Development of a High-Affinity Antibody-Binding Peptide for Site-Specific Modification

Immunoglobulin G (IgG)-binding peptides such as 15-IgBP are convenient tools for the site-specific modification of antibodies and the preparation of homogeneous antibody–drug conjugates. A peptide such as 15-IgBP can be selectively crosslinked to the fragment crystallizable region of human IgG in an affinity-dependent manner via the ϵ-amino group of Lys8. Previously, we found that the peptide 15-Lys8Leu has a high affinity (K_d=8.19 nM) due to the presence of the γ-dimethyl group in Leu8. The primary amino group required for the crosslinking to the antibodies has, however, been lost. Here, we report the design and synthesis of a novel unnatural amino acid, 4-(2-aminoethylcarbamoyl)leucine (Aecl), which possesses both the γ-dimethyl fragment and a primary amino group. A peptide containing Aecl8 (15-Lys8Aecl) was synthesized and showed a binding affinity ten times higher (K_d=24.3 nM) than that of 15-IgBP (K_d=267 nM). Fluorescein isothiocyanate (FITC)-labeled 15-Lys8Aecl with an N-hydroxy succinimide ester at the side chain of Aecl8 (FITC-15-Lys8Aecl(OSu)) successfully labeled an antibody (trastuzumab, Herceptin^®) with the fluorophore. This peptide scaffold has both strong binding affinity and crosslinking capability, and could be a useful tool for the selective chemical modification of antibodies with molecules of interest such as drugs.     

Residues of doxycycline and oxytetracycline in eggs after medication via drinking water to laying hens

Doxycycline (DOTC) and oxytetracycline (OTC) were dissolved in drinking water (0.5 g/l) and supplied to laying hens for 7 consecutive days. Eggs laid were collected daily during and after medication, and the antibiotic concentrations in the yolk and albumin were determined by the cup-plate method with Bacillus cereus var. mycoides ATCC 11778. The concentrations of both antibiotics were increased in yolk day by day with the advance in medication, reached peaks 2 days after withdrawal and then declined gradually. Mean peak concentrations in the yolk were 6.70 micrograms/g for DOTC and 1.42 micrograms/g for OTC. Peak concentrations in the albumin occurred in the middle stage of medication, where the mean values were 12.24 micrograms/g for DOTC and 1.03 micrograms/g for OTC. DOTC was detected in albumin until 24 days after withdrawal and for 2 days more in yolk than in albumin. OTC was detected in yolk until 9 days after withdrawal. The depletion period of OTC was shorter for the albumin, where the residue disappeared in all eggs 6 days after withdrawal. In spite of similarities between DOTC and OTC in structure, DOTC was deposited in higher concentrations and lasted for a longer period in eggs. This characteristic was considered due to its greater lipophilicity, closely correlated with oral absorption and tissue penetration.     

Preparation and Optical Nonlinear Property of Sol–Gel-Derived Cu─SiO2 Thin Films

Cu-metal-doped glass films having a Cu:Si atomic ratio of 0.05 ± 0.002 were successfully prepared by a sol-gel method using a dipping technique. The appearance of surface plasmon of Cu metal at about 570 nm was observed after heat treatment at or above 700°C. The third-order nonlinear susceptibility (x³) was as high as 5.0 × 10^–8 esu at 570 nm.     

Fullerenes in chinese Ink. A Correction

Re-determination of fullerenes in Chinese ink sticks and their raw soots by reverse-phase HPLC analysis revealed much smaller contents of C₆₀ and C₇₀ than previously reported, 3 ppm at most. The older sticks tend to yield less C₆₀ than the more recent ones.     

Effect of the Manganese Valence State on the Electrical Conductivity of Barium Titanate

The effect of defect structure on electrical conductivity for Mn-doped BaTiO₃ at high temperatures was studied experimentally and theoretically. The measured conductivity was proportional to P_o2^−1/4 in the n -type region, and to P _o2^+1/6 in the p -type region in the temperature range between 900° and 1000°C. The results agree with calculations on the basis of the equilibrium equation for the valence change of Mn ions between tetravalent and trivalent. In the p -type region, the concentration of trivalent Mn ions decreases with P _o2^−1/6.     

