are found to exhibit superparamagnetic behavior, which is a big challenge for particles in this size regime. As an example of greatly improved properties and functionality of the new hybrid material, in vitro toxicity assessment of Fe-GOx HNPs shows no adverse effect, and the Fe-p53p HNPs are found to selectively bind to cancer cells. The superparamagnetic nature of these HNPs (superparamagnetic even above the size regime of 15-20 nm!), their biocompatibility, and the direct integration approach are fundamentally important to biomineralization and general synthesis strategy for bioinspired functional materials.

Development of high dispersed TiO2 paste for transparent screen-printable self-cleaning coatings on glass

International Nuclear Information System (INIS)

Wang Yuanhao; Lu, Lin; Yang Hongxing; Che Quande

2013-01-01

This paper reports a cheap and facile method to fabricate transparent self-cleaning coatings on glass by screen-printing high dispersed TiO 2 paste. Three kinds of ZrO 2 beads with diameter of 2, 1, and 0.1–0.2 mm were utilized to investigate their influence on the grinding and dispersion of the commercial TiO 2 powder in the ball mill. From the SEM images, surface profiler and transmittance spectrum it could be demonstrated that the smallest ZrO 2 bead with the diameter of 0.1–0.2 mm was the best candidate to disperse the TiO 2 powder into nanoscale size to make the high dispersed TiO 2 paste which was the key factor to achieve a smooth, high transparent TiO 2 coating. The surface wettability measurement showed that all the screen-printed coatings had super hydrophilic surfaces, which was independent to the surface morphology. However, the coating with the highest transparency showed the lowest photocatalytic activity which is mainly due to the light loss.

Superparamagnetic iron oxide coated on the surface of cellulose nanospheres for the rapid removal of textile dye under mild condition

Energy Technology Data Exchange (ETDEWEB)

Qin, Yunfeng [State Key Laboratory for Modification of Chemical Fibers and Polymer Materials, and College of Material Science and Engineering, Donghua University, Shanghai 201620 (China); Qin, Zongyi, E-mail: phqin@dhu.edu.cn [State Key Laboratory for Modification of Chemical Fibers and Polymer Materials, and College of Material Science and Engineering, Donghua University, Shanghai 201620 (China); Liu, Yannan; Cheng, Miao; Qian, Pengfei [State Key Laboratory for Modification of Chemical Fibers and Polymer Materials, and College of Material Science and Engineering, Donghua University, Shanghai 201620 (China); Wang, Qian, E-mail: drwangqian23@163.com [Department of Orthopaedics, Shanghai First People' s Hospital, Shanghai Jiaotong University, 100 Haining Road, Hongkou District, Shanghai 200080 (China); Zhu, Meifang [State Key Laboratory for Modification of Chemical Fibers and Polymer Materials, and College of Material Science and Engineering, Donghua University, Shanghai 201620 (China)

2015-12-01

Graphical abstract: - Highlights: • Anchoring superparamagnetic iron oxide on the surface of cellulose nanospheres as magnetically recyclable nanocatalys. • Achieving highly efficient Fenton-like reaction on the surface of composite nanospheres for rapid removal of textile dye. • Reaching nearly 98.0% degradation of Navy blue within 5 min under mild condition. - Abstract: Magnetic composite nanoparticles (MNPs) were prepared by anchoring iron oxide (Fe{sub 3}O{sub 4}) on the surface of carboxyl cellulose nanospheres through a facile chemical co-precipitation method. The as-prepared MNPs were characterized by atomic force microscopy, transmission electron microscopy, Fourier transform infrared spectroscopy, wide-angle X-ray diffraction measurement, thermal gravity analysis and vibrating sample magnetometry. These MNPs were of a generally spherical shape with a narrow size distribution, and exhibited superparamagnetic behaviors with high saturation magnetization. High efficient removal of Navy blue in aqueous solution was demonstrated at room temperature in a Fenton-like system containing the MNPs and H{sub 2}O{sub 2}, which benefited from small particle size, large surface area, high chemical activity, and good dispersibility of the MNPs. The removal efficiency of Navy blue induced by the MNPs prepared at a weight ratio of cellulose to iron of 1:2 were 90.6% at the first minute of the degradation reaction, and 98.0% for 5 min. Furthermore, these MNPs could be efficiently recycled and reused by using an external magnetic field. The approach presented in this paper promotes the use of renewable natural resources as templates for the preparation and stabilization of various inorganic nanomaterials for the purpose of catalysis, magnetic resonance imaging, biomedical and other potential applications.

Streptavidin-biotin-based directional double Nanobody sandwich ELISA for clinical rapid and sensitive detection of influenza H5N1.

Science.gov (United States)

Zhu, Min; Gong, Xue; Hu, Yonghong; Ou, Weijun; Wan, Yakun

2014-12-20

Influenza H5N1 is one subtype of the influenza A virus which can infect human bodies and lead to death. Timely diagnosis before its breakout is vital to the human health. The current clinical biochemical diagnosis for influenza virus are still flawed, and the diagnostic kits of H5N1 are mainly based on traditional monoclonal antibodies that hardly meet the requirements for clinical applications. Nanobody is a promising tool for diagnostics and treatment due to its smallest size, high specificity and stability. In this study, a novel Nanobody-based bioassay was developed for rapid, low-cost and sensitive detection of the influenza H5N1 virus. Nanobodies specific to H5N1 virus were selected from a VHH library by phage display technology. In this system, the biotinylated Nanobody was directionally captured by streptavidin coated on ELISA plate, which can specifically capture the H5N1 virus. Another Nanobody conjugated with HRP was used as a detector. A novel directional enzyme-linked immunosorbent assay for H5N1 using specific Nanobodies was established and compared to the conventional undirected ELISA assay. We have successfully constructed a high quality phage display Nanobody library and isolated two Nanobodies against H5N1 with high affinity and specificity. These two Nanobodies were further used to prepare the biosensor detection system. This streptavidin-biotin-based directional double Nanobodies sandwich ELISA for H5N1 detection showed superiority over the commonly undirectional ELISA protocol. The linear range of detection for standards in this immunoassay was approximately 50-1000 ng/mL and the detection limit was 14.1 ng/mL. The average recoveries of H5N1 virus from human serum samples were in the range from 94.58% to 114.51%, with a coefficient of variation less than 6.5%. Collectively, these results demonstrated that the proposed detection system is an alternative diagnostic tool that enables a rapid, inexpensive, sensitive and specific detection of the

Welding zinc coated steel with a CO/sub 2/ laser

International Nuclear Information System (INIS)

Akhter, R.; Steen, W.M.

1993-01-01

Welding of zinc coated steel has been studied using a high power CO/sub 2/ laser. This process is of great interest to the manufactures of car, washing machines and other components made from sheet steel and subject to corrosion. The problem associated with the welding of zinc coated steel is the low boiling point of zinc (906C) relative to the high melting point of steel (1500C). The problem is particularly important in lap welding where the zinc layer is between the lapped sheets. Under these conditions the laser 'keyhole' will generate very high vapour pressure in the zinc layer with a consequent severe risk of vapour eruption destroying the continuity of the weld bead. Several techniques are presented for the removal of zinc vapours from the interface between the two sheets. It is shown that this problem solved by suitable gap between the sheets during lap welding. Hence full penetration welds without deterioration of the weld bead can be obtained. A theory has been presented which predicted an exact gap size needed to exhaust the zinc vapour. The gap depends upon the welding speed, zinc coating thickness and thickness of the sheet. The theory predicts the weld quality satisfactorily. (author)

Plasma Antenna Based on Superparamagnetic Nanoparticles

Science.gov (United States)

Papadopoulos, K.

2017-12-01

A novel plasma antenna for space or ground based generation and injection of whistler and Alfven waves is presented. The new antenna concept is based on recently manufactured, small (10-60 nm radius), single domain, non-interacting magnetic grains with uniaxial magnetic anisotropy, known as superparamagnetic nanoparticles (SPN), dispersed in low viscosity, non-conducting media. SPNs can be described as ensembles of non-interacting magnetic moments μ with energy E=-μB when driven by a magnetic field B, similar to ordinary paramagnets, with exception that SPNs are composed by many thousands of magnetic atoms and as result have susceptibilities comparable to ferromagnets but with zero coercivity. The Langevin function accurately describes the dynamic behavior of the magnetization in the presence of low frequency AC fields since the characteristic mechanical (Brownian) and magnetic (Neel) relaxation times are shorter than 10msecs. For ground-based applications the grains are suspended in low viscosity carrier liquids, such as water or benzne and are known as ferrofluids. For space based applications, such as wave injection from CubeSats they can be dispersed as dust in vacuum containers. Agglomeration is avoided by coating the grains by coating their surface by an appropriate surfactant molecule. The ensemble of magnetic grains is driven to rotation at the desired VLF or ELF frequency by a pair of Helmholtz like coils surrounding the grain container. The near field electric field associated with rotating magnetic field then drives currents such as were observed in Rotating Magnetic Field experiments at the UCLA/LAPD chamber [Gigliotti et al., Phys. of Plasmas 16:092106; Karavaev et al., Phys. of Plasmas 17(1):012102,2010]. The magnetic moment of the AC coil is amplified by the susceptibility χ of the SPN ensemble that depending on the grain size and material can reach values of 104-105. Preliminary estimates indicate that less than 1 kg of SPN grains and power of

In vitro and in vivo lung deposition of coated magnetic aerosol particles.

Science.gov (United States)

Xie, Yuanyuan; Longest, P Worth; Xu, Yun Hao; Wang, Jian Ping; Wiedmann, Timothy Scott

2010-11-01

The magnetic induced deposition of polydispersed aerosols composed of agglomerated superparamagnetic particles was measured with an in vitro model system and in the mouse trachea and deep lung for the purpose of investigating the potential of site specific respiratory drug delivery. Oleic acid coated superparamagnetic particles were prepared and characterized by TEM, induced magnetic moment, and iron content. The particles were dispersed in cyclohexane, aerosolized with an ultrasonic atomizer and dried by sequential reflux and charcoal columns. The fraction of iron deposited on glass tubes increased with particle size and decreasing flow rate. High deposition occurred with a small diameter tube, but the deposition fraction was largely independent of tube size at larger diameters. Results from computational fluid dynamics qualitatively agreed with the experimental results. Enhanced deposition was observed in the mouse lung but not in the trachea consistent with the analysis of the aerodynamic time allowed for deposition and required magnetic deposition time. © 2010 Wiley-Liss, Inc. and the American Pharmacists Association

Arsenic Removal from Aqueous Solution Using Pure and Metal-Doped Titania Nanoparticles Coated on Glass Beads: Adsorption and Column Studies

Directory of Open Access Journals (Sweden)

M. Ihsan Danish

2013-01-01

Full Text Available Nanosized metal oxide, Titania, provides high surface area and specific affinity for the adsorption of heavy metals, including arsenic (As, which is posing a great threat to the world population due to its carcinogenic nature. In this study, As(III adsorption was studied on pure and metal- (Ag- and Fe- doped Titania nanoparticles. The nanoparticles were synthesized by liquid impregnation method with some modifications, with crystallite size in the range of 30 to 40 nm. Band gap analysis, using Kubelka-Munk function showed a shift of absorption band from UV to visible region for the metal-doped Titania. Effect of operational parameters like dose of nanoparticles, initial As(III concentration, and pH was evaluated at 25°C. The data obtained gave a good fit with Langmuir and Freundlich isotherms and the adsorption was found to conform to pseudo-second-order kinetics. In batch studies, over 90% of arsenic removal was observed for both types of metal-doped Titania nanoparticles from a solution containing up to 2 ppm of the heavy metal. Fixed bed columns of nanoparticles, coated on glass beads, were used for As(III removal under different operating conditions. Thomas and Yoon-Nelson models were applied to predict the breakthrough curves and to find the characteristic column parameters useful for process design. The columns were regenerated using 10% NaOH solution.

Calcium Pectinate Beads Formation: Shape and Size Analysis

Directory of Open Access Journals (Sweden)

Boon-Beng Lee

2014-04-01

Full Text Available The aim of this study was to investigate the inter-relationship between process variables and the size and shape of pectin solution droplets upon detachment from a dripping tip as well as Ca-pectinate beads formed after gelation via image analysis. The sphericity factor (SF of the droplets was generally smaller than 0.05. There was no specific trend between the SF of the droplets and the pectin concentration or the dripping tip radius. The SF the beads formed from high-concentration pectin solutions and a small dripping tip was smaller than 0.05. The results show that the Reynolds number and Ohnesorge number of the droplets fall within the operating region for forming spherical beads in the shape diagram, with the exception to the lower boundary. The lower boundary of the operating region has to be revised to Oh = 2.3. This is because the critical viscosity for Ca-pectinate bead formation is higher than that of Ca-alginate beads. On the other hand, the radius of the droplets and beads increased as the dripping tip radius increased. The bead radius can easily be predicted by Tate’s law equation.

HPMA and HEMA copolymer bead interactions with eukaryotic cells

Directory of Open Access Journals (Sweden)

Cristina D. Vianna-Soares

2004-09-01

Full Text Available Two different hydrophilic acrylate beads were prepared via aqueous suspension polymerization. Beads produced of a hydroxypropyl methacrylate (HPMA and ethyleneglycol methacrylate (EDMA copolymer were obtained using a polyvinyl alcohol suspending medium. Copolymers of 2hydroxyethyl methacrylate (HEMA, methyl methacrylate (MMA and ethyleneglycol methacrylate (EDMA beads were obtained using magnesium hydroxide as the suspending agent. Following characterization by scanning electron microscopy (SEM, nitrogen sorption analysis (NSA and mercury intrusion porosimetry (MIP, the beads were cultured with monkey fibroblasts (COS7 to evaluate their ability to support cell growth, attachment and adhesion. Cell growth behavior onto small HPMA/EDMA copolymer beads and large HEMA/MMA/EDMA copolymer beads is evaluated regarding their hidrophilicity/hidrophobicity and surface roughness.

