Analysis of the interaction between human plasma fibronectin and gelatin by affinity electrophoresis

The interaction between human plasma fibronectin and gelatin was analyzed by affinity electrophoresis, in which the fibronectin was subjected to electrophoresis in a 4% polyacrylamide gel in the presence and absence of gelatin, as an affinity ligand, and the fibronectin band was stained by an immunoblotting method. The apparent dissociation constants (Kd) of fibronectin for gelatin were calculated from affinity plots based on the original affinity equation at different pHs, urea concentrations, and temperatures. The fibronectin exhibited much lower affinity in the presence of urea. The Kds at 37 degrees C were 1.49 X 10(-7) M, 2.50 X 10(-6) M, and 3.58 X 10(-6) M with 2 M, 3 M, and 4 M urea, respectively. The van't Hoff plots of Kd values against absolute temperature (T) showed that the value of log Kd decreased in proportion to the increase in the value of 1/T within the range of 15-50 degrees C. The standard enthalpy, the standard free energy change at 37 degrees C, and the entropy change at 37 degrees C for association were calculated to be -124.7 kJ/mol, -33.23 kJ/mol, and -295.1 J/mol/deg, respectively. These results suggest that a hydrophilic interaction, such as hydrogen bond or van der Waals interaction, plays an important role in the binding of plasma fibronectin to gelatin.     

Seven-terawatt Ti:sapphire laser system operating at 50 Hz with high beam quality for laser Compton femtosecond X-ray generation

We present a 7-TW Ti:sapphire laser system operating at 50 Hz for laser Compton femtosecond X-ray generation. This laser system delivers 8.4 W of average output power at a repetition rate of 50 Hz with a pulse width of 24 fs. It demonstrates successful management using a dynamically stable resonator in the regenerative amplifier and compensation for thermal lensing by a convex mirror in a ring-type four-pass power amplifier. We also present the results of closed-loop corrections for distorted wavefronts of amplified and compressed laser pulses, using an adaptive optical system consisting of a Shack–Hartmann-type wavefront sensor and a deformable mirror. This closed-loop correction results in dramatic improvements, reducing wavefront distortions below 0.05 λ rms. Received: 31 October 2002 / Revised version: 3 March 2003 / Published online: 5 May 2003 RID="*" ID="*"Corresponding author. Fax: +81-298/61-3349, E-mail: ito@festa.or.jp     

Active synchronization of two mode-locked lasers with optical cross correlation

Two mode-locked Ti:sapphire lasers of different wavelengths were precisely synchronized by a simple feedback system employing sum-frequency generation (cross correlation). When the timing error exceeded the pulse duration, the periodic bunch of the sum-frequency pulse was used for rough timing adjustment. Using cross correlation with a stretched pulse, we struck a balance between wide locking range and sensitive timing detection. When the two lasers were well-synchronized, we obtained a continuous cross-correlation pulse train for 3 min. The holding time of the laser synchronization was extended to over one hour by adding a motorized stage to the PZT-mounted cavity mirror. We estimated the rms timing jitter between the two lasers by a scanning cross-correlation measurement. We confirmed that the rms timing jitter of the two lasers during 1.8 s was 28 fs. Received: 30 January 2002 / Revised version: 14 June 2002 / Published online: 8 August 2002     

Poly(l‐lactic acid)‐modified silica stationary phase for reversed‐phase and hydrophilic interaction liquid chromatography

Poly(l ‐lactic acid) is a linear aliphatic thermoplastic polyester that can be produced from renewable resources. A poly(l ‐lactic acid)‐modified silica stationary phase was newly prepared by amide bond reaction between amino groups on aminopropyl silica and carboxylic acid groups at the end of the poly(l ‐lactic acid) chain. The poly(l ‐lactic acid)‐silica column was characterized in reversed‐phase liquid chromatography and hydrophilic interaction liquid chromatography with the use of different mobile phase compositions. The poly(l ‐lactic acid)‐silica column was found to work in both modes, and the retention of test compounds depending on acetonitrile content exhibited “U‐shaped” curves, which was an indicator of reversed‐phase liquid chromatography/hydrophilic interaction liquid chromatography mixed‐mode retention behavior. In addition, carbonyl groups included into the poly(l ‐lactic acid) backbone work as an electron‐accepting group toward a polycyclic aromatic hydrocarbon and provide π–π interactions.     

Analysis of collagen type III by uninterrupted sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting: changes in collagen type III polymorphism in aging rats.

