A new study of the degradation of hyaluronic acid by hyaluronidase using quartz crystal impedance technique

A new quartz crystal impedance sensing technique for the assay of hyaluronidase (HAse) activity is presented. It is based on the changes in viscosity and density during the enzymatic hydrolysis of hyaluronic acid (HA) by HAse. The variations of equivalent circuit parameters of piezoelectric quartz crystal (PQC) during the enzymatic degradation are discussed. The motional resistance shift curves indicate that the viscosity of the test solutions decreases during the hydrolysis process. The initial hydrolysis rates of HA are obtained from changes in viscosity and density as a function of incubation time. Kinetic parameters (the Michaelis constant K(m) and the maximum hydrolysis rate V(max)) of the degradation reaction are estimated by using a linear Lineweaver-Burk plot in this work. The K(m) was 0.44+/-0.03 mg.ml(-1) and the V(max) was -(5.29+/-0.36)x10(-3) kg.m(-2).s(-1/2).min(-1).     

Combinatorial synthesis of heterocycles: solid-phase synthesis of 6-amino-2,4-dioxo-3,4-dihydro-1,3,5-triazine derivatives

A new solid-phase synthesis of trisubstituted 6-amino-2,4-dioxo-3,4-dihydro-1,3,5-triazines from a resin-bound amine component is described. The amine was readily converted to the corresponding polymer-bound S-methylthiopseudourea, which upon reaction with secondary amines gave the disubstituted guanidines. Cyclization of the polymer-bound guanidines with chlorocarbonylisocyanate afforded the triazinediones. The third point of diversity was introduced by the Mitsunobu reaction. The method is amenable for iterative combinatorial library generation.     

Detection of intact hemoglobin from aqueous solution with laser desorption mass spectrometry

Laser induced liquid beam ionization/desorption mass spectrometry (LILBID-MS) is a new desorption method recently developed in our laboratory. This method allows ions to be desorbed directly from the liquid phase into the high-vacuum region of a mass spectrometer. This method has now been applied to the detection of noncovalent protein-protein complexes. The example given in this paper is the quartenary complex of human hemoglobin. For the first time, the intact hemoglobin could be detected by laser desorption mass spectrometry. Furthermore, evidence for the specificity of the complex is given.     

Additions of enantiopure alpha-sulfinyl carbanions to (S)-N-sulfinimines: asymmetric synthesis of beta-amino sulfoxides and beta-alphamino alcohols

The addition of the lithium anions derived from (R)- and (S)-methyl and -ethyl p-tolyl sulfoxides to (S)-N-benzylidene-p-toluenesulfinamide provides an easy access route to enantiomerically pure beta-(N-sulfinyl)amino sulfoxides. Stereoselectivity can be achieved when the configurations at the sulfur atoms of the two reagents are opposite (matched pair), thus resulting in only one diastereoisomer, even for the case in which two new chiral centers are created. The N-sulfinyl group primarily controls the configuration of the carbon bonded to the nitrogen, whereas the configuration of the alpha-sulfinyl carbanion seems to be responsible for the level of asymmetric induction, as well as for the configuration of the new stereogenic C-SO carbon in the reactions with ethyl p-tolyl sulfoxides. An efficient method for transforming the obtained beta-(N-sulfinyl)amino sulfoxides into optically pure beta-amino alcohols, based on the stereoselective non-oxidative Pummerer reaction, is also reported.     

DIPOLE POLARIZATION RELAXATION AND MOLECULAR STRUCTURE OF MAIN-AND SIDE-CHAIN LIQUID-CRYSTALLINE POLYMERS

Molecular mobility in thermotropic polyesters and side-chain polymers with different struc-ture of mesogens and spacers has been studied by dielectrical method in dilutesolutions. The results made it possible to establish the multiplicity of dielectric relaxationtransitions which reflects the small- and large-scale types of molecular motion. It was shownthat dielectric relaxation processes occurring in accordance with local mechanism (relaxationtimes 10~(-9)--10~(-7)s. and the activation energy 10--50kJ/mol) are due to the mobility of kineticchain elements of different length within a monomer units. It was found that the dielectricrelaxation process connected with a large-scale form of molecular motion (relaxation times10~(-5)--10~(-6)s. and the activation energy 100kJ/mol) did not depend on the molecular massbut was infiuenced by factors changing the conformational state of the macromolecule. It isestablished tha the cooperative reorientation mobility of associated mesogenic fragments isthe source of the large-scale process.     

Characterization of laser-irradiated YNi 2 B 2 C surfaces by Auger electron spectroscopy

The suitability of laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) for precise analysis of YNi(2)B(2)C has been investigated. The intensity ratios B/Y and Ni/Y were found to vary during ablation as a function of the ablation conditions. This could be because of fractionation, owing to incongruent ablation or transport and plasma effects. The bottoms and surroundings of the craters were investigated by scanning Auger electron spectrometry. The bottoms of the craters produced by ablation are covered with a thin oxide comparable with that on the polished crystal surface.The craters are surrounded by an oxide layer the dimensions and thickness of which depend on the laser conditions. The formation of this oxide can be assumed to be a result of partial oxidation of sample material during the ablation process; the oxide is then redeposited around the laser crater.     