Surface streamer propagations on an alumina bead: experimental observation and numerical modeling

Science.gov (United States)

Kang, Woo Seok; Kim, Hyun-Ha; Teramoto, Yoshiyuki; Ogata, Atsushi; Lee, Jin Young; Kim, Dae-Woong; Hur, Min; Song, Young-Hoon

2018-01-01

A surface streamer in a simplified packed-bed reactor has been studied both experimentally (through time-resolved ICCD imaging) and theoretically (through two-dimensional numerical modeling). The propagation of streamers on an alumina spherical bead without catalytic coating shows three distinct phases—the generation and propagation of a primary streamer (PS) with a moderate velocity and electric field, fast PS acceleration with an enhanced electric field, and slow secondary streamer (SS) propagation. The velocity of the streamer is less than that of propagation in a gaseous media. The electric field and velocity at the streamer front are maximized when a PS propagates during the interval from the midpoint of the bead to the bottom electrode. The SS exhibits a much lower velocity and electric field compared with the PS. The PS velocity is affected by an external applied voltage, especially when it approaches the ground electrode. However, that of the SS remains constant regardless of the voltage change. The simulation shows that the PS exhibits a high electric field mainly created by the space charge induced by electrons, whereas the SS relies on ion movement with electron decay in a charge-filled thin streamer body.

In vitro transcription of a torsionally constrained template

DEFF Research Database (Denmark)

Bentin, Thomas; Nielsen, Peter E

2002-01-01

RNA polymerase (RNAP) and the DNA template must rotate relative to each other during transcription elongation. In the cell, however, the components of the transcription apparatus may be subject to rotary constraints. For instance, the DNA is divided into topological domains that are delineated...... of torsionally constrained DNA by free RNAP. We asked whether or not a newly synthesized RNA chain would limit transcription elongation. For this purpose we developed a method to immobilize covalently closed circular DNA to streptavidin-coated beads via a peptide nucleic acid (PNA)-biotin conjugate in principle...... constrained. We conclude that transcription of a natural bacterial gene may proceed with high efficiency despite the fact that newly synthesized RNA is entangled around the template in the narrow confines of torsionally constrained supercoiled DNA....

Development of a tetrameric streptavidin mutein with reversible biotin binding capability: engineering a mobile loop as an exit door for biotin.

Directory of Open Access Journals (Sweden)

Valerie J O'Sullivan

Full Text Available A novel form of tetrameric streptavidin has been engineered to have reversible biotin binding capability. In wild-type streptavidin, loop(3-4 functions as a lid for the entry and exit of biotin. When biotin is bound, interactions between biotin and key residues in loop(3-4 keep this lid in the closed state. In the engineered mutein, a second biotin exit door is created by changing the amino acid sequence of loop(7-8. This door is mobile even in the presence of the bound biotin and can facilitate the release of biotin from the mutein. Since loop(7-8 is involved in subunit interactions, alteration of this loop in the engineered mutein results in an 11° rotation between the two dimers in reference to wild-type streptavidin. The tetrameric state of the engineered mutein is stabilized by a H127C mutation, which leads to the formation of inter-subunit disulfide bonds. The biotin binding kinetic parameters (k(off of 4.28×10(-4 s(-1 and K(d of 1.9×10(-8 M make this engineered mutein a superb affinity agent for the purification of biotinylated biomolecules. Affinity matrices can be regenerated using gentle procedures, and regenerated matrices can be reused at least ten times without any observable reduction in binding capacity. With the combination of both the engineered mutein and wild-type streptavidin, biotinylated biomolecules can easily be affinity purified to high purity and immobilized to desirable platforms without any leakage concerns. Other potential biotechnological applications, such as development of an automated high-throughput protein purification system, are feasible.

Development of a Tetrameric Streptavidin Mutein with Reversible Biotin Binding Capability: Engineering a Mobile Loop as an Exit Door for Biotin

Science.gov (United States)

O'Sullivan, Valerie J.; Barrette-Ng, Isabelle; Hommema, Eric; Hermanson, Greg T.; Schofield, Mark; Wu, Sau-Ching; Honetschlaeger, Claudia; Ng, Kenneth K.-S.; Wong, Sui-Lam

2012-01-01

A novel form of tetrameric streptavidin has been engineered to have reversible biotin binding capability. In wild-type streptavidin, loop3–4 functions as a lid for the entry and exit of biotin. When biotin is bound, interactions between biotin and key residues in loop3–4 keep this lid in the closed state. In the engineered mutein, a second biotin exit door is created by changing the amino acid sequence of loop7–8. This door is mobile even in the presence of the bound biotin and can facilitate the release of biotin from the mutein. Since loop7–8 is involved in subunit interactions, alteration of this loop in the engineered mutein results in an 11° rotation between the two dimers in reference to wild-type streptavidin. The tetrameric state of the engineered mutein is stabilized by a H127C mutation, which leads to the formation of inter-subunit disulfide bonds. The biotin binding kinetic parameters (koff of 4.28×10−4 s−1 and Kd of 1.9×10−8 M) make this engineered mutein a superb affinity agent for the purification of biotinylated biomolecules. Affinity matrices can be regenerated using gentle procedures, and regenerated matrices can be reused at least ten times without any observable reduction in binding capacity. With the combination of both the engineered mutein and wild-type streptavidin, biotinylated biomolecules can easily be affinity purified to high purity and immobilized to desirable platforms without any leakage concerns. Other potential biotechnological applications, such as development of an automated high-throughput protein purification system, are feasible. PMID:22536357

Bead magnetorelaxometry with an on-chip magnetoresistive sensor

DEFF Research Database (Denmark)

Dalslet, Bjarke Thomas; Damsgaard, Christian Danvad; Donolato, Marco

2011-01-01

Magnetorelaxometry measurements on suspensions of magnetic beads are demonstrated using a planar Hall effect sensor chip embedded in a microfluidic system. The alternating magnetic field used for magnetizing the beads is provided by the sensor bias current and the complex magnetic susceptibility...... spectra are recorded as the 2nd harmonic of the sensor response. The complex magnetic susceptibility signal appears when a magnetic bead suspension is injected, it scales with the bead concentration, and it follows the Cole-Cole expression for Brownian relaxation. The complex magnetic susceptibility...... signal resembles that from conventional magnetorelaxometry done on the same samples apart from an offset in Brownian relaxation frequency. The time dependence of the signal can be rationalized as originating from sedimented beads....

Entrapment of laurel lipase in chitosan hydrogel beads.

Science.gov (United States)

Yagar, Hulya; Balkan, Ugur

2017-08-01

Laurel seed lipase was entrapped within chitosan beads with ionotropic gelatin method using tripolyphosphate (TPP) as multivalent covalent counter ion. Immobilization yield was 78%. First, optimum immobilization conditions were determined, and morphology of chitosan beads was characterized by scanning electron microscopy. Optimum pH and temperature were evaluated as 6.0 and 40 °C, respectively. The immobilized beads saved about 55% of its activities at 60° while saved about 32% at 70 °C for 30 min. V max /K m values were determined as 31.75 and 2.87 using olive oil as substrate for immobilized beads and free enzyme, respectively. Immobilized beads showed the activities during 30 days at +4 °C.

Bead-bead interaction parameters in dissipative particle dynamics: Relation to bead-size, solubility parameter, and surface tension

Science.gov (United States)

Maiti, Amitesh; McGrother, Simon

2004-01-01

Dissipative particle dynamics (DPD) is a mesoscale modeling method for simulating equilibrium and dynamical properties of polymers in solution. The basic idea has been around for several decades in the form of bead-spring models. A few years ago, Groot and Warren [J. Chem. Phys. 107, 4423 (1997)] established an important link between DPD and the Flory-Huggins χ-parameter theory for polymer solutions. We revisit the Groot-Warren theory and investigate the DPD interaction parameters as a function of bead size. In particular, we show a consistent scheme of computing the interfacial tension in a segregated binary mixture. Results for three systems chosen for illustration are in excellent agreement with experimental results. This opens the door for determining DPD interactions using interfacial tension as a fitting parameter.

Intrinsically superparamagnetic Fe-hydroxyapatite nanoparticles positively influence osteoblast-like cell behaviour

Science.gov (United States)

2012-01-01

Background Superparamagnetic nanoparticles (MNPs) have been progressively explored for their potential in biomedical applications and in particular as a contrast agent for diagnostic imaging, for magnetic drug delivery and more recently for tissue engineering applications. Considering the importance of having safe MNPs for such applications, and the essential role of iron in bone remodelling, this study developed and analysed novel biocompatible and bioreabsorbable superparamagnetic nanoparticles, that avoid the use of poorly tolerated magnetite based nanoparticles, for bone tissue engineering applications. Results MNPs were obtained by doping hydroxyapatite (HA) with Fe ions, by directly substituting Fe2+ and Fe3+ into the HA structure yielding superparamagnetic bioactive phase. In the current study, we have investigated the effects of increasing concentrations (2000 μg/ml; 1000 μg/ml; 500 μg/ml; 200 μg/ml) of FeHA MNPs in vitro using Saos-2 human osteoblast-like cells cultured for 1, 3 and 7 days with and without the exposure to a static magnetic field of 320 mT. Results demonstrated not only a comparable osteoblast viability and morphology, but increased in cell proliferation, when compared to a commercially available Ha nanoparticles, even with the highest dose used. Furthermore, FeHA MNPs exposure to the static magnetic field resulted in a significant increase in cell proliferation throughout the experimental period, and higher osteoblast activity. In vivo preliminary results demonstrated good biocompatibility of FeHA superparamagnetic material four weeks after implantation into a critical size lesion of the rabbit condyle. Conclusions The results of the current study suggest that these novel FeHA MNPs may be particularly relevant for strategies of bone tissue regeneration and open new perspectives for the application of a static magnetic field in a clinical setting of bone replacement, either for diagnostic imaging or magnetic drug delivery

Control of the interparticle spacing in superparamagnetic iron oxide nanoparticle clusters by surface ligand engineering

Science.gov (United States)

Dan, Wang; Bingbing, Lin; Taipeng, Shen; Jun, Wu; Fuhua, Hao; Chunchao, Xia; Qiyong, Gong; Huiru, Tang; Bin, Song; Hua, Ai

2016-07-01

Polymer-mediated self-assembly of superparamagnetic iron oxide (SPIO) nanoparticles allows modulation of the structure of SPIO nanocrystal cluster and their magnetic properties. In this study, dopamine-functionalized polyesters (DA-polyester) were used to directly control the magnetic nanoparticle spacing and its effect on magnetic resonance relaxation properties of these clusters was investigated. Monodisperse SPIO nanocrystals with different surface coating materials (poly(ɛ-caprolactone), poly(lactic acid)) of different molecular weights containing dopamine (DA) structure (DA-PCL2k, DA-PCL1k, DA-PLA1k)) were prepared via ligand exchange reaction, and these nanocrystals were encapsulated inside amphiphilic polymer micelles to modulate the SPIO nanocrystal interparticle spacing. Small-angle x-ray scattering (SAXS) was applied to quantify the interparticle spacing of SPIO clusters. The results demonstrated that the tailored magnetic nanoparticle clusters featured controllable interparticle spacing providing directly by the different surface coating of SPIO nanocrystals. Systematic modulation of SPIO nanocrystal interparticle spacing can regulate the saturation magnetization (M s) and T 2 relaxation of the aggregation, and lead to increased magnetic resonance (MR) relaxation properties with decreased interparticle spacing. Project supported by the National Key Basic Research Program of China (Grant No. 2013CB933903), the National Key Technology R&D Program of China (Grant No. 2012BAI23B08), and the National Natural Science Foundation of China (Grant Nos. 20974065, 51173117, and 50830107).

Three-bead steering microswimmers

Science.gov (United States)

Rizvi, Mohd Suhail; Farutin, Alexander; Misbah, Chaouqi

2018-02-01

The self-propelled microswimmers have recently attracted considerable attention as model systems for biological cell migration as well as artificial micromachines. A simple and well-studied microswimmer model consists of three identical spherical beads joined by two springs in a linear fashion with active oscillatory forces being applied on the beads to generate self-propulsion. We have extended this linear microswimmer configuration to a triangular geometry where the three beads are connected by three identical springs in an equilateral triangular manner. The active forces acting on each spring can lead to autonomous steering motion; i.e., allowing the swimmer to move along arbitrary paths. We explore the microswimmer dynamics analytically and pinpoint its rich character depending on the nature of the active forces. The microswimmers can translate along a straight trajectory, rotate at a fixed location, as well as perform a simultaneous translation and rotation resulting in complex curved trajectories. The sinusoidal active forces on the three springs of the microswimmer contain naturally four operating parameters which are more than required for the steering motion. We identify the minimal operating parameters which are essential for the motion of the microswimmer along any given arbitrary trajectory. Therefore, along with providing insights into the mechanics of the complex motion of the natural and artificial microswimmers, the triangular three-bead microswimmer can be utilized as a model for targeted drug delivery systems and autonomous underwater vehicles where intricate trajectories are involved.

Safeguards Analytical Laboratory evaluation program. Part 1. Resin bead mass spectrometry. Part 2. Results of a resin bead field experiment-Tastex-J

International Nuclear Information System (INIS)

Walker, R.L.; Smith, D.H.; Carter, J.A.; Musick, W.R.; Donohue, D.L.; Deron, S.; Asakura, Y.; Kagami, K.; Irinouchi, S.; Masui, J.