A new method of type III collagen analysis by uninterrupted sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) combined with immunoblotting was developed. The electrophoresis was carried out with gels containing 4 M urea. A negatively charged reducing agent, thioglycolic acid, was added to the running buffer of the cathodic reservoir between 15 and 20 min after Bromphenol Blue (BPB) migrated to the top of the separating gel, to reduce interchain disulfide binding of the collagen. The polymorphic type III collagens, i.e., an alpha-chain derived from a trimer [alpha 1(III)]3 with interchain disulfide bonds but without covalent cross-links, alpha 1(III), a beta-chain with covalent cross-links, beta(III), or an alpha-chain released from a trimer without reduction of the disulfide bonds, alpha*1(III), were identified by immunostaining and quantified by densitometry. Using this method, changes in collagen type III polymorphism with aging were examined in the aorta, brachial artery, and skin of rats. The total quantity of collagen type III decreased with aging in all tissues. beta(III) was the major component in the aorta and brachial artery, but alpha 1(III) was the major component in the skin. With increasing age from 3 to 60 weeks, the ratio of beta(III) to alpha 1(III), which is correlated with the extent of covalent cross-linking, showed a steep increase in the aorta but only a slight increase in the skin and it remained almost constant in the brachial artery.(ABSTRACT TRUNCATED AT 250 WORDS)     

Fluctuation of femtosecond X-ray pulses generated by a laser-Compton scheme

A short-pulse X-ray-generation experiment was performed by Compton scattering through interaction between a 3-ps electron beam and 100-fs laser photons in a 90° scattering configuration. The observed X-ray intensity was typically 3×10⁴ photons/pulse and roughly matched the theoretically expected intensity. The X-ray energy and pulse duration were estimated theoretically to be 2.3 keV and 280 fs from the observed electron- and laser-beam parameters. The fluctuation of the X-ray output was measured as 25% (rms) during a 30-min operation. The fluctuation was expressed as a function of the fluctuation of the timing between the electron and laser beams. The measured fluctuation of the X-rays was approximately consistent with that caused by the fluctuation of the timing between the beams. Received: 19 November 2001 / Revised version: 23 January 2002 / Published online: 14 March 2002     

Development of a highly stable Yb:YAG thin disk pulsed green laser for high power Ti:sapphire based amplifier at 100 kHz repetition rate

We have developed a pulsed Yb:YAG thin disk green laser for pumping a 100 kHz and 10 W femtosecond laser. We optimized numerically and experimentally the open duration of the acousto-optic (AO) Q-switcher and the internal SHG method, generating stable output energy at a 100 kHz repetition rate. We demonstrated a 515 nm output power of over 80 W with a pulse-to-pulse stability of less than 1% RMS with an LD power to green power conversion efficiency of 24%. With the developed green laser, we obtained 16 W of a Ti:sapphire laser in a cavity dumped mode. PACS 42.55.Xi; 42.60.Gd; 42.60.By; 42.65.-k; 42.65.Re     

Timing jitter in a kilohertz regenerative amplifier of a femtosecond-pulse Ti:Al(2)O(3) laser

We measure the timing error of femtosecond pulses amplified by a Ti:sapphire regenerative amplifier at a 1-kHz repetition rate by use of a modified cross-correlation technique. Linearly frequency-chirped amplified pulses are frequency mixed with transform-limited oscillator pulses. A shift in the sum frequency corresponds to the timing error of each amplified pulse relative to the oscillator pulses. The timing error was measured every 6 ms with approximately 1-fs resolution over a measurable range of 400 fs.     

Tunable mid-infrared,high-energy femtosecond laser source for glyco-protein structure analysis

We have developed a 6–12 μm mid-infrared (MIR) femtosecond laser source for glyco-protein structure analysis. The MIR femtosecond laser pulses are generated by a differential frequency generation (DFG) configuration with a combination of Ti:sapphire based regeneratively amplified femtosecond laser pulses (780 nm, 160 fs, 1 mJ) and a β-BaB₂O₄ (BBO) based optical parametric amplifier (OPA). The MIR pulse energy exceeds 4.5 μJ, where a glyco-protein molecule has resonant absorption lines due to the vibrational–rotational transitions. The pulse width is estimated to be less than 1 ps according to the cross correlation measurement between the two OPA output pulses. Using the MIR femtosecond laser pulses, we demonstrated photo-dissociation of the sialyl Lewis X (sLeX) proton added ion, which is the first time to the best of our knowledge. PACS 42.65.Re; 42.62.-b; 42.60.-b; 42.65.-k; 87.50