Efficient DNA ligation by selective heating of DNA ligase with a radio frequency alternating magnetic field.

We present a simple method for efficient DNA ligation utilizing the heat generation of ferromagnetic particles subjected to an ac magnetic field. We carry out the ligation of DNA fragments with cohesive ends using T4 DNA ligase immobilized on the surface of ferromagnetic particles. When a radio frequency alternating magnetic field is applied, ferromagnetic particles dissipate heat and DNA ligase on the particles is selectively heated up and activated with little influence on the annealing of DNA ends, as a result of which the ligation efficiency increases. We show that the ligation efficiency increases with an increase in the field amplitude.

Encouragement of Enzyme Reaction Utilizing Heat Generation from Ferromagnetic Particles Subjected to an AC Magnetic Field.

We propose a method of activating an enzyme utilizing heat generation from ferromagnetic particles under an ac magnetic field. We immobilize α-amylase on the surface of ferromagnetic particles and analyze its activity. We find that when α-amylase/ferromagnetic particle hybrids, that is, ferromagnetic particles, on which α-amylase molecules are immobilized, are subjected to an ac magnetic field, the particles generate heat and as a result, α-amylase on the particles is heated up and activated. We next prepare a solution, in which α-amylase/ferromagnetic particle hybrids and free, nonimmobilized chitinase are dispersed, and analyze their activities. We find that when the solution is subjected to an ac magnetic field, the activity of α-amylase immobilized on the particles increases, whereas that of free chitinase hardly changes; in other words, only α-amylase immobilized on the particles is selectively activated due to heat generation from the particles.

Activity of lipase and chitinase immobilized on superparamagnetic particles in a rotational magnetic field.

We immobilize hydrolases such as lipase and chitinase on superparamagnetic particles, which are subjected to a rotational magnetic field, and measure the activities of the enzymes. We find that the activities of lipase and chitinase increase in the rotational magnetic field compared to those in the absence of a magnetic field and reach maximum at certain frequencies. The present methodology may well be utilized for the design and development of efficient micro reactors and micro total analysis systems (µ-TASs).

Curcumin loaded-PLGA nanoparticles conjugated with Tet-1 peptide for potential use in Alzheimer's disease.

Alzheimer's disease is a growing concern in the modern world. As the currently available medications are not very promising, there is an increased need for the fabrication of newer drugs. Curcumin is a plant derived compound which has potential activities beneficial for the treatment of Alzheimer's disease. Anti-amyloid activity and anti-oxidant activity of curcumin is highly beneficial for the treatment of Alzheimer's disease. The insolubility of curcumin in water restricts its use to a great extend, which can be overcome by the synthesis of curcumin nanoparticles. In our work, we have successfully synthesized water-soluble PLGA coated- curcumin nanoparticles and characterized it using different techniques. As drug targeting to diseases of cerebral origin are difficult due to the stringency of blood-brain barrier, we have coupled the nanoparticle with Tet-1 peptide, which has the affinity to neurons and possess retrograde transportation properties. Our results suggest that curcumin encapsulated-PLGA nanoparticles are able to destroy amyloid aggregates, exhibit anti-oxidative property and are non-cytotoxic. The encapsulation of the curcumin in PLGA does not destroy its inherent properties and so, the PLGA-curcumin nanoparticles can be used as a drug with multiple functions in treating Alzheimer's disease proving it to be a potential therapeutic tool against this dreaded disease.

Cluster-cluster aggregations of superparamagnetic particles in a rotational magnetic field.

We investigate the cluster-cluster aggregations of superparamagnetic particles in a rotational magnetic field numerically by the Brownian dynamics method, focusing on the cases of ϕ = 0.01 and 0.03 and Ma = 0, 0.001, 0.01, and 0.1, where ϕ is the area fraction of superparamagnetic particles and Ma is the Mason number, i.e., the ratio of viscous drag to magnetic force acting on a magnetic particle. We clarify the effect of ϕ and Ma on the cluster-cluster aggregation process from the point of view of dynamic scaling law.

Ordered complex structures formed by paramagnetic particles via self-assembly under an ac/dc combined magnetic field.