1981-01-01

The first part of this report covers background of resin bead spectrometry and the new batch resin bead method. In the original technique, about ten anion resin beads in the nitrate form were exposed to the diluted sample solution. The solution was adjusted to be a 8 M HNO 3 and to have about 1 μg U per bead. Up to 48 hours of static contact between beads and solution was required for adsorption of 1 to 3 ng Pu and U per bead to be achieved. Under these conditions, contamination was a problem at reprocessing facilities. The new batch techniques reduces the risk of contamination by handling one hundred times more U in the final diluted sample which is exposed to a proportionately larger number of beads. Moreover, it only requires ten minutes adsorption time to provide about 1000 purified samples for mass spectrometry. The amounts of Pu and U adsorbed versus time were determined and results are tabulated. The second part of this report briefly summarizes results of resin bead field tests completed at the Power Reactor and Nuclear Fuel Development Corporation (PNC) reprocessing plant in Tokai-mura, Japan. Both methods, the original small-sample resin bead and the batch technique, were investigated on spent fuel solutions. Beads were prepared at PNC and distributed to IAEA and ORNL along with dried residues for conventional mass spectrometric analysis at IAEA. Parallel measurements were made at PNC using their normal measuring routines. The U and Pu measurements of all resin and those of PNC are in excellent agreement for the batch method. Discrepancies were noted in the U measurements by the original method

Superparamagnetic and ferrimagnetic behavior of nanocrystalline ZnO(MnO)

Science.gov (United States)

Kuryliszyn-Kudelska, I.; Dobrowolski, W.; Arciszewska, M.; Romčević, N.; Romčević, M.; Hadžić, B.; Sibera, D.; Narkiewicz, U.

2018-04-01

We have studied the magnetic properties of nanocrystals of ZnO:MnO prepared by traditional wet chemistry method. The detailed structural and morphological characterization was performed. The results of systematic measurements of AC magnetic susceptibility as a function of temperature and frequency as well as DC magnetization are reported. We observed two different types of magnetic behavior depending on the concentration doping. For samples with low nominal content (up to 30 wt% of MnO), superparamagnetic behavior was observed. We attribute the observed superparamagnetism to the presence of nanosized ZnMnO3 phase. For nanocrystals doped above nominal 60 wt% of MnO ferrimagnetism was detected with TC at around 42 K. This magnetic behavior we assign to the presence of nanosized Mn3O4 phase.

Gd{sup 3+} doped Mn-Zn soft ferrite nanoparticles: Superparamagnetism and its correlation with other physical properties

Energy Technology Data Exchange (ETDEWEB)

Thakur, Prashant; Sharma, Rohit; Sharma, Vineet; Barman, P.B. [Department of Physics & Materials Science, Jaypee University of Information Technology, Waknaghat, Solan, Himachal Pradesh 173234 (India); Kumar, Manoj [Department of Physics & Materials Science, Jaypee Institute of Information Technology, Noida 201307 (India); Barman, Dipto [Gwangju Institute of Science & Technology, Gwangju (Korea, Republic of); Department of Computer Science & Engineering, Jaypee University of Information Technology, Waknaghat, Solan, Himachap Pradesh 173234 (India); Katyal, S.C. [Department of Physics & Materials Science, Jaypee Institute of Information Technology, Noida 201307 (India); Sharma, Pankaj, E-mail: pankaj.sharma@juit.ac.in [Department of Physics & Materials Science, Jaypee University of Information Technology, Waknaghat, Solan, Himachal Pradesh 173234 (India)

2017-06-15

Highlights: • Superparamagnetic nanoparticles of Gd doped Mn-Zn spinel ferrites synthesized by co-precipitation. • XRD and FTIR studies justify the formation of cubical spinel structure. • Maximum saturation magnetization and magnetic moment at x = 0.025. • PL spectra shows blue shift for x = 0.025, 0.075 and may be attributed to quantum confinement. - Abstract: Superparamagnetic nanoparticles are very important in biomedicine due to their various applications like drug delivery, gene delivery in the body and also used for hyperthermia. In the present work, superparamagnetic nanoparticles of Mn{sub 0.5}Zn{sub 0.5}Gd{sub x}Fe{sub 2-x}O{sub 4} (x = 0, 0.025, 0.050, 0.075, 0.1) ferrites have been prepared by co-precipitation method. Thorough characterizations (XRD, FTIR, FE-SEM, EDS, VSM and fluorescence spectroscopy) have proved the formation of cubical spinel superparamagnetic nanoparticles of soft ferrites. A cation distribution has been proposed for the determination of various important theoretical parameters for these samples. With the addition of Gd{sup 3+} nanoparticles have shown the superparamagnetism at room temperature confirmed by VSM analysis. Photoluminescence (PL) spectra shows a blue shift (for x = 0.025, 0.075) which may be due to quantum confinement.

EGF receptor targeted tumor imaging with biotin-PEG-EGF linked to 99mTc-HYNIC labeled avidin and streptavidin

International Nuclear Information System (INIS)

Jung, Kyung-Ho; Park, Jin Won; Paik, Jin-Young; Quach, Cung Hoa Thien; Choe, Yearn Seong; Lee, Kyung-Han

2012-01-01

Introduction: As direct radiolabeled peptides suffer limitations for in vivo imaging, we investigated the usefulness of radioloabeled avidin and streptavidin as cores to link peptide ligands for targeted tumor imaging. Methods: Human epidermal growth factor (EGF) was site specifically conjugated with a single PEG-biotin molecule and linked to 99m Tc-HYNIC labeled avidin-FITC (Av) or streptavidin-Cy5.5 (Sav). Receptor targeting was verified in vitro, and in vivo pharmacokinetic and biodistribution profiles were studied in normal mice. Scintigraphic imaging was performed in MDA-MB-468 breast tumor xenografted nude mice. Results: Whereas both 99m Tc-Av-EGF and 99m Tc-Sav-EGF retained receptor-specific binding in vitro, the two probes substantially diverged in pharmacokinetic and biodistribution behavior in vivo. 99m Tc-Av-EGF was rapidly eliminated from the circulation with a T1/2 of 4.3 min, and showed intense hepatic accumulation but poor tumor uptake (0.6%ID/gm at 4 h). 99m Tc-Sav-EGF displayed favorable in vivo profiles of longer circulation (T1/2β, 51.5 min) and lower nonspecific uptake that resulted in higher tumor uptake (3.8 %ID/gm) and clear tumor visualization at 15 h. Conclusion: 99m Tc-HYNIC labeled streptavidin linked with growth factor peptides may be useful as a protein-ligand complex for targeted imaging of tumor receptors.

Tunable bead-on-string microstructures fabricated by mechano-electrospinning

International Nuclear Information System (INIS)

Bu Ningbin; Huang Yongan; Deng Huixu; Yin Zhouping

2012-01-01

In this paper, bead-on-string microstructures are fabricated by the mechano-electrospinning (MES) process in a continuously tunable manner. The thin jet is pulled onto the substrate by the stable electric field force and tunable mechanical drawing force, and then the bead-on-string structures are generated by means of the force exerted on the jet, which changes from capillary force and resisting viscosity force to friction force at the contact point in the horizontal direction. In a stable bead-on-string formation process, one cycle can be divided into three stages from the point of view of the jet behaviour: being anchored, being stretched, and skipping. The bead size and the bead gap are continuously tunable through the MES process. The fabrication mechanisms of the bead-on-string microstructure are uncovered through theoretical analysis and experimental characterization. When a critical velocity is achieved, the jet directly falls on the substrate without accumulation since the mechanical drawing force in the horizontal direction overtakes the capillary force, which leads the bead-on-string microstructures to a continuous fibre line. It is a flexible and highly controllable method to fabricate bead-on-string microstructures.

A Controlled Drug-Delivery Experiment Using Alginate Beads

Science.gov (United States)

Farrell, Stephanie; Vernengo, Jennifer

2012-01-01

This paper describes a simple, cost-effective experiment which introduces students to drug delivery and modeling using alginate beads. Students produce calcium alginate beads loaded with drug and measure the rate of release from the beads for systems having different stir rates, geometries, extents of cross-linking, and drug molecular weight.…

One-Step Method for Preparation of Magnetic Nanoparticles Coated with Chitosan

Directory of Open Access Journals (Sweden)

Karla M. Gregorio-Jauregui

2012-01-01

Full Text Available Preparation of magnetic nanoparticles coated with chitosan in one step by the coprecipitation method in the presence of different chitosan concentrations is reported here. Obtaining of magnetic superparamagnetic nanoparticles was confirmed by X-ray diffraction and magnetic measurements. Scanning transmission electron microscopy allowed to identify spheroidal nanoparticles with around 10-11 nm in average diameter. Characterization of the products by Fourier transform infrared spectroscopy demonstrated that composite chitosan-magnetic nanoparticles were obtained. Chitosan content in obtained nanocomposites was estimated by thermogravimetric analysis. The nanocomposites were tested in Pb2+ removal from a PbCl2 aqueous solution, showing a removal efficacy up to 53.6%. This work provides a simple method for chitosan-coated nanoparticles obtaining, which could be useful for heavy metal ions removal from water.

Blood Compatibility of Sulfonated Cladophora Nanocellulose Beads

Directory of Open Access Journals (Sweden)

Igor Rocha

2018-03-01

Full Text Available Sulfonated cellulose beads were prepared by oxidation of Cladophora nanocellulose to 2,3-dialdehyde cellulose followed by sulfonation using bisulfite. The physicochemical properties of the sulfonated beads, i.e., high surface area, high degree of oxidation, spherical shape, and the possibility of tailoring the porosity, make them interesting candidates for the development of immunosorbent platforms, including their application in extracorporeal blood treatments. A desired property for materials used in such applications is blood compatibility; therefore in the present work, we investigate the hemocompatibility of the sulfonated cellulose beads using an in vitro whole blood model. Complement system activation (C3a and sC5b-9 levels, coagulation activation (thrombin-antithrombin (TAT levels and hemolysis were evaluated after whole blood contact with the sulfonated beads and the results were compared with the values obtained with the unmodified Cladophora nanocellulose. Results showed that neither of the cellulosic materials presented hemolytic activity. A marked decrease in TAT levels was observed after blood contact with the sulfonated beads, compared with Cladophora nanocellulose. However, the chemical modification did not promote an improvement in Cladophora nanocellulose hemocompatibility in terms of complement system activation. Even though the sulfonated beads presented a significant reduction in pro-coagulant activity compared with the unmodified material, further modification strategies need to be investigated to control the complement activation by the cellulosic materials.

Blood Compatibility of Sulfonated Cladophora Nanocellulose Beads.

Science.gov (United States)

Rocha, Igor; Lindh, Jonas; Hong, Jaan; Strømme, Maria; Mihranyan, Albert; Ferraz, Natalia

2018-03-07

Sulfonated cellulose beads were prepared by oxidation of Cladophora nanocellulose to 2,3-dialdehyde cellulose followed by sulfonation using bisulfite. The physicochemical properties of the sulfonated beads, i.e., high surface area, high degree of oxidation, spherical shape, and the possibility of tailoring the porosity, make them interesting candidates for the development of immunosorbent platforms, including their application in extracorporeal blood treatments. A desired property for materials used in such applications is blood compatibility; therefore in the present work, we investigate the hemocompatibility of the sulfonated cellulose beads using an in vitro whole blood model. Complement system activation (C3a and sC5b-9 levels), coagulation activation (thrombin-antithrombin (TAT) levels) and hemolysis were evaluated after whole blood contact with the sulfonated beads and the results were compared with the values obtained with the unmodified Cladophora nanocellulose. Results showed that neither of the cellulosic materials presented hemolytic activity. A marked decrease in TAT levels was observed after blood contact with the sulfonated beads, compared with Cladophora nanocellulose. However, the chemical modification did not promote an improvement in Cladophora nanocellulose hemocompatibility in terms of complement system activation. Even though the sulfonated beads presented a significant reduction in pro-coagulant activity compared with the unmodified material, further modification strategies need to be investigated to control the complement activation by the cellulosic materials.

Biosensing based on magnetically induced self-assembly of particles in magnetic colloids.

Science.gov (United States)

Yang, Ye; Morimoto, Yoshitaka; Takamura, Tsukasa; Sandhu, Adarsh

2012-03-01

Superparamagnetic beads and nonmagnetic beads of different sizes were assembled to form a "ring-structure" in a magnetorheological (MR) fluid solution by the application of external magnetic fields. For superparamagnetic beads and non-magnetic beads functionalized with probe and target molecules, respectively, the ring-structure was maintained even after removing the external magnetic field due to biomolecular bonding. Several experiments are described, including the formation process of ring-structures with and without molecular interactions, the accelerating effect of external magnetic fields, and the effect of biotin concentration on the structures of the rings. We define the small nonmagnetic particles as "petals" because the whole structure looks like a flower. The number of remnant ring petals was a function of the concentration of target molecules in the concentration range of 0.0768 ng/ml-3.8419 ng/ml which makes this protocol a promising method for biosensing. Not only was the formation process rapid, but the resulting two-dimensional colloidal system also offers a simple method for reducing reagent consumption and waste generation.

Ecohydraulics of Strings and Beads in Bedrock Rivers

Science.gov (United States)

Wohl, E.

2016-12-01

Twenty years ago, Jack Stanford and others described rivers in bedrock canyons as resembling beads on a string when viewed in planform. The beads are relatively wide, low gradient river segments with floodplains, whereas the strings are the intervening steep, narrow river segments with minimal floodplain development. This pattern of longitudinal variations in channel and valley morphology along bedrock canyon rivers is very common, from small channels to major rivers such as the Colorado. Basic understanding of river ecosystems, as well as limited studies, indicates that the beads are more retentive and biologically productive. Although both strings and beads can provide habitat for diverse organisms, strings are more likely to serve as migration corridors, whereas beads provide spawning and nursery habitat, facilitate lateral (channel-floodplain) and vertical (channel-hyporheic) exchanges and associated habitat diversity, and retain dissolved and particulate organic matter. Recognition of the different characteristics and functions of strings and beads can be used to identify their spatial distribution along a river or within a river network and the hydraulically driven processes that sustain channel form, water quality, and biota within strings and beads. Diverse modeling approaches can then be used to quantify the fluxes of water and sediment needed to maintain these hydraulically driven processes. This conceptual framework is illustrated using examples from mountain streams in the Southern Rockies and canyon rivers in the southwestern United States.

