Adsorption of detergent-solubilized and phospholipase C-solubilized alkaline phosphatase at air/liquid interfaces

To investigate the influence of a hydrophobic anchor on protein adsorption, equilibrium and dynamic aspects of the adsorption of two different solubilized forms of rat osseous plate alkaline phosphatase on Langmuir monolayers of dimyristoylphosphatidic acid (DMPA) were studied. Surface pressure and surface potential measurements at air/liquid interfaces were carried out using the detergent-solubilized form (DSAP) of alkaline phosphatase, which holds a glycosylphosphatidylinositol (GPI) hydrophobic anchor, and the glycosylphosphatidylinositol-specific phospholipase C-solubilized form (PLSAP), lacking the GPI anchor. Similar surface transitions observed for both DMPA and DMPA/PLSAP mixed monolayers indicate that the presence of PLSAP does not promote significant changes in surface packing of the DMPA monolayer. However, PLSAP interacts with the polar portion of the phospholipid even at high lateral compression. The presence of the GPI anchor increases the adsorption of DSAP at a plain air/liquid interface and also enables the penetration of the protein into the DMPA monolayers. The penetration is dependent on both time and surface pressure. Up to 20 mN/m, the surface pressure increases smoothly indicating a diffusion followed by an adsorption process. Above 20 mN/m, after a fast increase, the surface pressure slowly decays to equilibrium values quite close to the initial surface pressures. The results indicate that the molecular packing of the lipid layer drives the enzyme adsorption to the interface either through the GPI anchor or by the polypeptide moiety.     

Baryon pairs production ine + e − annihilation at\sqrt s = 2.4 GeV

Search for baryon pairs production ine ⁺ e ^? annihilation at \(\sqrt s = 2386 MeV\) is reported. The data relate to a luminosity of 161 nb^?1 collected by the DM2 experiment at DCI, the Orsay colliding ring. First measurements of directe ⁺ e ^? annihilation into \(\Lambda \bar \Lambda \) and ofe ⁺ e ^?→ \(p\bar p\) at this energy are presented. First observation of a goode ⁺ e ^?→ \(n\bar n\) candidate is reported and upper limits are given fore ⁺ e ^?→ \(n\bar n, \Lambda \bar \sum ^0 + c.c.\) and \(\Sigma ^0 \bar \Sigma ^0 \) .     

Two-colour operation of a Free-Electron Laser and applications in the mid-infrared

The two-colour operation of a Free-Electron Laser (FEL) has been demonstrated with the “CLIO” infrared facility. A two section undulator allows the production of picosecond laser pulses at two different wavelengths and simultaneously, in a wavelength range from to , and with a wavelength separation of two colours of up to . The time overlap, between both colours, has been measured on a picosecond time scale and on a microsecond time scale. An initial pump-probe application experiment has been performed with the two colours: stimulated emission has been measured in a 3-level Quantum-Well system. This is the best demonstration of the stability and reliability of the two-colour laser operation. Received: 4 November 1997 / Revised in final form: 31 March 1998 / Accepted: 10 April 1998     

Long polaron lifetime in InAs/GaAs self-assembled quantum dots

We have investigated the polaron dynamics in n-doped InAs/GaAs self-assembled quantum dots by pump-probe midinfrared spectroscopy. A long T1 polaron decay time is measured at both low temperature and room temperature, with values around 70 and 37 ps, respectively. The decay time decreases for energies closer to the optical phonon energy. The relaxation is explained by the strong coupling for the electron-phonon interaction and by the finite lifetime of the optical phonons. We show that, even for a large detuning of 19 meV from the LO photon energy in GaAs, the carrier relaxation remains phonon assisted.     

Long-acting androgens: Analytical and preparative HPLC of testosterone esters

Procedures are described for the normal phase and reverse phase HPLC analysis of testosterone esters having diverse side chains: examples are given of the gram-scale purification of these compounds using two commercial preparative liquid chromatography and an economical and efficient laboratory-assembled preparative liquid chromatograph.     

Vibrational Spectroscopy as a Promising Tool to Study Enzyme-Carrier Interactions: A Review

Abstract

The high selectivity and specificity together with the catalytic activity of enzymes lead them to be a key tool in biotechnology industries. The enzyme immobilization on a carrier material facilitates the reuse of the enzyme and improves its stability. This article is a comprehensive review that reports papers in which different enzymes have been immobilized on distinct carrier materials and enzyme-carrier interactions were evaluated by infrared and/or Raman spectroscopy. Vibrational spectroscopy was used to inspect the spectral bands of different classes of enzymes before and after their immobilization on carriers. In fact, the characteristic spectral bands that prove the interaction enzyme-carrier are related with the amide functional groups of enzymes. In addition, those interactions are characterized by the shift, broadening, or increment on intensity of specific bands in reference to the spectrum of their carrier material. In this review, 20 of these studies used infrared spectroscopy and only three used Raman spectroscopy. The use of both vibrational spectroscopy techniques is increasingly due to their proved capacity to characterize the enzyme-carrier interactions without damaging them.     

The role of probe–probe interactions on the hybridization of double-stranded DNA targets onto DNA-modified magnetic microparticles

In this work, we have studied the effect of different probe lengths and surface densities on the hybridization of a 181-bp polymerase chain reaction product to probes tethered onto magnetic microparticles. Hybridization was shown to be favored by longer probes but only at probe surface densities where probe-to-probe interactions are absent. From these results, a simple rule was inferred for determining maximum surface densities above which hybridization signals decreased. According to this rule, if the average surface area occupied by an immobilized probe (Σ) is larger than the projected surface area of each tethered probe molecule (S _ss ), hybridization efficiency increases with surface density, whereas the reverse occurs when Σ −  S _ss  < 0.     

Hydrophobic interaction chromatography of homo-oligonucleotides on derivatized Sepharose CL-6B. Application of the solvophobic theory

This work describes the hydrophobic interaction chromatography of homo-deoxyoligonucleotides polyA, polyT and polyU with sizes up to 30 bases on a Sepharose gel derivatized with the 1,4-butanediol diglycidyl ether. The oligonucleotides interacted differently with the column according to the molecular mass, the hydrophobic character of the individual bases, the secondary structure of the molecule and the concentration of ammonium sulphate in the eluent. The retention factor, k', was determined from the chromatographic profiles at different concentrations of ammonium sulphate. A linear relationship between log k' and the concentration of ammonium sulphate in the eluent was found for all oligonucleotides at the higher concentrations (> 1.0 M) of ammonium sulphate. The slope of these plots, termed the hydrophobic interaction parameter, was found to be an increasing function of the number of nucleotides. The same plots reveal that polyA molecules with high molecular mass have lower retention factors when compared with polyT, an observation that was not expected since the hydrophobicity of adenine is higher than that of thymine. This behaviour was due to the existence of secondary structures in polyA, which decrease the exposed hydrophobic area of the molecule.     

Sub-100 nm IR spectromicroscopy of living cells

We have performed IR spectromicroscopy of cells immersed in liquid water, with a lateral resolution better than 100 nm. Here, we use the motion of an atomic force microscope tip, probing the local transient deformation induced by an IR pulsed laser tuned at a sample absorbing wavelength. By Fourier analysis of the vibration of the cantilever tip, we can discriminate frequencies that are characteristic of the object, thus eliminating the influence of the water absorption. This opens the door of chemical imaging of living species in vivo, with spatial resolution of the order of the size of cell components.