Determination of concentration and binding affinity of antibody fragments by use of surface plasmon resonance

An assay method using a surface plasmon resonance (SPR) biosensor has been developed that allows quantitative measurement of the specific antibody concentration in crude materials. By injecting non-labeled antibody samples onto a biosensor surface on which antigen was immobilized at high densities, the concentration of active antibodies can be accurately measured. To clarify applicability of this method to pharmacokinetic studies, the concentration of active antibodies in mouse plasma was measured for 4 h after injection of antibodies in mice. Although this period of measurement might be insufficient for determining the pharmacokinetics of blood pool clearance, this method has some advantages over conventional methods in measurement of single-chain antibody fragment (scFv) concentrations. Using the SPR biosensor, scFv and antibodies without epitope tag peptides were easily detected in real time, requiring as little as 20 mul of blood sample. Moreover, from the apparent dissociation rate in the dissociation phase of the sensorgrams, we could identify whether the antibody fragments existed as bivalent or monovalent in animal blood. We also evaluated the antigen binding activity of the scFvs against human CD47 and found scFvs had slightly weak affinity to their antigen (K(D), about 10 nM) compared with F(ab')2 and Fab' fragments (K(D), about 3-4 nM). This assay method promises to be a convenient tool for quality control, screening, and simple pharmacokinetic analysis of antibody fragments and other recombinant proteins not having epitope tags.     

Depletion of glutathione as a cause of the promotive effects of polygodial, a sesquiterpene on the production of reactive oxygen species in Saccharomyces cerevisiae

The pungent sesquiterpenoid unsaturated dialdehyde, polygodial, exhibited a strong yeastcidal activity against the cells of Saccharomyces cerevisiae, in which production of reactive oxygen species (ROS) at a significant level could be detected with a fluorescent probe. The production of ROS in polygodial-treated cells was further confirmed by its elimination and the accompanying protection against yeastcidal effects in the presence of antioxidants such as L-ascorbate and alpha-tocopherol (alpha-TOH). Polygodial could accelerate ROS production only in cells of the wild-type grande strain but not in those of the respiratory-deficient petite mutant (rho0), indicating the role of the mitochondrial electron transport chain in the production of ROS. Unlike the case with antimycin A which accelerates ROS production by directly targeting the mitochondrial electron flow, polygodial caused depletion of cytoplasmic and mitochondrial glutathione which functions in estiminating ROS inevitably generated during aerobic growth. Polygodial-mediated depletion of intracellular glutathione was possibly dependent on a direct interaction between its enal moiety and the sulfhydryl group of the cysteine in glutathione by a Michael-type reaction.     

Microwave frequency effect in the formation of Au nanocolloids in polar and non-polar solvents

Given earlier observations that microwave frequencies can have a substantial effect on the photoactivity of a well-known photocatalyst (TiO(2)), in the synthesis of 3,6-diphenyl-4-n-butylpyridazine through a Diels-Alder process, and in the one-pot solvent-free synthesis of a room-temperature ionic liquid, we proceeded to examine the frequency effects of the 5.8 and 2.45 GHz microwave (MW) radiation in the synthesis of gold nanoparticles in non-polar media, such as oleylamine, which have a low dielectric constant (ε'), and we further examine differences in shape and size under otherwise identical temperature conditions when the synthesis of the gold nanoparticles was carried out in an ethylene glycol polar medium in the presence of polyvinylpyrrolidone. Whereas a change in microwave frequency from 2.45 to 5.8 GHz at equal microwave power levels led to the synthesis of gold nanoparticles in the non-polar media, a change in the microwave frequency had no effect on the size and shape of the gold nanoparticles synthesized in polar media for identical microwave power levels.     

Multiscale modeling of calcium dynamics in ventricular myocytes with realistic transverse tubules

Spatial-temporal Ca(2+) dynamics due to Ca(2+) release, buffering, and reuptaking plays a central role in studying excitation-contraction (E-C) coupling in both normal and diseased cardiac myocytes. In this paper, we employ two numerical methods, namely, the meshless method and the finite element method, to model such Ca(2+) behaviors by solving a nonlinear system of reaction-diffusion partial differential equations at two scales. In particular, a subcellular model containing several realistic transverse tubules (or t-tubules) is investigated and assumed to reside at different locations relative to the cell membrane. To this end, the Ca(2+) concentration calculated from the whole-cell modeling is adopted as part of the boundary constraint in the subcellular model. The preliminary simulations show that Ca(2+) concentration changes in ventricular myocytes are mainly influenced by calcium release from t-tubules.     

Reflectance and Shape Estimation with a Light Field Camera Under Natural Illumination

International Journal of Computer Vision - Reflectance and shape are two important components in visually perceiving the real world. Inferring the reflectance and shape of an object through cameras...     

Position Dependent Response of Superconducting MgB2 Neutron Detectors Studied by Pulsed Laser Irradiation

The MgB₂ detector consists of 200?nm thick MgB₂ thin film meander structure of 3?μm line width protected by 300?nm thick SiO layer. The electrical connections were provided by 150?nm thick Nb. The devices were placed in a 4?K refrigerator to investigate the behavior at temperatures below T _c. The position-dependent response of the MgB₂ detector can be investigated by scanning a laser spot with the aid of the XYZ piezo-driven stage and an optical fiber with a focusing lens. We found that the inhomogeneity in a detector is very critical to specify the detection efficiency since the detector has the sensitivity only in a narrow temperature regime near T _c.     

Experimental and numerical investigation on compressor cascade flows with tip clearance at a low Reynolds number condition

High flow rate aeroengines typically employ axial flow compressors,where aerodynamic loss is predominantly due to secondary flow features such as tip leakage and corner vortices.In very high altitude missions,turbomachinery operates at low density ambient atmosphere,and the recent trend toward more compact engine core inevitably leads to the reduction of blade size,which in turn increases the relative height of the blade tip clearance.Low Reynolds number flowfield as a result of these two factors amplifies the relative importance of secondary flow effects.This paper focuses on the behavior of tip leakage flow,investigating by use of both experimental and numerical approaches.In order to understand the complex secondary flow behavior,cascade tests are usually conducted using intrusive probes to determine the loss.However relatively few experimental studies are published on tip leakage flows which take into account the interaction between a rotating blade row and its casing wall.Hence a new linear cascade facility has been designed with a moving belt casing in order to reproduce more realistic flowfield as encountered by a rotating compressor row.Numerical simulations were also performed to aid in the understanding of the complex flow features.The experimental results indicate a significant difference in the flowfield when the moving belt casing is present.The numerical simulations reveal that the leakage vortex is pulled by the shearing motion of the endwall toward the pressure side of the adjacent blade.The results highlight the importance of casing wall relative motion in analyzing leakage flow effects.