Study on the etched carnelian beads unearthed in China

Institute of Scientific and Technical Information of China (English)

Deyun Zhao

2014-01-01

Etched carnelian beads originated in the Indus Civilization;this kind of ornaments and its manufacturing techniques were spread to the whole Eurasia Continent.The etched carnelian beads unearthed in China can be classified into four types,the comparisons of which to their foreign counterparts may reveal their different sources and diffusion routes.The etched carnelian beads and their glass imitations unearthed in China had influences to the making of the glass "eye beads" in

(Au/PANA/PVAc) nanofibers as a novel composite matrix for albumin and streptavidin immobilization

Energy Technology Data Exchange (ETDEWEB)

Golshaei, Rana [University of Kashan, Institute of Nano Science and Nano Technology, Kashan, P.O. Box 87317-51167, Islamic Republic of Iran (Iran, Islamic Republic of); Guler, Zeliha [Istanbul Technical University, Nanoscience and Nanoengineering, Maslak, Istanbul 34469 (Turkey); Sarac, Sezai A., E-mail: sarac@itu.edu.tr [Istanbul Technical University, Nanoscience and Nanoengineering, Maslak, Istanbul 34469 (Turkey); Istanbul Technical University, Department of Chemistry and Polymer Science and Technology, Maslak, Istanbul 34469 (Turkey)

2016-03-01

A novel electrospun nanofiber mat (Au/PANA/PVAc) consists of (Gold/Poly Anthranilic acid) (Au/PANA) core/shell nanostructures as a support material for protein immobilization that was developed and characterized by electrochemical impedance spectroscopy. In the core/shells, PANA served carboxyl groups (− COOH) for covalent protein immobilization and Au enhanced the electrochemical properties by acting as tiny conduction centers to facilitate electron transfer. Covalent immobilization of albumin and streptavidin as model proteins onto the (Au/PANA/PVAc) nanofibers was carried out by using 1-ethyl-3-(dimethyl-aminopropyl) carbodiimide hydrochloride (EDC)/N-hydroxyl succinimide (NHS) activation. PVAc nanofibers were compared with Au/PANA/PVAc nanofibers before and after protein immobilization. The successful covalent binding of both albumin and streptavidin onto (Au/PANA/PVAc) nanofibers was confirmed by FTIR-ATR, Electron Microscopy/Energy-Dispersive X-ray Spectroscopy SEM/EDX and Electrochemical impedance spectroscopy (EIS). The nanofibers became resistive due to protein immobilization and the higher charge transfer resistance was observed after higher amount of protein was immobilized. - Highlights: • Au/PANA/PVAc nanofibers with (COOH) groups as a suitable supports for covalent immobilization of proteins. • Increasing of the resistivity of the nanofibers after immobilization of the proteins. • Activation of Au/PANA/PVAc nanofibers by using EDC/NHS.

Inhibitive Effect of antibiotic-loaded beads to cure chronic osteomyelitis in developing country : Hand-made vs commercial beads

NARCIS (Netherlands)

Rasyid, Hcrmawan N.; Van Horn, Jim R.; Van der Mei, Henny C.; Soegijoko, Sooegijardjo; Busscher, Henk J.; Neut, Danielle; Ibrahim, F; Osman, NAA; Usman, J; Kadri, NA

2007-01-01

Local antibiotic-loaded beads have been approved for standard treatment of orthopaedic pathogens, especially chronic osteomyelitis. Septopal (R), the only commercial local antibiotic bead available on the market, is expensive and contains only gentamicin. This study aimed to compare the in vitro

Configurational Statistics of Magnetic Bead Detection with Magnetoresistive Sensors

DEFF Research Database (Denmark)

Henriksen, Anders Dahl; Ley, Mikkel Wennemoes Hvitfeld; Flyvbjerg, Henrik

2015-01-01

Magnetic biosensors detect magnetic beads that, mediated by a target, have bound to a functionalized area. This area is often larger than the area of the sensor. Both the sign and magnitude of the average magnetic field experienced by the sensor from a magnetic bead depends on the location...... of the bead relative to the sensor. Consequently, the signal from multiple beads also depends on their locations. Thus, a given coverage of the functionalized area with magnetic beads does not result in a given detector response, except on the average, over many realizations of the same coverage. We present...... a systematic theoretical analysis of how this location-dependence affects the sensor response. The analysis is done for beads magnetized by a homogeneous in-plane magnetic field. We determine the expected value and standard deviation of the sensor response for a given coverage, as well as the accuracy...

Molecular beacon based biosensor for the sequence-specific detection of DNA using DNA-capped gold nanoparticles-streptavidin conjugates for signal amplification

International Nuclear Information System (INIS)

Fang, Xian; Jiang, Wei; Han, Xiaowei; Zhang, Yuzhong

2013-01-01

We describe a highly sensitive and selective molecular beacon-based electrochemical impedance biosensor for the sequence-specific detection of DNA. DNA-capped conjugates between gold nanoparticles (Au-NPs) and streptavidin are used for signal amplification. The molecular beacon was labeled with a thiol at its 5′ end and with biotin at its 3′ end, and then immobilized on the surface of a bare gold electrode through the formation of Au-S bonds. Initially, the molecular beacon is present in the “closed” state, and this shields the biotin from being approached by streptavidin due to steric hindrance. In the presence of the target DNA, the target DNA molecules hybridize with the loop and cause a conformational change that moves the biotin away from the surface of the electrode. The biotin thereby becomes accessible for the reporter (the DNA-streptavidin capped Au-NPs), and this results in a distinct increase in electron transfer resistance. Under optimal conditions, the increase in resistance is linearly related to the logarithm of the concentration of complementary target DNA in the range from 1.0 fM to 0.1 μM, with a detection limit of 0.35 fM (at an S/N of 3). This biosensor exhibits good selectivity, and acceptable stability and reproducibility. (author)

Calibration beads containing luminescent lanthanide ion complexes

Science.gov (United States)

The reliability of lanthanide luminescence measurements, by both flow cytometry and digital microscopy, will be enhanced by the availability of narrow-band emitting lanthanide calibration beads. These beads can also be used to characterize spectrographic instruments, including mi...

[Evaluation of TB-beads assay utilizing the technique of magnetic beads--an innovative assay method for detection of acid fast bacilli].

Science.gov (United States)

Ohkuma, Masanori; Ikeda, Katsuyoshi; Obayashi, Konen; Ando, Yukio; Koriyama, Toyoyasu; Kimu, Minhi; Hirose, Nobuyuki; Nagasawa, Zenzo; Miyamoto, Hiroshi

2012-01-01

The centrifuge method with the use of Semi-Alkalin Proteinase (SAP) and NALC-NaOH, recommended by the "2007 edition of the assay guideline for detection of Mycobacterium tuberculosis," has significantly contributed to improving the sensitivities and specificities of both smear and culture tests for detection of acid fast bacilli (AFB). However, this method poses some challenges in terms of its cumbersome and time-consuming assay protocol. "TB-beads (Kyokuto Pharmaceutical Industrial Co., Ltd.)" is a newly-developed method for detection of AFB utilizing magnetic beads. We evaluated the quality of this method in comparison with the centrifuge method, focusing on the results of smear and culture tests. This evaluation study was conducted using both 5 positive and 5 negative sputum samples. The sensitivity of TB-beads for fluorescent smear tests, conducted using "Acri-stain," was almost the same as that of the centrifuge method. One advantage of TB-beads, however, was that it was very convenient to practice microscopic observation due to the clear background of the smeared glass slides. The comparison of the contamination rates between the two methods showed that TB-beads suggested significantly lower contamination rates. The centrifuge method resulted in 50% and 60% of contamination rates for HK Semisolid Isolation Medium and BacT/ALERT MP, respectively. On the other hand, the contamination rates of TB-beads for both of the culture methods were only 10%. With regard to the 5 positive sputum samples, the comparison of the detection rates between the centrifuge and TB-Beads method was made utilizing Myco Acid, Ogawa K, and BacT/ALERT MP. The TB-Beads method suggested higher detection rates for Myco Acid and Ogawa K, while there were no significant differences between the two methods for BacT/ALERT MP (16-23 days). TB-beads is an easy method that allows to simplify the process of smear tests, and contributes to significantly reducing the contamination rate of culture

Rapid microwave-assisted synthesis of dextran-coated iron oxide nanoparticles for magnetic resonance imaging

International Nuclear Information System (INIS)

Osborne, Elizabeth A; Atkins, Tonya M; Kauzlarich, Susan M; Gilbert, Dustin A; Liu Kai; Louie, Angelique Y

2012-01-01

Currently, magnetic iron oxide nanoparticles are the only nanosized magnetic resonance imaging (MRI) contrast agents approved for clinical use, yet commercial manufacturing of these agents has been limited or discontinued. Though there is still widespread demand for these particles both for clinical use and research, they are difficult to obtain commercially, and complicated syntheses make in-house preparation unfeasible for most biological research labs or clinics. To make commercial production viable and increase accessibility of these products, it is crucial to develop simple, rapid and reproducible preparations of biocompatible iron oxide nanoparticles. Here, we report a rapid, straightforward microwave-assisted synthesis of superparamagnetic dextran-coated iron oxide nanoparticles. The nanoparticles were produced in two hydrodynamic sizes with differing core morphologies by varying the synthetic method as either a two-step or single-step process. A striking benefit of these methods is the ability to obtain swift and consistent results without the necessity for air-, pH- or temperature-sensitive techniques; therefore, reaction times and complex manufacturing processes are greatly reduced as compared to conventional synthetic methods. This is a great benefit for cost-effective translation to commercial production. The nanoparticles are found to be superparamagnetic and exhibit properties consistent for use in MRI. In addition, the dextran coating imparts the water solubility and biocompatibility necessary for in vivo utilization. (paper)

Rapid microwave-assisted synthesis of dextran-coated iron oxide nanoparticles for magnetic resonance imaging.

Science.gov (United States)

Osborne, Elizabeth A; Atkins, Tonya M; Gilbert, Dustin A; Kauzlarich, Susan M; Liu, Kai; Louie, Angelique Y

2012-06-01

Currently, magnetic iron oxide nanoparticles are the only nanosized magnetic resonance imaging (MRI) contrast agents approved for clinical use, yet commercial manufacturing of these agents has been limited or discontinued. Though there is still widespread demand for these particles both for clinical use and research, they are difficult to obtain commercially, and complicated syntheses make in-house preparation unfeasible for most biological research labs or clinics. To make commercial production viable and increase accessibility of these products, it is crucial to develop simple, rapid and reproducible preparations of biocompatible iron oxide nanoparticles. Here, we report a rapid, straightforward microwave-assisted synthesis of superparamagnetic dextran-coated iron oxide nanoparticles. The nanoparticles were produced in two hydrodynamic sizes with differing core morphologies by varying the synthetic method as either a two-step or single-step process. A striking benefit of these methods is the ability to obtain swift and consistent results without the necessity for air-, pH- or temperature-sensitive techniques; therefore, reaction times and complex manufacturing processes are greatly reduced as compared to conventional synthetic methods. This is a great benefit for cost-effective translation to commercial production. The nanoparticles are found to be superparamagnetic and exhibit properties consistent for use in MRI. In addition, the dextran coating imparts the water solubility and biocompatibility necessary for in vivo utilization.

Solid-phase cloning for high-throughput assembly of single and multiple DNA parts

DEFF Research Database (Denmark)

Lundqvist, Magnus; Edfors, Fredrik; Sivertsson, Åsa

2015-01-01

We describe solid-phase cloning (SPC) for high-throughput assembly of expression plasmids. Our method allows PCR products to be put directly into a liquid handler for capture and purification using paramagnetic streptavidin beads and conversion into constructs by subsequent cloning reactions. We ...

Discrete dipole approximation simulation of bead enhanced diffraction grating biosensor

International Nuclear Information System (INIS)

Arif, Khalid Mahmood

2016-01-01

We present the discrete dipole approximation simulation of light scattering from bead enhanced diffraction biosensor and report the effect of bead material, number of beads forming the grating and spatial randomness on the diffraction intensities of 1st and 0th orders. The dipole models of gratings are formed by volume slicing and image processing while the spatial locations of the beads on the substrate surface are randomly computed using discrete probability distribution. The effect of beads reduction on far-field scattering of 632.8 nm incident field, from fully occupied gratings to very coarse gratings, is studied for various bead materials. Our findings give insight into many difficult or experimentally impossible aspects of this genre of biosensors and establish that bead enhanced grating may be used for rapid and precise detection of small amounts of biomolecules. The results of simulations also show excellent qualitative similarities with experimental observations. - Highlights: • DDA was used to study the relationship between the number of beads forming gratings and ratio of first and zeroth order diffraction intensities. • A very flexible modeling program was developed to design complicated objects for DDA. • Material and spatial effects of bead distribution on surfaces were studied. • It has been shown that bead enhanced grating biosensor can be useful for fast detection of small amounts of biomolecules. • Experimental results qualitatively support the simulations and thus open a way to optimize the grating biosensors.

Microarray of DNA probes on carboxylate functional beads surface

Institute of Scientific and Technical Information of China (English)

黄承志; 李原芳; 黄新华; 范美坤

2000-01-01

The microarray of DNA probes with 5’ -NH2 and 5’ -Tex/3’ -NH2 modified terminus on 10 um carboxylate functional beads surface in the presence of 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide (EDC) is characterized in the preseni paper. it was found that the microarray capacity of DNA probes on the beads surface depends on the pH of the aqueous solution, the concentra-tion of DNA probe and the total surface area of the beads. On optimal conditions, the minimum distance of 20 mer single-stranded DNA probe microarrayed on beads surface is about 14 nm, while that of 20 mer double-stranded DNA probes is about 27 nm. If the probe length increases from 20 mer to 35 mer, its microarray density decreases correspondingly. Mechanism study shows that the binding mode of DNA probes on the beads surface is nearly parallel to the beads surface.

Microarray of DNA probes on carboxylate functional beads surface

Institute of Scientific and Technical Information of China (English)

无

2000-01-01

The microarray of DNA probes with 5′-NH2 and 5′-Tex/3′-NH2 modified terminus on 10 m m carboxylate functional beads surface in the presence of 1-ethyl-3-(3-dimethylaminopropyl)- carbodiimide (EDC) is characterized in the present paper. It was found that the microarray capacity of DNA probes on the beads surface depends on the pH of the aqueous solution, the concentration of DNA probe and the total surface area of the beads. On optimal conditions, the minimum distance of 20 mer single-stranded DNA probe microarrayed on beads surface is about 14 nm, while that of 20 mer double-stranded DNA probes is about 27 nm. If the probe length increases from 20 mer to 35 mer, its microarray density decreases correspondingly. Mechanism study shows that the binding mode of DNA probes on the beads surface is nearly parallel to the beads surface.