We apply ac and dc magnetic fields simultaneously in orthogonal directions to each other to a solution, in which paramagnetic microparticles are dispersed, and show that complex secondary structures composed of oscillating chain clusters, that is, long linear clusters interconnected by T-, L-, and criss-cross-junctions, are self-assembled. Disklike clusters are formed at some junctions and the number of disklike clusters increases as the frequency of the ac magnetic field increases. We finally show that the angle between long linear clusters can be altered by changing the ratio of the intensities of the ac and dc magnetic fields.

Aptamer conjugated magnetic nanoparticles as nanosurgeons.

Magnetic nanoparticles have shown promise in the fields of targeted drug delivery, hyperthermia and magnetic resonance imaging (MRI) in cancer therapy. The ability of magnetic nanoparticles to undergo surface modification and the effect of external magnetic field in the dynamics of their movement make them an excellent nanoplatform for cancer destruction. Surgical removal of cancerous or unwanted cells selectively from the interior of an organ or tissue without any collateral damage is a serious problem due to the highly infiltrative nature of cancer. To address this problem in surgery, we have developed a nanosurgeon for the selective removal of target cells using aptamer conjugated magnetic nanoparticles controlled by an externally applied three-dimensional rotational magnetic field. With the help of the nanosurgeon, we were able to perform surgical actions on target cells in in vitro studies. LDH and intracellular calcium release assay confirmed the death of cancer cells due to the action of the nanosurgeon which in turn nullifies the possibility of proliferation by the removed cells. The nanosurgeon will be a useful tool in the medical field for selective surgery and cell manipulation studies. Additionally, this system could be upgraded for the selective removal of complex cancers from diverse tissues by incorporating various target specific ligands on magnetic nanoparticles.

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

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).

Label-free determination of the number of biomolecules attached to cells by measurement of the cell's electrophoretic mobility in a microchannel.

We developed a label-free method for a determination of the number of biomolecules attached to individual cells by measuring the electrophoretic mobility of the cells in a microchannel. The surface of a biological cell, which is dispersed in aqueous solution, is normally electrically charged and the charge quantity at the cell's surface is slightly changed once antibody molecules are attached to the cell, based on which we detect the attachment of antibody molecules to the surface of individual red blood cells by electrophoretic mobility measurement. We also analyzed the number of antibody molecules attached to the cell's surface using a flow cytometer. We found that there is a clear correlation between the number of antibody molecules attached to the individual cells and the electrophoretic mobility of the cells. The present technique may well be utilized not only in the field of cell biology but also in the medical and pharmaceutical industries.

Ring-chain structural transitions in a ferromagnetic particles system induced by a dc magnetic field.

We carry out statistical analysis of cluster structures formed by ferromagnetic particles in a dc magnetic field based on a flexible chain model. In the absence of a magnetic field, ferromagnetic particles tend to form ring structures. When a dc magnetic field is applied, the rings are broken into chains if the intensity of the field is sufficiently high since the dipole moment of each particle is aligned in the field direction. We analyze the conditions under which ring-chain structural transitions occur and clarify the dependence of the probabilities of the creation of ring and chain structures on control parameters such as the strength of the dipole-dipole interactive energy between particles, the intensity of the magnetic field and the cluster size.

Tumbling motion of magnetic particles on a magnetic substrate induced by a rotational magnetic field.

We analyze the dynamics of paramagnetic particles on a paramagnetic substrate under a rotational magnetic field. When the paramagnetic particles are subjected to a rotational magnetic field, the rotational plane of which is perpendicular to the substrate surface, the particles form chain clusters caused by the dipole-dipole interaction between the particles and these clusters display a tumbling motion under certain conditions. In this case, the angular momentum of the clusters is converted to a translational one through the force of friction acting between the particles and substrate and, as a result, the clusters move along the surface of the substrate. We analyze the conditions under which the tumbling motion occurs and the dependence of the translational velocity of a cluster on the control parameters by the Stokesian dynamics method. Based on the dynamics of magnetic particles, we propose a method of manipulating nano- and microparticles using a rotational magnetic field. We demonstrate the manipulation of magnetic and nonmagnetic particles, a carbon nanotube, and a biological cell.

