VDUP1 potentiates Ras-mediated angiogenesis via ROS production in endothelial cells.

Vitamin D3 up-regulated protein 1 (VDUP1) is a tumor suppressor of which expression is reduced in a variety of cancer cells, and enforced expression inhibits the tumor cell proliferation. It inhibits the activity of thioredoxin, thus contributing cellular ROS generation. Since ROS is a critical factor for angiogenesis, we investigated the role of VDUP1 in angiogenesis and endothelial proliferation. The expression of VDUP1 was upregulated by overexpression of an oncogene, Ras. Enforced expression of VDUP1 increases ROS production and proliferation of Ras-overexpressing endothelial cells. Overexpression of VDUP1 increases the resistance to the anchorage-dependent cell death and tube formation of the Ras-overexpressing endothelial cell. In addition, the removal of ROS by ROS scavenger attenuates the effect of VDUP1 on tube formation. These results suggest that VDUP1 is involved in Ras-mediated angiogenesis via ROS generation in endothelial cells.

Importance of phosphatidylethanolamine for the interaction of apocytochrome c with model membranes containing phosphatidylserine.

The effect of phosphatidylethanolamine (PE) on the binding of apocytochrome c to model membranes was examined. When 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) of the standard vesicles composed of 80% of this lipid and 20% of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoserine (POPS) was gradually replaced with upward of 50% of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine (POPE), the binding increased appreciably. Ca(2+), causing the phase separation of PS, also brought about increased binding of apocytochrome c in the PC/PS system, underlining the importance of PS properties in membranes for the protein binding. The resonance energy transfer between Trp-59 in apocytochrome c and pyrene-PS incorporated into bilayers showed that the replacement of PC with PE increased the extent of apocytochrome c penetration into membranes by a PE concentration-dependent manner. However, in the absence of PS, PE had no apparent effect on these functions of apocytochrome c, suggesting that PE-induced change(s) of acidic membrane properties is important to the association of apocytochrome c with vesicles. From the observations that the excimer to monomer fluorescence ratio of pyrene-PS increased and the fluorescence of NBD-PS was quenched with increasing concentration of PE, it was deduced that PE caused PS-enriched domains in PC/PE/PS membranes. The colocalization of pyrene-PS with BODIPY-PS by PE further supported the possibility. We suggest that PE-induced formation of PS-enriched domains acts as binding sites for apocytochrome c in membranes.

High-sensitivity detection of CH radicals in flames by use of a diode-laser-based near-ultraviolet light source.

CH radical distributions in ethylene-air and methane-air diffusion flames are mapped by wavelength-modulation absorption spectroscopy (WMS). Tunable, wavelength-modulated 426-nm light is generated by frequency doubling of a modulated 852-nm distributed Bragg reflector diode laser. Absorbances of 5 x 10(-5) are measured with second-harmonic (2f) WMS with a signal-to-noise ratio of 3:1 in a 3-Hz measurement bandwidth. The feasibility of simultaneous line-of-sight absorption and spatially resolved laser-induced-fluorescence detection with a single excitation beam is also demonstrated. This near-UV source is suitable for microgravity drop-tower experiments and other applications in which compact, rugged, energy-efficient instrumentation is required.

Frequency Modulation Multiplexing for Simultaneous Detection of Multiple Gases by use of Wavelength Modulation Spectroscopy with Diode Lasers.

Modulation frequency multiplexing provides a straightforward method, analogous to television or radio broadcasting, for performing simultaneous detection of multiple gases by use of wavelength modulation spectroscopy with diode lasers. When fiber-optic coupled lasers are used, our approach guarantees that all beams transit the same optical path and impinge on the same detector. Each laser is modulated at a different frequency and the detector output is processed by a set of lock-in amplifiers, one for each laser, to measure the absorbance encountered by each laser.

Time-resolved wavelength modulation spectroscopy measurements of HO 2 kinetics.

High-frequency wavelength modulation spectroscopy (WMS) has been applied to the detection of the hydroperoxyl radical (HO2 ) in a laser photolysis and long-path absorption pump-probe kinetics reactor with a near-infrared distributed feedback diode laser. The HO2 is formed by the 355-nm photolysis of Cl2 in the presence of CH3 OH and O2 and monitored by a phase-sensitive detection of the second-harmonic (2f ) signal in the 2?1 band with a 1.5- ?m diode laser directly modulated at 5 MHz. The measured 2f WMS signal is calibrated by direct absorption and converted to an absolute number density with the known absorption line strength of the HO2 line at 6625.80cm-1 . The utility of time-resolved WMS as a second-order kinetics probe is demonstrated through the measurement of the HO2 self-reaction rate constant at 295 K.

Measurement of nitric oxide with an antimonide diode laser.

An antimonide diode laser operating near 2.65 mum was used to measure absorption lines of NO gas in the first overtone band. A blended line pair of NO that is sufficiently free of interference from H(2) O to permit the selective detection of NO under reduced pressure conditions was identified. With wavelength-modulation spectroscopy, a rms noise level equivalent to an absorbance of 3.2 x 10(-5) was achieved at a measurement integration time (for a single spectral data point) of 0.1 s. The corresponding detection sensitivity (signal-to-noise ratio of 2) for NO in air at reduced pressure was ~15 ppm m (ppm is parts in 10(6)). Antimonide diode lasers show substantial promise for gas-sensing applications because they can gain access to relatively strong absorption lines of several gases of environmental interest at operating wavelengths at which cryogenic cooling is not required.

Structure of a methionine-rich segment of Escherichia coli Ffh protein.

The methionine-rich segments of the Ffh protein of Escherichia coli and its eukaryotic counterpart SRP54 are thought to bind signal sequences of secretory proteins. The structure of a chemically synthesized 25-residue-long peptide corresponding to one of the proposed methionine-rich amphiphilic helices of Ffh was determined in water and in aqueous trifluroethanol (TFE) solution using CD and NMR. An appreciable alpha-helix conformation exists even in water and this peptide assumes a stable alpha-helix along most of its length in aqueous TFE solution. It is clear that this segment of Ffh protein has a very strong propensity to form alpha-helical structure.

Diode-laser-based sum-frequency generation of tunable wavelength-modulated UV light for OH radical detection.

Greater than 1.5 microW of tunable wavelength-modulated 308-nm radiation was generated by sum-frequency mixing of 90 mW of 835-nm diode-laser output and 1.5 W of 488-nm Ar(+)-laser output in a beta-barium metaborate crystal. Hydroxyl radicals formed in a discharge-flow reactor were detected by use of the generated UV beam by direct absorption, wavelength-modulation absorption spectroscopy, and laser-induced fluorescence. Wavelength modulation with second-harmonic detection yielded an estimated minimum detectable absorbance of 8.5 x 10(-6) at a 1-Hz bandwidth. This absorbance level corresponds to an OH detection sensitivity of 11.7 parts per trillion for a 1-m path length at 1 atm and 298 K.

Wavelength-modulation detection of acetylene with a near-infrared external-cavity diode laser.

An external-cavity diode laser operating at 1500 nm was used to record the combination band of acetylene (C(2)H(2)). By combination of wavelength-modulation spectroscopy with a noise-canceler detection circuit, a minimum detectable absorbance of 4.8 × 10(-4) with a 300-ms time constant was achieved, although this result was limited by étalon fringes. When combined with this detection technique, continuous, widely tunable output from an external-cavity laser is ideally suited for high-resolution absorption spectroscopy with excellent sensitivity.