Effects of agitation rate on aggregation during beads-to-beads subcultivation of microcarrier culture of human mesenchymal stem cells.

Science.gov (United States)

Takahashi, Iori; Sato, Keigo; Mera, Hisashi; Wakitani, Shigeyuki; Takagi, Mutsumi

2017-06-01

With the aim to utilize human mesenchymal stem cells (hMSCs) grown in large scale for regenerative medicine, effects of agitation rate on aggregation during beads-to-beads subcultivation of microcarrier culture of hMSCs were studied. hMSCs could attach and grew on surface-type microcarriers of Cytodex 1, whereas almost no cell elongation and growth were observed on porous type microcarriers of Cytopores. The percentages of aggregated Cytodex 1 microcarriers at an agitation rate of 60 and 90 rpm were lower than that at 30 rpm, which was the lowest agitation rate necessary for the suspension of Cytodex 1 microcarriers, and the cells grew fastest at 60 rpm. hMSC could be subcultivated on Cytodex 1 by the beads-to-beads method at both 30 and 60 rpm without trypsinization. However, agitation at 60 rpm resulted in a markedly lower percentage of aggregated microcarriers not only before but also after subcultivation. The percentages of CD90- and CD166-positive cells among cells grown on Cytodex 1 at 60 rpm (91.5 and 87.6 %) were comparable to those of cells grown in the pre-culture on dishes. In conclusion, hMSCs could be subcultivated on Cytodex 1 by beads-to-beads method maintaining the expressions of the cell surface antigens CD90 and CD166, while adjusting agitation rate could decrease the microcarrier aggregation.

Process for a preparing a coating composition. [electrom irradiation

Energy Technology Data Exchange (ETDEWEB)

Kimura, T; Harada, H; Kobayashi, J; Nakamoto, H; Sunano, K

1968-07-16

An easily hardenable acrylic coating composition is prepared with low energy electron beams to develop a surface coating process without requiring solvents, and which may be widely applied by industry. The process comprises dissolving a polymer with a molecular weight in the 5,000 to 500,000 range in a monomer consisting of at least 30% by weight of acrylic monomer and 70% by weight of other vinyl monomers. The polymer is obtained by the polymerization of 1 to 40% by weight of vinyl monomer containing carboxyl radicals, 30 to 99% by weight of methacrylic monomer and 0 to 69% by weight of other copolymerizable vinyl monomers. Then, one mole of carboxyl radical of the solution reacts with 0.1 to 1.0 mole of vinyl monomer containing a glycidyl radical. In an embodiment, 17.5% by weight of methacrylate are dissolved in 82.5% of alkyl acrylate and undergo suspension polymerization in water in the presence of a catalyst to produce a beads-like polymer of molecular weight in the 5,000 to 500,000 range. Thereafter, 120 parts of the beads-like polymer are dissolved in 180 parts of acrylic monomer in the presence of a polymerization inhibitor. To this solution are added 22 parts of glycidyl methacrylate to react with carboxyl radicals, thereby obtaining non-solvent coating materials which contain the side chain vinyl radicals in the polymer. The acceleration voltage of the electron beams employed in the polymerization is generally 0.1 to 2.0 MeV. The dose rate to harden the coatings is in the range of 0.1 to 2.0 Mrad/sec.

Enteric coated HPMC capsules plugged with 5-FU loaded microsponges: a potential approach for treatment of colon cancer

Directory of Open Access Journals (Sweden)

Ankita Gupta

2015-09-01

Full Text Available The work was aimed at developing novel enteric coated HPMC capsules (ECHC plugged with 5 Florouracil (5-FU loaded Microsponges in combination with calcium pectinate beads. Modified quasi-emulsion solvent diffusion method was used to formulate microsponges based on 32 factorial design and the effects of independent variables (volume of organic solvent and Eudragit RS100 content on the dependent variables (Particle size, %EE & % CDR were determined. The optimized microsponges (F4 were characterized by SEM, PXRD, TGA and were plugged along with calcium pectinate beads in HPMC capsules and the HPMC capsules were further coated with enteric polymer Eudragit L 100 (Ed-L100 and/ or Eudrgit S 100 (Ed-S 100 in different proportions. In vitro release study of ECHC was performed in various release media sequentially SGF for 2 h, followed by SIF for the next 6 h and then in SCF (in the presence and absence of pectinase enzyme for further 16 h. Drug release was retarded on coating with EdS-100 in comparison to blend of EdS-100: EdL-100 coating. The percentage of 5-FU released at the end of 24 h from ECHC 3 was 97.83 ± 0.12% in the presence of pectinase whereas in control study it was 40.08 ± 0.02% drug. The optimized formulation was subjected to in vivo Roentgenographic studies in New Zealand white rabbits to analyze the in vivo behavior of the developed colon targeted capsules. Pharmacokinetic studies in New Zealand white rabbits were conducted to determine the extent of systemic exposure provided by the developed formulation in comparison to 5-FU aqueous solutions. Thus, enteric coated HPMC capsules plugged with 5-FU loaded microsponges and calcium pectinate beads proved to be promising dosage form for colon targeted drug delivery to treat colorectal cancer.

Permanent magnet system to guide superparamagnetic particles

Science.gov (United States)

Baun, Olga; Blümler, Peter

2017-10-01

A new concept of using permanent magnet systems for guiding superparamagnetic nano-particles on arbitrary trajectories over a large volume is proposed. The basic idea is to use one magnet system which provides a strong, homogeneous, dipolar magnetic field to magnetize and orient the particles, and a second constantly graded, quadrupolar field, superimposed on the first, to generate a force on the oriented particles. In this configuration the motion of the particles is driven predominantly by the component of the gradient field which is parallel to the direction of the homogeneous field. As a result, particles are guided with constant force and in a single direction over the entire volume. The direction is simply adjusted by varying the angle between quadrupole and dipole. Since a single gradient is impossible due to Gauß' law, the other gradient component of the quadrupole determines the angular deviation of the force. However, the latter can be neglected if the homogeneous field is stronger than the local contribution of the quadrupole field. A possible realization of this idea is a coaxial arrangement of two Halbach cylinders. A dipole to evenly magnetize and orient the particles, and a quadrupole to generate the force. The local force was calculated analytically for this particular geometry and the directional limits were analyzed and discussed. A simple prototype was constructed to demonstrate the principle in two dimensions on several nano-particles of different size, which were moved along a rough square by manual adjustment of the force angle. The observed velocities of superparamagnetic particles in this prototype were always several orders of magnitude higher than the theoretically expected value. This discrepancy is attributed to the observed formation of long particle chains as a result of their polarization by the homogeneous field. The magnetic moment of such a chain is then the combination of that of its constituents, while its hydrodynamic radius

Half bead welding technique

International Nuclear Information System (INIS)

Canonico, D.A.; Holz, P.P.

1978-05-01

The ORNL has employed the Section XI half-bead procedure for six repair welds. Table 2 identifies the repairs and the components upon which they were accomplished. The weld repairs were performed to permit us to evaluate material properties, residual stresses, weld repair procedures, and structural behavior of repaired pressure vessels. As a consequence of our study we concluded that when the half bead procedure is correctly applied: (1) there is no metallurgical degradation of the base material, (2) residual stresses of yield point magnitude will be present, and (3) the structural integrity of the pressure vessel is not impaired at Charpy V-notch upper shelf temperatures

Moessbauer studies of superparamagnetic ferrite nanoparticles for functional application

Energy Technology Data Exchange (ETDEWEB)

Mazeika, K., E-mail: kestas@ar.fi.lt; Jagminas, A.; Kurtinaitiene, M. [SSRI Center for Physical Sciences and Technology (Lithuania)

2013-04-15

Nanoparticles of CoFe{sub 2}O{sub 4} and MnFe{sub 2}O{sub 4} prepared for functional applications in nanomedicine were studied using Moessbauer spectrometry. Superparamagnetic properties of nanoparticles of different size and composition were compared applying collective excitations and multilevel models for the description of the Moessbauer spectra.

Synthesis and characterization of cationic lipid coated magnetic nanoparticles using multiple emulsions as microreactors

Science.gov (United States)

Akbaba, Hasan; Karagöz, Uğur; Selamet, Yusuf; Kantarcı, A. Gülten

2017-03-01

The aim of this study was to develop a novel iron oxide nanoparticle synthesis method with in-situ surface coating. For this purpose multiple emulsions were used as microreactors for the first time and magnetic iron oxide particles synthesized in the core of cationic solid lipid nanoparticles. DLS, SEM, TEM, VSM, Raman Spectrometer, XRD, and XPS techniques were performed for characterization of the magnetic nanoparticles. Obtained magnetic nanoparticles are superparamagnetic and no additional process was needed for surface adjustments. They are positively charged as a result of cationic lipid coating and has appropriate particle size (<30 nm) for drug or nucleic acid delivery. Structure analysis showed that magnetic core material is in the form of magnetite. Saturation magnetization value was measured as 15-17 emu g-1 for lipid coated magnetic nanoparticles obtained by multiple emulsion method which is reasonably sufficient for magnetic targeting.

Splenic red pulp macrophages are intrinsically superparamagnetic and contaminate magnetic cell isolates.

Science.gov (United States)

Franken, Lars; Klein, Marika; Spasova, Marina; Elsukova, Anna; Wiedwald, Ulf; Welz, Meike; Knolle, Percy; Farle, Michael; Limmer, Andreas; Kurts, Christian

2015-08-11

A main function of splenic red pulp macrophages is the degradation of damaged or aged erythrocytes. Here we show that these macrophages accumulate ferrimagnetic iron oxides that render them intrinsically superparamagnetic. Consequently, these cells routinely contaminate splenic cell isolates obtained with the use of MCS, a technique that has been widely used in immunological research for decades. These contaminations can profoundly alter experimental results. In mice deficient for the transcription factor SpiC, which lack red pulp macrophages, liver Kupffer cells take over the task of erythrocyte degradation and become superparamagnetic. We describe a simple additional magnetic separation step that avoids this problem and substantially improves purity of magnetic cell isolates from the spleen.

Sensitive detection of point mutation by electrochemiluminescence and DNA ligase-based assay

Science.gov (United States)

Zhou, Huijuan; Wu, Baoyan

2008-12-01

The technology of single-base mutation detection plays an increasingly important role in diagnosis and prognosis of genetic-based diseases. Here we reported a new method for the analysis of point mutations in genomic DNA through the integration of allele-specific oligonucleotide ligation assay (OLA) with magnetic beads-based electrochemiluminescence (ECL) detection scheme. In this assay the tris(bipyridine) ruthenium (TBR) labeled probe and the biotinylated probe are designed to perfectly complementary to the mutant target, thus a ligation can be generated between those two probes by Taq DNA Ligase in the presence of mutant target. If there is an allele mismatch, the ligation does not take place. The ligation products are then captured onto streptavidin-coated paramagnetic beads, and detected by measuring the ECL signal of the TBR label. Results showed that the new method held a low detection limit down to 10 fmol and was successfully applied in the identification of point mutations from ASTC-α-1, PANC-1 and normal cell lines in codon 273 of TP53 oncogene. In summary, this method provides a sensitive, cost-effective and easy operation approach for point mutation detection.

Doxorubicin loaded PVA coated iron oxide nanoparticles for targeted drug delivery

International Nuclear Information System (INIS)

Kayal, S.; Ramanujan, R.V.

2010-01-01

Magnetic drug targeting is a drug delivery system that can be used in locoregional cancer treatment. Coated magnetic particles, called carriers, are very useful for delivering chemotherapeutic drugs. Magnetic carriers were synthesized by coprecipitation of iron oxide followed by coating with polyvinyl alcohol (PVA). Characterization was carried out using X-ray diffraction, TEM, TGA, FTIR and VSM techniques. The magnetic core of the carriers was magnetite (Fe 3 O 4 ), with average size of 10 nm. The room temperature VSM measurements showed that magnetic particles were superparamagnetic. The amount of PVA bound to the iron oxide nanoparticles were estimated by thermogravimetric analysis (TGA) and the attachment of PVA to the iron oxide nanoparticles was confirmed by FTIR analysis. Doxorubicin (DOX) drug loading and release profiles of PVA coated iron oxide nanoparticles showed that up to 45% of adsorbed drug was released in 80 h, the drug release followed the Fickian diffusion-controlled process. The binding of DOX to the PVA was confirmed by FTIR analysis. The present findings show that DOX loaded PVA coated iron oxide nanoparticles are promising for magnetically targeted drug delivery.

Facile Synthesis Polyethylene Glycol Coated Magnetite Nanoparticles for High Colloidal Stability

Directory of Open Access Journals (Sweden)

Mun Foong Tai

2016-01-01

Full Text Available Polyethylene glycol (PEG is one of the most frequently used synthetic polymers for surface modifications of magnetite nanoparticles (MNPs to provide a new opportunity for constructing high colloidal stability. Herein, a facile in situ coprecipitation technique is described for the synthesis of PEG coated MNPs using ammonium hydroxide as the precipitating agent. The structure and morphology of the prepared PEG coated MNPs samples were characterized by Fourier transform infrared (FTIR spectroscopy, X-ray spectroscopy, thermogravimetric analysis (TGA, and the high resolution transmission electron microscopy (HRTEM. In this study, all samples demonstrated hydrodynamic size in the range of 32 to 43 nm with narrow size distribution. In addition, the magnetic properties of resultant samples were investigated using a vibrating sample magnetometer (VSM to reveal the superparamagnetic behaviour with saturation magnetization. The saturation magnetization of PEG coated MNPs samples was in the range of 63 to 66 emu/g at 300 K. Interestingly, it was found that 1.0 g of PEG coated MNPs exhibited high colloidal stability in a basic solution (pH = 10 and nitrile (NBR latex up to 21 days as compared to the unmodified MNPs during the sedimentation test.