Magnetic carbon nanotubes: synthesis by electrostatic self-assembly approach and application in biomanipulations.

Magnetic multiwalled carbon nanotubes (MWNTs) were facilely prepared by the electrostatic self-assembly approach. Poly(2-diethylaminoethyl methacrylate) (PDEAEMA) was covalently grafted onto the surfaces of MWNTs by MWNT-initiated in situ atom transfer radical polymerization (ATRP) of 2-diethylaminoethyl methacrylate (DEAEMA). The PDEAEMA-grafted MWNTs were quaternized with methyl iodide (CH(3)I), resulting in cationic polyelectrolyte-grafted MWNTs (MWNT-PAmI). Magnetic iron oxide (Fe(3)O(4)) nanoparticles were loaded onto the MWNT surfaces by electrostatic self-assembling between MWNT-PAmI and Fe(3)O(4), affording magnetic nanotubes. The assembled capability of the nanoparticles can be adjusted to some extent by changing the feed ratio of Fe(3)O(4) to MWNT-PAmI. The obtained magnetic nanotubes were characterized with TEM, EDS, STEM, and element mapping analyses. TEM and EDS measurements confirmed the nanostructures and the components of the resulting nanoobjects. The magnetic nanotubes were assembled onto sheep red blood cells in a phosphate buffer solution, forming magnetic cells. The blood cells attached with or without magnetic nanotubes can be selectively manipulated in a magnetic field. These results promise a general and efficient strategy to magnetic nanotubes and the fascinating potential of such magnetic nanoobjects in applications of bionanoscience and technology.

Dynamics of disklike clusters formed in a magnetorheological fluid under a rotational magnetic field.

We investigate the cluster formations and dynamics in a magnetorheological fluid under a rotational magnetic field focusing on the case of a relatively high volume fraction. We find that isotropic disklike clusters, which rotate more slowly than the field rotation, are formed at low Mason numbers (the ratio of viscous to magnetic forces) and, what is more, we show short rod clusters, which rotate stably thanks to the low Mason numbers and circulate along the surface of the disklike clusters. The circulation velocity of the surface particles is much higher than the rotational surface velocity of the rigid disklike clusters.

Statistical analysis of two-dimensional cluster structures composed of ferromagnetic particles based on a flexible chain model.

We investigate two-dimensional cluster structures composed of ferromagnetic colloidal particles, based on a flexible chain model, by the configurational-bias Monte Carlo method. We clarify the dependence of the probabilities of the creation of different types of clusters on the dipole-dipole interactive energy and the cluster size.

Nonequilibrium Brownian dynamics analysis of negative viscosity induced in a magnetic fluid subjected to both ac magnetic and shear flow fields.

We study the rheological and magnetic characteristics of a magnetic fluid. The system, which we investigate, is as follows. Ferromagnetic particles are dispersed in a solvent, which is subjected to both ac magnetic and shear flow fields. The translational and rotational motions of particles are calculated by the Brownian dynamics method based on Langevin equations and the rheological and magnetic characteristics of the magnetic fluid system are estimated. First, we investigate the rheological and magnetic characteristics of the system in a dc magnetic field and then we analyze the effect of an ac magnetic field on those characteristics. We find that the negative viscosity effect is induced at a certain frequency range of the ac magnetic field. We also find that there are two main mechanisms responsible for the occurrence of negative viscosity. (1) Resonance between the rotational motions of the dipoles of particles and the fluctuation of ac magnetic fields occurs when applied magnetic fields are weak compared to the shear rate, in which case particles can still rotate in magnetic fields. Beyond this resonance frequency, negative viscosity appears. (2) The magnetic dipole moments of particles are forced to stay in the direction of the magnetic field when strong magnetic fields are applied in relatively low shear flow fields. However, negative viscosity occurs when the frequency of external magnetic fields exceeds a critical value, in which case the dipoles rotate continuously in a shear flow without stopping. In both cases, the mean angular velocity of the particles becomes higher than that of the solvent.