From achiral porphyrins to template-imprinted chiral aggregates and further. Self-replication of chiral memory from scratch

The presence of 10-13 M of chiral clusters of aromatic amino acids addresses aggregation of opposite-charged achiral porphyrin towards the formation of smart chiral assemblies. The latter supramolecular complexes are able to self-propagate and transfer their chiral information with a 100% yield. The chiral bias occurs through a correlated sequence of induction, memory, and amplification of chirality that strongly recalls possible prebiotic scenarios.     

Magneto-optics of Ferritin

Measurements of Rayleigh light scattering, nonlinear light scattering in DC magnetic fields, and the Cotton-Mouton effect were carried out for 15 mM NaCl and water solutions of ferritin at room temperature. The spherical geometry of the molecule implies that it is optically isotropic. Such a macromolecule should not manifest magnetic anisotropy; however, in solution it shows induced magnetic birefringence (Cotton-Mouton effect) and changes in the intensity of the scattered light components. The analysis of the obtained results indicates the deformation of linear optical polarizability induced in the ferritin by a magnetic field as the main source of the magneto-optical phenomena observed. Light scattering and the CM effects theoretically depend on the linear magneto-optical polarizability, chi, and the nonlinear magneto-optical polarizability, eta. Using the theory describing the phenomena as well as the experimental data, the values of the anisotropy of linear magneto-optical polarizability components, chi(parallel) - chi(perpendicular) = -(1.3 +/- 0.7) x 10(-22) [cm3] (in SI units chi(parallel) - chi(perpendicular) = -(2.0 +/- 1.2) x 10(-33) [m3]), the linear optical polarizability, alpha = (alpha(parallel) + 2alpha(perpendicular))/3 = (3.9 +/- 1.0) x 10(-20) [cm3] (in SI units alpha = (3.52 +/- 0.09)x10(-4) [Cm2 V(-1)]), and its anisotropy, kappa(alpha) = (alpha(parallel) - alpha(perpendicular))/3alpha = -(0.06+/-0.03), nonlinear magneto-optical polarizability, eta = (eta(parallel) + 2eta(perpendicular))/3 = -(4.7 +/- 0.9) x 10(-30) [cm3 Oe(-2)] (in SI units eta = -(6.7 +/- 1.3) x 10(-18) [Cm4 V(-1) A(-2)]) and its anisotropy, kappa(eta) = (eta[parallel) - eta(perpendicular))/3eta = -(0.15 +/- 0.10), were deduced. Here alpha(parallel), eta(parallel), alpha(perpendicular), eta(perpendicular) are the optical and magneto-optical polarizability components along the parallel and the perpendicular axes of the axially symmetric molecule, respectively.     

Gas chromatographic--mass spectrometric determination of phenylacetic acid in human blood

Phenyl acetic acid, a metabolite of 2-phenyl ethylamine, acts as a neuromodulator in the nigrostriatal dopaminergic pathway stimulating the release of dopamine. The evaluation of phenyl acetic acid concentration in the biological fluid reflects phenyl ethylamine levels thus allowing the assessment of the modulatory role of this endogenous substance. Changes in biological fluids levels of 2-phenylethylamine and/or in its metabolite have been reported in affective disorders, such as depression and schizophrenia. Recently, the occurrence of the "attention deficit hyperactivity syndrome" has been frequently reported in childhood population and involvement of dopaminergic dysfunction in this disease has been suspected. A fast, reliable and reproducible method for the determination of phenyl acetic acid in human blood, is therefore needed in order to have a screening tool for monitoring both healthy childhood population and suspected "attention deficit hyperactivity syndrome" patients. The gas chromatographic-mass spectrometric method here described makes use of a deuterated internal standard in order to overcome problems related to the lack of reproducibility often encountered when a derivatization step is performed.     

The complex of a bivalent derivative of galanthamine with torpedo acetylcholinesterase displays drastic deformation of the active-site gorge: implications for structure-based drug design

Bifunctional derivatives of the alkaloid galanthamine, designed to interact with both the active site of the enzyme acetylcholinesterase (AChE) and its peripheral cation binding site, have been assayed with Torpedo californica AChE (TcAChE), and the three-dimensional structures of their complexes with the enzyme have been solved by X-ray crystallography. Differences were noted between the IC(50) values obtained for TcAChE and those for Electrophorus electricus AChE. These differences are ascribed to sequence differences in one or two residues lining the active-site gorge of the enzyme. The binding of one of the inhibitors disrupts the native conformation of one wall of the gorge, formed by the loop Trp279-Phe290. It is proposed that flexibility of this loop may permit the binding of inhibitors such as galanthamine, which are too bulky to penetrate the narrow neck of the gorge formed by Tyr121 and Phe330 as seen in the crystal structure.