Harmonic decomposition of magneto-optical signal from superparamagnetic Fe3O4 nanoparticles

Science.gov (United States)

Syed, Maarij; Patterson, Cody; Takemura, Yasushi

Superparamagnetic nanoparticles (SPNPs) are expected to play an increasingly important role in bio-imaging and therapy. These applications rely on understanding SPNPs magnetic properties which have been successfully characterized by AC Faraday rotation (FR). AC FR is used here to build on results presented earlier by measuring solutions of surfactant-coated magnetite nanoparticles. The setup employs a He-Ne laser, polarizing components, a sinusoidal B-field, and a lock-in detection scheme to measure the SPNPs FR. Such a setup provides a novel, economical way of determining important magnetic properties of SPNPs. The main intensity signal (1f) along with higher harmonics are collected and analyzed to calculate quantities such as the Verdet constant and the magnetic moment. We hope further analysis can also reveal details of size distribution and relaxation times of SPNPs. We will present results from samples with various concentrations as well as a particular concentration subjected to a range of B-field frequencies (between 800 Hz and 14 kHz). Findings are compared to results from more traditional techniques like magnetic susceptibility measurements, magnetic force microscopy, etc. We will also address the comparative advantages of this technique and its limitations.

Magnetite Nanoparticles Coated with Rifampicin and Chlortetracycline for Drug Delivery Applications

International Nuclear Information System (INIS)

Nadejde, Claudia; Ciurlica, Ecaterina Foca-nici; Creanga, Dorina; Carlescu, Aurelian; Badescu, Vasile

2010-01-01

Four types of biocompatible magnetic fluids based on superparamagnetic nanoparticles with Fe 3 O 4 cores were functionalized with antibiotics (rifampicin or chlortetracycline) as potential candidates for in vivo biomedical applications, such as magnetically controlled drug delivery. The synthesis consisted in coprecipitation of iron oxide in basic, as well as in acid medium, followed by the dispersion of the resulted magnetite nanoparticles in aqueous solution containing the antibiotic. The chosen method to prepare the magnetite-core/drug-shell systems avoided intermediate organic coating of the magnetic nanoparticles. Comparative analysis of the rheological features of the aqueous magnetic fluid samples was performed. The structural features of the coated magnetic particles were investigated by X-Ray Diffraction (XRD), Transmission Electron Microscopy (TEM) and Vibrating Sample Magnetometry (VSM). Good crystallinity and adequate stability in time were evidenced. Drug delivery curves were spectrophotometrically provided.

Electroactivity of avidin and streptavidin. Avidin signals at mercury and carbon electrodes respond to biotin binding

Czech Academy of Sciences Publication Activity Database

Havran, Luděk; Billová, Sabina; Paleček, Emil

2004-01-01

Roč. 16, 13-14 (2004), s. 1139-1148 ISSN 1040-0397 R&D Projects: GA ČR GA204/03/0566; GA MŠk OC D21.002; GA AV ČR KJB4004302 Institutional research plan: CEZ:AV0Z5004920 Keywords : streptavidin * avidin-biotin interaction * electrochemical methods Subject RIV: BO - Biophysics Impact factor: 2.038, year: 2004

Comparative evaluation of various solid phases for the development of coated tube assays for the estimation of progesterone in human serum, bovine serum and bovine milk

International Nuclear Information System (INIS)

Karir, Tarveen; Samuel, Grace; Sivaprasad, N.; Venkatesh, Meera

2009-01-01

Immobilization of progesterone antibody using three polystyrene surfaces and two progesterone radiotracers for use in the development of a coated tube assay for the evaluation of progesterone levels in human serum, bovine serum and bovine milk was studied. The selection of the solid phase and the tracers were based on the maximum binding, non-specific binding, sensitivity and percentage recovery. Amongst the polystyrene tubes studied, streptavidin coated tubes showed the acceptable assay features such as low non-specific binding (0.5-1.0%), adequate sensitivity (0.13-0.16 ng/ml) and recovery (85-115%) for all the three sample matrices, human serum, bovine serum and bovine milk.

Characterizing and quantifying superparamagnetic magnetite particles in serpentinized mantle peridotite observed in continental ophiolite complexes.

Science.gov (United States)

Ortiz, E.; Vento, N. F. R.; Tominaga, M.; Beinlich, A.; Einsle, J. F.; Buisman, I.; Ringe, E.; Schrenk, M. O.; Cardace, D.

2017-12-01

Serpentinization of mantle peridotite has been recognized as one of the most important energy factories for the deep biosphere. To better evaluate the habitability of the deep biosphere, it is crucial to understand the link between in situ peridotite serpentinization processes and associated magnetite and hydrogen production. Previous efforts in correlating magnetite and hydrogen production during serpentinization processes are based primarily on laboratory experiments and numerical modeling, being challenged to include the contribution of superparamagnetic-sized magnetites (i.e., extremely fine-grained magnetite, petrographically observed as a "pepper flake" like texture in many natural serpentinized rock samples). To better estimate the abundance of superparamagnetic grains, we conducted frequency-dependent susceptibility magnetic measurements at the Institute of Rock Magnetism on naturally serpentinized rock samples from the Coast Range Ophiolite Microbial Observatory (CROMO) in California, USA and the Atlin Ophiolite (British Columbia). In addition, we conducted multiscale EDS phase mapping, BackScattered Electron (BSE) scanning, FIB-nanotomography and STEM-EELS to identify and quantify the superparamagnetic minerals that contribute to the measured magnetic susceptibility signals in our rock samples. Utilizing a multidisciplinary approach, we aim to improve the estimation of hydrogen production based on the abundance of magnetite, that includes the contribution of superparamagnetic particle size magnetite, to ultimately provide a more accurate estimation of bulk deep-biomass hosted by in situ serpentinization processes.

Preparation of Bio-beads and Their Atrazine Degradation Characteristics

Institute of Scientific and Technical Information of China (English)

BI Hai-tao; ZHANG Lan-ying; LIU Na; ZHU Bo-lin

2011-01-01

Screened atrazine-mineralizing bacterium-Pseudomonas W4 was embedded inside an improved PVAH3BO3 embedment matrix to make bio-beads to degrade atrazine. The atrazine degradation characteristics were studied. The preparation procedure of bio-beads was as follows: (1) preparing a mixture of 100, 12.5, 10, 1.5 and 1 g/L PVA, bentonite(Ca), activated carbon powder, sodium alginate and centrifuged Pseudomonas W4 bacterium, respectively; (2) the mixture was dropped into a gently stirred cross linker solution(pH=6.7) and cured at 10 ℃ for 24 h.The optimal atrazine degradation conditions by bio-beads were as follows: pH=7, the auxiliary carbon source was glucose, and the concentration of glucose was greater than 325 mg/L. The bio-beads demonstrated stronger tolerance ability than the free microorganism to the increase of PCBs, hydrogen ion and hydroxide ion. SEM images show the uniform distribution of the microorganism inside bio-beads and the porous cross-linked structure of bio-beads which provides excellent mass transfer capacity.

Preparation, characterization and application of superparamagnetic iron oxide nanoparticles modified with natural polymers for removal of {sup 60}Co-radionuclides from aqueous solution

Energy Technology Data Exchange (ETDEWEB)

Sharaf El-Deen, Gehan E. [Atomic Energy Authority, Cairo (Egypt). Radioactive Waste Management Dept.; Imam, Neama G. [Atomic Energy Authority, Cairo (Egypt). Experimental Physics Dept.; Elettra, Sincrotrone, Trieste (Italy); Ayoub, Refaat R. [Atomic Energy Authority, Cairo (Egypt). Nuclear Chemistry Dept.

2017-04-01

Superparamagnetic iron oxide nanoparticles (IO-MNPs) coated with natural polymers, starch (IO-S MNPs) and dextrin (IO-D MNPs), were synthesized by modified co-precipitation method. IO and hybrid-IO-MNPs were characterized by XRD, SEM, HRTEM, FT-IR spectroscopy, vibrating sample magnetometer (VSM) and zeta potential (ZP). IO-S MNPs and IO-D MNPs have IO core-shell structure with core of 10.8 nm and 13.8 nm and shell of 7.5 nm and 5.9 nm, respectively. The efficiency of the hybrid IO-MNPs for sorption of {sup 60}Co(II)-radionuclides from aqueous solution was investigated under varying experimental conditions. Kinetic data were described well by pseudo-second-order mode, sorption isotherms were fitted quite with Freundlich model with maximum adsorption capacity 36.89 (mmol.g{sup -1})/(L.mmol{sup -1}){sup n} for IO-S MNPs and 24.9 (mmol.g{sup -1})/(L.mmol{sup -1}){sup n} for IO-D MNPs. Sorption of {sup 60}Co-radionuclides by IO-S MNPs was suppressed with salinity and most of the adsorbed {sup 60}Co onto IO-S MNPs could be remove with 0.1 M HCl solution. IO-S MNPs exhibits superparamagnetic properties, easier separation according to higher saturation magnetization (47 emu/g) and better adsorption for {sup 60}CO-radionuclides than IO-D MNPs.

Assessment of the Problems of Manual Automobile Tyre Bead ...

African Journals Online (AJOL)

The tyre-rim bead bond must be broken to carry out repairs on a failed automobile tyre. The use of the locally fabricated manual bead breaking equipment as it is being practiced today by commercial tyre repair artisans in Nigeria is characterized by drudgery. This article reports a study of the local manual bead breaking ...

Growth and morphology of thermophilic dairy starters in alginate beads.

Science.gov (United States)

Lamboley, Laurence; St-Gelais, Daniel; Champagne, Claude P; Lamoureux, Maryse

2003-06-01

The aim of this research was to produce concentrated biomasses of thermophilic lactic starters using immobilized cell technology (ICT). Fermentations were carried out in milk using pH control with cells microentrapped in alginate beads. In the ICT fermentations, beads represented 17% of the weight. Some assays were carried out with free cells without pH control, in order to compare the ICT populations with those of classical starters. With Streptococcus thermophilus, overall populations in the fermentor were similar, but maximum bead population for (8.2 x 10(9) cfu/g beads) was 13 times higher than that obtained in a traditional starter (4.9 x 10(8) cfu/ml). For both Lactobacillus helveticus strains studied, immobilized-cell populations were about 3 x 10(9) cfu/g beads. Production of immobilized Lb. bulgaricus 210R strain was not possible, since no increases in viable counts occurred in beads. Therefore, production of concentrated cell suspension in alginate beads was more effective for S. thermophilus. Photomicrographs of cells in alginate beads demonstrated that, while the morphology of S. thermophilus remained unchanged during the ICT fermentation, immobilized cells of Lb. helveticus appeared wider. In addition, cells of Lb. bulgaricus were curved and elongated. These morphological changes would also impair the growth of immobilized lactobacilli.

Binding properties of a streptavidin layer formed on a biotinylated Langmuir–Schaefer film of unfolded protein

Energy Technology Data Exchange (ETDEWEB)

Furuno, Taiji, E-mail: t_furuno@a8.keio.jp

2016-04-01

A Langmuir monolayer of carbonic anhydrase (CA) unfolded at an air/water interface was transferred onto the hydrophobic surface of a silicon wafer by means of the Langmuir–Schaefer technique. The transferred CA film was biotinylated and was incubated in a streptavidin (SAv) solution to obtain a densely packed SAv layer by biotin–SAv linkage. Biotinylated proteins including ferritin, catalase, alcohol dehydrogenase, and carbonic anhydrase were incubated with the SAv layer and binding of these proteins was examined by atomic force microscopy. High-density binding of the biotinylated proteins was observed, whereas the amount of adsorbed non-biotinylated proteins was low or negligible. The SAv layer on the Langmuir–Schaefer film of unfolded protein could become a basic architecture for protein immobilization studies. - Highlights: • Langmuir–Schaefer film of carbonic anhydrase (LSF-CA) was biotinylated. • A densely packed streptavidin (SAv) layer was formed on the biotinylated LSF-CA. • Biotinylated proteins were bound to the SAv layer at high density. • Nonspecific adsorption of intact proteins to the SAv layer was weak. • Atomic force microscopy showed the binding of proteins at molecular resolution.

Multiple functionalities of Ni nanoparticles embedded in carboxymethyl guar gum polymer: catalytic activity and superparamagnetism

International Nuclear Information System (INIS)

Sardar, Debasmita; Sengupta, Manideepa; Bordoloi, Ankur; Ahmed, Md. A.; Neogi, S.K.; Bandyopadhyay, Sudipta; Jain, Ruchi; Gopinath, Chinnakonda S.; Bala, Tanushree

2017-01-01

Highlights: • Ni nanoparticles were synthesized in polymer to form Ni-Polymer composite. • Ni nanoparticles retain their superparamagnetism in the composite. • Ni-Polymer composites showed catalytic activity. - Abstract: Composites comprising of metallic nanoparticles in polymer matrices have allured significant importance due to multifunctionalities. Here a simple protocol has been described to embed Ni nanoparticles in carboxymethyl guar gum (CMGG) polymer. The composite formation helps in the stabilization of Ni nanoparticles which are otherwise prone towards aerial oxidation. Further the nanoparticles retain their superparamagnetic nature and catalytic capacity. Ni-Polymer composite catalyses the reduction of 4-Nitrophenol to 4-Aminophenol very efficiently in presence of NaBH_4, attaining a complete conversion under some experimental conditions. Ni-Polymer composite is well characterized using UV–vis spectroscopy, FTIR, XPS, powder XRD, TGA, SEM and TEM. A detailed magnetic measurement using superconducting quantum interference device-vibrating sample magnetometer (SQUID-VSM) reveals superparamagnetic behaviour of the composite.