Antioxidant activity of pea bean (Phaseolus vulgaris L.) extract

Antioxidative activity of pea bean (Phaseolus vulgaris L.) extract was evaluated by using a linoleic acid system, and the methanol extract exhibited strong antioxidative activity as measured by the thiocyanate method. The crude methanol extract was partitioned between then-butanol phase (BP) and the water phase (WP). Then, the antioxidative activity of the BP and the WP was determined by using a linoleic acid system. The WP showed strong antioxidative activity, while BP showed only weak activity as measured by the thiocyanate method. Next, the synergistic antioxidative action of WP with α-tocopherol was examined by using linoleic acid and liposome systems. The WP had a synergistic effect with α-tocopherol in both the food model and liposome systems. For purification and isolation of the antioxidative substances of the pea bean, preparative high-performance liquid chromatography was carried out with an octadecylsilyl column. Five fractions were collected, and antioxidative activity was determined in a linoleic acid system. Although fraction 1 had strong activity by the thiocyanate method, the purification of this active fraction was difficult; therefore, the partly characterized active fraction was investigated. The contents of total phenolics and sugars were 0.31±0.01 mg/g of fraction 1 and 406.1±0.1 mg/g, respectively. The ninhydrin chromogenic reaction was positive, and the ultraviolet absorption spectral λ max value in distilled water was 264.0 nm, indicating that the water-soluble antioxidative components from pea bean may be a new type of antioxidant. Isolation and identification are currently being investigated.     

Study of the degradation of polyurethanes. I. The effect of various metal acetylacetonates on the photodegradation of polyurethanes

Photodegradation of polyurethanes based on polyester diol-diphenylmethane-p,′p-diisocyanate in the absence and presence of various metal acetylacetonates (Ti, V, Cr, Mn, Co(II), Co(III), Fe, Ni, Cu, Zn, Mg, Al, and Sn) was carried out; and the effect of these metal compounds on the degradation was examined by investigating the changes in the stress–strain properties and infrared- and ultraviolet-visible spectra of the polymer. The results obtained were as follows: Co(III), Co(II), Cu, Ti, and Sn acetylacetonates substantially enhanced the photo-oxidation of the polyurethane; and Al, Ni, and Zn acetylacetonates scarcely affected, or slightly retarded, it. The other metal acetylacetonates accelerated more or less the photo-oxidation of the polymer. Co(III), Co(II), and Cu acetylacetonates extremely degradation. Wavelength of the irradiated light was responsive to photochemical reactions of functional groups in the polymer. C? N and C? O bonds of urethane groups were very accessible to the photo-irradiation both without and with a glass filter, while methylene groups were rather stable to the irradiated light longer than 300 nm, through a glass filter. The effect of the metal compounds on the coloration of the polymer was also consistent with that of mechanical strength and infrared spectral changes.     

Grain refinement of AZ91 alloy by introducing ultrasonic vibration during solidification

Ultrasonic vibration was introduced into the solidification of AZ91 alloy. Various microstructures were produced in this alloy using ultrasonic vibrations at different temperatures of the melt. The coarse dendrite microstructures were obtained with ultrasonic vibrations at temperatures below the liquidus temperature. The fine uniform grains were achieved under ultrasonic vibrations during the nucleation stage, which was mainly attributed to the cavitation and the acoustic flow induced by the ultrasonic vibration.     

Vertically aligned nanowires from boron-doped diamond

Vertically aligned diamond nanowires with controlled geometrical properties like length and distance between wires were fabricated by use of nanodiamond particles as a hard mask and by use of reactive ion etching. The surface structure, electronic properties, and electrochemical functionalization of diamond nanowires were characterized by atomic force microscopy (AFM) and scanning tunneling microscopy (STM) as well as electrochemical techniques. AFM and STM experiments show that diamond nanowire etched for 10 s have wire-typed structures with 3-10 nm in length and with typically 11 nm spacing in between. The electrode active area of diamond nanowires is enhanced by a factor of 2. The functionalization of nanowire tips with nitrophenyl molecules is characterized by STM on clean and on nitrophenyl molecule-modified diamond nanowires. Tip-modified diamond nanowires are promising with respect to biosensor applications where controlled biomolecule bonding is required to improve chemical stability and sensing significantly.     

Chaotic oscillations in Josephson tetrode

We numerically demonstrate the generation of chaos in a four-terminal superconductive device made of five Josephson weak-link junctions, which is referred to as "Josephson tetrode," for the applications of ultrafast random signal generations at frequencies of hundreds of gigahertz. In the Josephson tetrode, two junctions are series-connected and three junctions are parallel-connected. We calculate the dynamics of electrical voltages across the junctions when one of the normal resistances is varied. We confirm the generation of chaos by using a bifurcation diagram, three-dimensional attractors, and the Poincare sections. The bifurcation diagram can be interpreted as the quasi-periodicity-breakdown scenario to chaos. We clarify that the mechanism of the generation of chaos is a nonlinear frequency mixing among three independent voltages across the junctions. The condition of the generation of chaos can be predicted from the values of the coefficients in the equations of our model.