Multiple functionalities of Ni nanoparticles embedded in carboxymethyl guar gum polymer: catalytic activity and superparamagnetism

Energy Technology Data Exchange (ETDEWEB)

Sardar, Debasmita [Department of Chemistry, University of Calcutta, 92 A.P.C. Road, Kolkata 700009 (India); Sengupta, Manideepa; Bordoloi, Ankur [Nano Catalysis, Catalytic Conversion and Process Division, CSIR—Indian Institute of Petroleum (IIP), Mohkampur, Dehradun 248005 (India); Ahmed, Md. A.; Neogi, S.K.; Bandyopadhyay, Sudipta [Department of Physics, University of Calcutta, 92 A.P.C. Road, Kolkata 700009 (India); Jain, Ruchi; Gopinath, Chinnakonda S. [Catalysis Division and Center of Excellence on Surface Science, CSIR—National Chemical Laboratory, Dr. Homi Bhabha Road, Pune 411 008 (India); Bala, Tanushree, E-mail: tanushreebala@gmail.com [Department of Chemistry, University of Calcutta, 92 A.P.C. Road, Kolkata 700009 (India)

2017-05-31

Highlights: • Ni nanoparticles were synthesized in polymer to form Ni-Polymer composite. • Ni nanoparticles retain their superparamagnetism in the composite. • Ni-Polymer composites showed catalytic activity. - Abstract: Composites comprising of metallic nanoparticles in polymer matrices have allured significant importance due to multifunctionalities. Here a simple protocol has been described to embed Ni nanoparticles in carboxymethyl guar gum (CMGG) polymer. The composite formation helps in the stabilization of Ni nanoparticles which are otherwise prone towards aerial oxidation. Further the nanoparticles retain their superparamagnetic nature and catalytic capacity. Ni-Polymer composite catalyses the reduction of 4-Nitrophenol to 4-Aminophenol very efficiently in presence of NaBH{sub 4}, attaining a complete conversion under some experimental conditions. Ni-Polymer composite is well characterized using UV–vis spectroscopy, FTIR, XPS, powder XRD, TGA, SEM and TEM. A detailed magnetic measurement using superconducting quantum interference device-vibrating sample magnetometer (SQUID-VSM) reveals superparamagnetic behaviour of the composite.

Structural, kinetic, and docking studies of artificial imine reductases based on biotin-streptavidin technology: an induced lock-and-key hypothesis.

Science.gov (United States)

Robles, Victor Muñoz; Dürrenberger, Marc; Heinisch, Tillmann; Lledós, Agustí; Schirmer, Tilman; Ward, Thomas R; Maréchal, Jean-Didier

2014-11-05

An artificial imine reductase results upon incorporation of a biotinylated Cp*Ir moiety (Cp* = C5Me5(-)) within homotetrameric streptavidin (Sav) (referred to as Cp*Ir(Biot-p-L)Cl] ⊂ Sav). Mutation of S112 reveals a marked effect of the Ir/streptavidin ratio on both the saturation kinetics as well as the enantioselectivity for the production of salsolidine. For [Cp*Ir(Biot-p-L)Cl] ⊂ S112A Sav, both the reaction rate and the selectivity (up to 96% ee (R)-salsolidine, kcat 14-4 min(-1) vs [Ir], KM 65-370 mM) decrease upon fully saturating all biotin binding sites (the ee varying between 96% ee and 45% ee R). In contrast, for [Cp*Ir(Biot-p-L)Cl] ⊂ S112K Sav, both the rate and the selectivity remain nearly constant upon varying the Ir/streptavidin ratio [up to 78% ee (S)-salsolidine, kcat 2.6 min(-1), KM 95 mM]. X-ray analysis complemented with docking studies highlight a marked preference of the S112A and S112K Sav mutants for the SIr and RIr enantiomeric forms of the cofactor, respectively. Combining both docking and saturation kinetic studies led to the formulation of an enantioselection mechanism relying on an "induced lock-and-key" hypothesis: the host protein dictates the configuration of the biotinylated Ir-cofactor which, in turn, by and large determines the enantioselectivity of the imine reductase.

Magnetic susceptibility characterisation of superparamagnetic microspheres

Science.gov (United States)

Grob, David Tim; Wise, Naomi; Oduwole, Olayinka; Sheard, Steve

2018-04-01

The separation of magnetic materials in microsystems using magnetophoresis has increased in popularity. The wide variety and availability of magnetic beads has fuelled this drive. It is important to know the magnetic characteristics of the microspheres in order to accurately use them in separation processes integrated on a lab-on-a-chip device. To investigate the magnetic susceptibility of magnetic microspheres, the magnetic responsiveness of three types of Dynabeads microspheres were tested using two different approaches. The magnetophoretic mobility of individual microspheres is studied using a particle tracking system and the magnetization of each type of Dynabeads microsphere is measured using SQUID relaxometry. The magnetic beads' susceptibility is obtained at four different applied magnetic fields in the range of 38-70 mT for both the mobility and SQUID measurements. The susceptibility values in both approaches show a consistent magnetic field dependence.

Biopolymers coated superparamagnetic Nickel Ferrites: Enhanced biocompatibility and MR imaging probe for breast cancer

Energy Technology Data Exchange (ETDEWEB)

Bano, Shazia, E-mail: shaziaphy@gmail.com [Department of Physics, The Islamia University of Bahawalpur (Pakistan); Zafar, Tayyaba [Department of Physics, The Islamia University of Bahawalpur (Pakistan); Akhtar, Shahnaz [Department of Pharmacy, The Islamia University of Bahawalpur (Pakistan); Buzdar, Saeed Ahmed [Department of Physics, The Islamia University of Bahawalpur (Pakistan); Waraich, Mustansar Mahmood, E-mail: mustansarwaraich@gmail.com [Quaid-e-Azam Medical College B.V. Hospital, Bahawalpur (Pakistan); Afzal, Muhammad [Department of Physics, The Islamia University of Bahawalpur (Pakistan)

2016-11-01

We report evidence for the promising application of bovine serum albumin (BSA), chitosan (CS) or carboxymethyl cellulose (CMC) coated NiFe{sub 2}O{sub 4} cores for improved biocompatibility and enhanced T2 relaxivity, through a single combinatorial approach. Pure nickel-ferrite nano cores (NFs) successfully synthesized by thermolysis, were immobilize with BSA, CS or CMC layer employing a simple cross linking procedure to avoid any significant influence of these biopolymers on the morphology and crystal structure of the cores. Phase, morphology, magnetic hysteresis and surface chemistry were characterized by X-ray diffraction (XRD), Field emission scanning electron microscopy (FE-SEM), vibrating sample magnetometer (VSM) and Fourier transform infrared (FTIR) spectroscopy. The preliminary haemolysis and cell viability experiments show that biopolymers conjugation mitigates the haemolytic effect of the NFs on erythrocytes as the haemolytic index is less than 2% and cell viability is up to 100%, when normalized with the nontreated cells. The relaxivity value of coated NFs is 351±2.6 when compared to 84±0.22 of NFs without biopolymer conjugation. The results demonstrate that BSA, CS or CMC covering on NFs provide a single combinatorial approach to improve the biocompatibility and enhance the relaxivity value. Thus addressing the current challenge of the same with very good contrast for targeting MCF-7 without any further vectorization. - Highlights: • A single combinatorial system for the promising application of biopolymers coated NiFe{sub 2}O{sub 4} cores. • Immobilization of a thin layer of three different biopolymers via a simple approach. • Excellent MR contrast enhancement and targeting of MCF-7 without any further vectorization.

Bead Collage: An Arts-Based Research Method

Science.gov (United States)

Kay, Lisa

2013-01-01

In this paper, "bead collage," an arts-based research method that invites participants to reflect, communicate and construct their experience through the manipulation of beads and found objects is explained. Emphasizing the significance of one's personal biography and experiences as a researcher, I discuss how my background as an…

Elution of Clindamycin and Enrofloxacin From Calcium Sulfate Hemihydrate Beads In Vitro.

Science.gov (United States)

Phillips, Heidi; Boothe, Dawn M; Bennett, R Avery

2015-11-01

To compare the in vitro elution characteristics of clindamycin and enrofloxacin from calcium sulfate hemihydrate beads containing a single antibiotic, both antibiotics, and each antibiotic incubated in the same eluent well. Experimental in vitro study. Calcium sulfate hemihydrate beads were formed by mixing with clindamycin and/or enrofloxacin to create 4 study groups: (1) 160 mg clindamycin/10 beads; (2) 160 mg enrofloxacin/10 beads; (3) 160 mg clindamycin + 160 mg enrofloxacin/10 beads; and (4) 160 mg clindamycin/5 beads and 160 mg enrofloxacin/5 beads. Chains of beads were formed in triplicate and placed in 5 mL phosphate buffered saline (PBS; pH 7.4 and room temperature) with constant agitation. Antibiotic-conditioned PBS was sampled at 14 time points from 1 hour to 30 days. Clindamycin and enrofloxacin concentrations in PBS were determined using high-performance liquid chromatography. Eluent concentrations from clindamycin-impregnated beads failed to remain sufficiently above minimum inhibitory concentration (MIC) for common infecting bacteria over the study period. Enrofloxacin eluent concentrations remained sufficiently above MIC for common wound pathogens of dogs and cats and demonstrated an atypical biphasic release pattern. No significant differences in elution occurred as a result of copolymerization of the antibiotics into a single bead or from individual beads co-eluting in the same eluent well. Clindamycin-impregnated beads cannot be recommended for treatment of infection at the studied doses; however, use of enrofloxacin-impregnated beads may be justified when susceptible bacteria are cultured. © Copyright 2015 by The American College of Veterinary Surgeons.

A NOVEL APPROACH TO SYNTHESIZE CHITOSAN BEADS CROSSLINKED BY EPICHLOROHYDRIN

Institute of Scientific and Technical Information of China (English)

WANG Yongjian; BAI Shu; SUN Yan

2001-01-01

The present investigation describes a novel method for preparing spherical chitosan particles based on crosslinking with epichlorohydrin. Certain amount of pre-crosslinking agent was added to form chitosan gels by traditional inverse phase suspension polymerization. Then the gels were crosslinked by epichlorohydrin at basic condition to obtain chitosan beads. The effects of reaction conditions, such as crosslinking time, the amount of crosslinking agent and the NaOtt concentration,on the physical properties of the chitosan beads were investigated. The beads were found to have more amino groups in the polymer chains than the beads crosslinked by glutaraldehyde. The capacity for copper ions is as high as 40mg/g. The beads have good mechanical strength and can be reused.

A NOVEL APPROACH TO SYNTHESIZE CHITOSAN BEADS CROSSLINKED BY EPICHLOROHYDRIN

Institute of Scientific and Technical Information of China (English)

WANGYongjina; BAIShu; 等

2001-01-01

The present investigation describes a novel method for preparing spherical chitosan particles based on crosslinking with epichlorohydrin.Certain amount of pre-crosslinking agent was added to form chitosan gels by traditional inverse phase suspension polymerization.Then the gels were crosslinked by epichlorohydrin at basic condition to obtain chitosan beads.The effects of reaction conditions,such as crosslinking time,the amount of crosslinking agent and the NaOH concentration,on the physical properties of the chitosan beads were investigated.The beads were found to have more amino groups in the polymer chains than the beads crosslinked by glutaraldehyde.The capacity for copper ions in as high as 40mg/g,The beads have good mechanical strength and can be reused.

Random glycopeptide bead libraries for seromic biomarker discovery

DEFF Research Database (Denmark)

Kracun, Stjepan Kresimir; Cló, Emiliano; Clausen, Henrik

2010-01-01

have developed a random glycopeptide bead library screening platform for detection of autoantibodies and other binding proteins. Libraries were build on biocompatible PEGA beads including a safety-catch C-terminal amide linker (SCAL) that allowed mild cleavage conditions (I(2)/NaBH(4) and TFA...... to other tumor glycoforms by on-bead enzymatic glycosylation reactions with recombinant glycosyltransferases. Hence, we have developed a high-throughput flexible platform for rapid discovery of O-glycopeptide biomarkers and the method has applicability in other types of assays such as lectin...

Fabrication and evaluation of chitosan/NaYF4:Yb3+/Tm3+ upconversion nanoparticles composite beads based on the gelling of Pickering emulsion droplets

International Nuclear Information System (INIS)

Yan, Huiqiong; Chen, Xiuqiong; Shi, Jia; Shi, Zaifeng; Sun, Wei; Lin, Qiang; Wang, Xianghui; Dai, Zihao

2017-01-01

The rare earth ion doped upconversion nanoparticles (UCNPs) synthesized by hydrophobic organic ligands possess poor solubility and low fluorescence quantum yield in aqueous media. To conquer this issue, NaYF 4 :Yb 3+ /Tm 3+ UCNPs, synthesized by a hydrothermal method, were coated with F127 and then assembled with chitosan to fabricate the chitosan/NaYF 4 :Yb 3+ /Tm 3+ composite beads (CS/NaYF 4 :Yb 3+ /Tm 3+ CBs) by Pickering emulsion system. The characterization results revealed that the as-synthesized NaYF 4 :Yb 3+ /Tm 3+ UCNPs with an average size of 20 nm exhibited spherical morphology, high crystallinity and characteristic emission upconversion fluorescence with an overall blue color output. The NaYF 4 :Yb 3+ /Tm 3+ UCNPs were successfully conjugated on the surface of chitosan beads by the gelling of emulsion droplets. The resultant CS/NaYF 4 :Yb 3+ /Tm 3+ CBs showed good upconversion luminescent property, drug-loading capacity, release performance and excellent biocompatibility, exhibiting great potentials in targeted drug delivery and tissue engineering with potential tracking capability and lasting release performance. - Highlights: • NaYF 4 :Yb 3+ /Tm 3+ UCNPs were coated by F127 to improve aqueous dispersibility. • NaYF 4 :Yb 3+ /Tm 3+ UCNPs were assembled with chitosan to fabricate the composite beads (CMs). • Pickering emulsions stabilized by UCNPs exhibited uniform and satisfactory emulsion droplets. • The CMs prepared by the gelling of emulsion droplet preserved upconversion luminescent property. • The resultant CMs showed good drug-loading capacity, release performance and biocompatibility.

Ionically crosslinked alginate–carboxymethyl cellulose beads for the delivery of protein therapeutics

International Nuclear Information System (INIS)

Kim, Min Sup; Park, Sang Jun; Gu, Bon Kang; Kim, Chun-Ho

2012-01-01

Highlights: ► We prepared Fe 3+ crosslinked alginate–carboxymethyl cellulose (AC) beads. ► Different surface and inner morphology of AC beads were observed on volume of CMC. ► AC beads showed minimum swelling degree in acidic condition. ► Protein release from AC beads was to control in gastrointestinal condition. - Abstract: We developed Fe 3+ -crosslinked alginate–carboxymethyl cellulose (AC) beads in various volume ratios by dropping an AC solution into a ferric chloride solution to form protein therapeutic carrier beads. Scanning electron microscopy revealed that the roughness and pore size of the crosslinked beads increased with the volume ratio of the carboxymethyl cellulose. Fourier transform-infrared analysis revealed the formation of a three-dimensional bonding structure between the anionic polymeric chains of AC and the Fe 3+ ions. The degree of swelling and the release profile of albumin from the beads were investigated under simulated gastrointestinal conditions (pH 1.2, 4.5, and 7.4). The Fe 3+ -crosslinked AC beads displayed different degrees of swelling and albumin release for the various AC volume ratios and under various pH conditions. An in vitro release test was used to monitor the controlled release of albumin from the AC beads under simulated gastrointestinal conditions over 24 h. The Fe 3+ -crosslinked AC beads protected and controlled the release of protein, demonstrating that such beads present a promising protein therapeutic carrier for the oral delivery.

A magnetic bead-based method for concentrating DNA from human urine for downstream detection.

Science.gov (United States)

Bordelon, Hali; Russ, Patricia K; Wright, David W; Haselton, Frederick R

2013-01-01

Due to the presence of PCR inhibitors, PCR cannot be used directly on most clinical samples, including human urine, without pre-treatment. A magnetic bead-based strategy is one potential method to collect biomarkers from urine samples and separate the biomarkers from PCR inhibitors. In this report, a 1 mL urine sample was mixed within the bulb of a transfer pipette containing lyophilized nucleic acid-silica adsorption buffer and silica-coated magnetic beads. After mixing, the sample was transferred from the pipette bulb to a small diameter tube, and captured biomarkers were concentrated using magnetic entrainment of beads through pre-arrayed wash solutions separated by small air gaps. Feasibility was tested using synthetic segments of the 140 bp tuberculosis IS6110 DNA sequence spiked into pooled human urine samples. DNA recovery was evaluated by qPCR. Despite the presence of spiked DNA, no DNA was detectable in unextracted urine samples, presumably due to the presence of PCR inhibitors. However, following extraction with the magnetic bead-based method, we found that ∼50% of spiked TB DNA was recovered from human urine containing roughly 5×10(3) to 5×10(8) copies of IS6110 DNA. In addition, the DNA was concentrated approximately ten-fold into water. The final concentration of DNA in the eluate was 5×10(6), 14×10(6), and 8×10(6) copies/µL for 1, 3, and 5 mL urine samples, respectively. Lyophilized and freshly prepared reagents within the transfer pipette produced similar results, suggesting that long-term storage without refrigeration is possible. DNA recovery increased with the length of the spiked DNA segments from 10±0.9% for a 75 bp DNA sequence to 42±4% for a 100 bp segment and 58±9% for a 140 bp segment. The estimated LOD was 77 copies of DNA/µL of urine. The strategy presented here provides a simple means to achieve high nucleic acid recovery from easily obtained urine samples, which does not contain inhibitors of PCR.

exploring traditional glass bead making techniques in jewellery

African Journals Online (AJOL)

User

Glass bead making techniques and their mass production will help the individual ... communicate cultural values in a symbolic lan- guage which ..... Surface of most of the new beads were rough ... tourism potential to be developed further to.

Chinese and Venetian glass beads excavated from Fais Island in Micronesia

International Nuclear Information System (INIS)

Intoh, Michiko

1997-01-01

Full text: Over 830 glass beads were excavated from a late prehistoric cemetery site on Fais Island in the Caroline Islands, Micronesia In one of the 13 excavated burials a young woman had more that 310 glass beads around her wrist. Bone collagen from this burial was dated by AMS to 387 + 64 BP. The associated glass beads were classified into three groups based on colour and size. A sample from each group was examined for evidence of manufacturing technique. The chemical composition was determined using an X-ray microanalyser. The first group consisted of more than 300 pale green, transparent glass beads which are less than 2 mm in diameter. The chemical composition is high in PbO (75.22%) while low in MgO. Such a high lead content is characteristic of Chinese glass. The manufacturing technique could not be determined because the surfaces were too eroded. The second group contains several yellow, translucent glass beads. The chemical composition is also high in PbO (54.8%) and low in MgO. The beads were made by winding. The combination of winding and high lead strongly indicates that the beads were made in China. The third group had only one white, translucent glass bead. It has particular white stripes which suggest that it is a 'gooseberry' bead which was made in Venice between the sixteenth and eighteenth centuries. In conclusion, both Chinese and Venetian glass beads co-existed on Fais Island around the time of European contact. They are likely to have been brought in from an area which had access to both beads. Island South-East Asia is tentatively considered to be the source area

Cooling Rates of Lunar Volcanic Glass Beads

Science.gov (United States)

Hui, Hejiu; Hess, Kai-Uwe; Zhang, Youxue; Peslier, Anne; Lange, Rebecca; Dingwell, Donald; Neal, Clive

2016-01-01

It is widely accepted that the Apollo 15 green and Apollo 17 orange glass beads are of volcanic origin. The diffusion profiles of volatiles in these glass beads are believed to be due to degassing during eruption (Saal et al., 2008). The degree of degassing depends on the initial temperature and cooling rate. Therefore, the estimations of volatiles in parental magmas of lunar pyroclastic deposits depend on melt cooling rates. Furthermore, lunar glass beads may have cooled in volcanic environments on the moon. Therefore, the cooling rates may be used to assess the atmospheric condition in an early moon, when volcanic activities were common. The cooling rates of glasses can be inferred from direct heat capacity measurements on the glasses themselves (Wilding et al., 1995, 1996a,b). This method does not require knowledge of glass cooling environments and has been applied to calculate the cooling rates of natural silicate glasses formed in different terrestrial environments. We have carried out heat capacity measurements on hand-picked lunar glass beads using a Netzsch DSC 404C Pegasus differential scanning calorimeter at University of Munich. Our preliminary results suggest that the cooling rate of Apollo 17 orange glass beads may be 12 K/min, based on the correlation between temperature of the heat capacity curve peak in the glass transition range and glass cooling rate. The results imply that the parental magmas of lunar pyroclastic deposits may have contained more water initially than the early estimations (Saal et al., 2008), which used higher cooling rates, 60-180 K/min in the modeling. Furthermore, lunar volcanic glass beads could have been cooled in a hot gaseous medium released from volcanic eruptions, not during free flight. Therefore, our results may shed light on atmospheric condition in an early moon.

Mössbauer studies of superparamagnetic ferrite nanoparticles for functional application

International Nuclear Information System (INIS)

Mažeika, K.; Jagminas, A.; Kurtinaitienė, M.

2013-01-01

Nanoparticles of CoFe 2 O 4 and MnFe 2 O 4 prepared for functional applications in nanomedicine were studied using Mössbauer spectrometry. Superparamagnetic properties of nanoparticles of different size and composition were compared applying collective excitations and multilevel models for the description of the Mössbauer spectra.

Transarterial chemoembolization with drug-eluting beads in hepatocellular carcinoma

Science.gov (United States)

Nam, Hee Chul; Jang, Bohyun; Song, Myeong Jun

2016-01-01

Transarterial chemoembolization (TACE) is a widely used standard treatment for patients with hepatocellular carcinoma (HCC) who are not suitable candidates for curative treatments. The rationale for TACE is that intra-arterial chemotherapy using lipiodol and chemotherapeutic agents, followed by selective vascular embolization, results in a strong cytotoxic effect as well as ischemia (conventional TACE). Recently, drug-eluting beads (DC Beads®) have been developed for transcatheter treatment of HCC to deliver higher doses of the chemotherapeutic agent and to prolong contact time with the tumor. DC Beads® can actively sequester doxorubicin hydrochloride from solution and release it in a controlled sustained fashion. Treatment with DC Beads® substantially reduced the amount of chemotherapeutic agent that reached the systemic circulation compared with conventional, lipiodol-based regimens, significantly reducing drug-related adverse events. In this article, we describe the treatment response, survival, and safety of TACE used with drug-eluting beads for the treatment of HCC and discuss future therapeutic possibilities. PMID:27833376

Bead Capture and Release on a Magnetic Sensor in a Microfluidic System

DEFF Research Database (Denmark)

Dalslet, Bjarke Thomas; Damsgaard, Christian Danvad; Freitas, S.C.

Planar Hall effect magnetic sensors for detection of biological agents using surface treated magnetic beads are integrated with a fluid injection system. The response of the sensors is used to evaluate bead capture rates for different bead concentrations c and fluid flow rates Q, and to monitor...... subsequent removal of the beads. It is found that the capture rate scales directly with c and that it depends linearly on Q. At low Q the capture rate is only partially due to gravitational sedimentation of beads. At higher Q an additional term proportional to Q becomes important, which is attributed...... to capture of beads by the magnetic fields near the sensor. It is shown that beads can be washed off the sensor surface....

Analysis of pirlimycin residues in beef muscle, milk and honey by a biotin-streptavidin-amplified enzyme-linked immunosorbent assay

Science.gov (United States)

Food contamination caused by veterinary drug residues is a world-wide public health concern and requires continuous monitoring. In this paper, we describe a biotin–streptavidin-amplified ELISA (BA-ELISA) for detecting pirlimycin residues in beef, milk, and honey. The IC50 value of the BA-ELISA was...

Synthesis and super-paramagnetic properties of neodymium ferrites nanorods

Energy Technology Data Exchange (ETDEWEB)

El moussaoui, H. [Institute of Nanomaterials and Nanotechnologies, MAScIR, Rabat (Morocco); Laboratoire of Magnetism and the Physics of the High Energies, URAC 12, Departement of Physique, Faculty of Science, Mohammed V- Agdal University, BP 1014, Rabat (Morocco); Mounkachi, O., E-mail: o.mounkachi@mascir.com [Institute of Nanomaterials and Nanotechnologies, MAScIR, Rabat (Morocco); Masrour, R. [Laboratory of Materials, Processes, Environment and Quality, Cady Ayyed University, National School of Applied Sciences, Route Sidi Bouzid, BP 63, 46000 Safi (Morocco); Hamedoun, M., E-mail: hamedoun@hotmail.com [Institute of Nanomaterials and Nanotechnologies, MAScIR, Rabat (Morocco); Hlil, E.K. [Institut Néel, CNRS-UJF, B.P. 166, 38042 Grenoble Cedex (France); Benyoussef, A. [Institute of Nanomaterials and Nanotechnologies, MAScIR, Rabat (Morocco); Laboratoire of Magnetism and the Physics of the High Energies, URAC 12, Departement of Physique, Faculty of Science, Mohammed V- Agdal University, BP 1014, Rabat (Morocco); Hassan II Academy of Science and Technology, Rabat (Morocco)

2013-12-25

Highlights: •Magnetic properties of Neodymium nanorods depend on calcination temperature. •The as-synthesized Nd ferrite nanorods are superparamagnetic at room temperature. •The blocking temperature is higher than room temperature. -- Abstract: In this work we report the microstructural characterization and the magnetic properties of neodymium ferrites (NdFe{sub 2}O{sub 4}) nanorods prepared by well controlled co-precipitation method. The effect of annealing temperature on the structure, morphology and magnetic properties of NdFe{sub 2}O{sub 4} has been investigated. The transmission electron microscopy (TEM) observations revealed that the as-prepared nanoparticles have rods-like shape with the average diameter ranging from 5 to 14 nm and uniform length. The magnetic measurements show that the as-synthesized nanorods have a superparamagnetic behavior at room temperature, with a blocking temperature of 360 K and magnetic anisotropy constant of 2.8 × 10{sup 5} ergs/cm{sup 3}. The magnetization and coercitivity at room temperature are increased from 26 to 34 emu/g and from 151 to 171 Oe with increasing annealing temperature from 400 to 600 °C, respectively.

Release Behavior and Toxicity Profiles towards Leukemia (WEHI-3B) Cell Lines of 6-Mercaptopurine-PEG-Coated Magnetite Nanoparticles Delivery System

OpenAIRE

Dorniani, Dena; Kura, Aminu Umar; Hussein-Al-Ali, Samer Hasan; Hussein, Mohd Zobir bin; Fakurazi, Sharida; Shaari, Abdul Halim; Ahmad, Zalinah

2014-01-01

The coating of an active drug, 6-mercaptopurine, into the iron oxide nanoparticles-polyethylene glycol (FNPs-PEG) in order to form a new nanocomposite, FPEGMP-2, was accomplished using coprecipitation technique. The resulting nanosized with a narrow size distribution magnetic polymeric particles show the superparamagnetic properties with 38.6 emu/g saturation magnetization at room temperature. Fourier transform infrared spectroscopy and the thermal analysis study supported the formation of th...

Polymerized serum albumin beads for use as slow-release adjuvants

International Nuclear Information System (INIS)

Martin, M.E.D.

1987-02-01

Experimental vaccines have been made by covalently bonding virus particles into polymerized rabbit serum albumin beads. Using Nodamura virus as a model antigen, these model vaccines induced specific humoral antibody production, comparable with that achieved using Freund's adjuvants. Virus specific antibodies were also induced when Nodamura virus was covalently attached to the bead surface using different crosslinkers. However, when poliovirus type 2 (Sabin strain) was polymerized into beads, the levels of neutralizing antibodies were insignificant compared with control aqueous vaccines. The synthetic immunostimulator, muramyl dipeptide, was included with bead vaccines in an attempt to potentiate the immune response. Immunostimulation is achieved by a slow release of antigen coinciding with the gradual breakdown of bead structure. Methods used include radio-iodination and radioimmunoassay. 65 figs., 6 tabs., 173 refs