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Biomedical subjects

M Scandola

Publications and source records attributed to M Scandola.

12 recordsLinked to original sources

A new type of thermoalkalophilic hydrolase of Paucimonas lemoignei with high specificity for amorphous polyesters of short chain-length hydroxyalkanoic acids.

A novel type of hydrolase was purified from culture fluid of Paucimonas (formerly Pseudomonas) lemoignei. Biochemical characterization revealed an unusual substrate specificity of the purified enzyme for amorphous poly((R)-3-hydroxyalkanoates) (PHA) such as native granules of natural poly((R)-3-hydroxybutyrate) (PHB) or poly((R)-3-hydroxyvalerate) (PHV), artificial cholate-coated granules of natural PHB or PHV, atactic poly((R,S)-3-hydroxybutyrate), and oligomers of (R)-3-hydroxybutyrate (3HB) with six or more 3HB units. The enzyme has the unique property to recognize the physical state of the polymeric substrate by discrimination between amorphous PHA (good substrate) and denatured, partially crystalline PHA (no substrate). The pentamers of 3HB or 3HV were identified as the main products of enzymatic hydrolysis of native PHB or PHV, respectively. No activity was found with any denatured PHA, oligomers of (R)-3HB with five or less 3HB units, poly(6-hydroxyhexanoate), substrates of lipases such as tributyrin or triolein, substrates for amidases/nitrilases, DNA, RNA, casein, N-alpha-benzoyl-l-arginine-4-nitranilide, or starch. The purified enzyme (M(r) 36,209) was remarkably stable and active at high temperature (60 degrees C), high pH (up to 12.0), low ionic strength (distilled water), and in solvents (e.g. n-propyl alcohol). The depolymerase contained no essential SH groups or essential disulfide bridges and was insensitive to high concentrations of ionic (SDS) and nonionic (Triton and Tween) detergents. Characterization of the cloned structural gene (phaZ7) and the DNA-deduced amino acid sequence revealed no homologies to any PHB depolymerase or any other sequence of data banks except for a short sequence related to the active site serine of serine hydrolases. A classification of the enzyme into a new family (family 9) of carboxyesterases (Arpigny, J. L., and Jaeger, K.-E. (1999) Biochem. J. 343, 177-183) is suggested.

Amino Acid Sequence↗

Natural cellulose fibers: heterogeneous acetylation kinetics and biodegradation behavior.

Steam-exploded fibers from flax (Linum usitatissimum) are heterogeneously acetylated using acetic anhydride and sulfuric acid as catalyst, with the aim to modify the surface properties without changing fiber structure and morphology. The acetylation reaction follows first-order kinetics up to a reaction time that depends on catalyst concentration (15 h when using 0.4 vol % of H(2)SO(4) or 50 h with 0.1 vol %). The fibers undergo no structural and/or morphological changes under either reaction condition. On the contrary, surface damage and structural modifications appear after longer reaction times, when the reaction kinetics change. The extent of biodegradation of acetylated fibers, evaluated from the weight percent remaining after 13 days of exposure to previously isolated cellulolytic bacteria Cellvibrio sp., decreases with increasing acetylation degree. After biodegradation the fibers show a higher acetyl content than before the experiment, indicating that the bacteria preferentially biodegrade unsubstituted cellulose, though also acetylated chains are cleaved. Biodegradable acetylated cellulose fibers with modified surface chemistry and unchanged structure are obtained for applications as polymer composite reinforcements.

Acetylation↗

Bacterial poly(3-hydroxybutyrate): an optical microscopy and microfocus X-ray diffraction study.

Two-dimensional spatially resolved microfocus X-ray diffraction has been used to investigate spherulites of pure bacterial poly(3-hydroxybutyrate) (PHB) and of a blend of natural and synthetic atactic PHB (a-PHB) crystallized at a relatively high temperature (Tc = 140 degrees C). Both samples investigated contained practically two-dimensional spherulites, characterized by wide extinction bands (band spacing > 80 microns). The X-ray diffraction patterns confirmed that the unit cell alpha-axis is oriented along the spherulite radius in PHB and that the same is true for the a-PHB containing blend. Comparison of the matrix of diffraction patterns with the polarized optical micrograph of the scanned sample area indicated a very clear correlation between pattern changes and banding, yielding a straightforward picture of the structural variations within the spherulite.

Bacteria↗

Viscoelastic and thermal properties of collagen/poly(vinyl alcohol) blends.

Blends of poly(vinyl alcohol) (PVA) with collagen and gelatin, prepared from aqueous solution by solvent casting, were investigated by differential scanning calorimetry (DSC) and dynamic mechanical thermal analysis (DMTA). After conditioning at 51% relative humidity, collagen and PVA show nearly coincident glass transition temperatures (Tg approximately 35 degrees C), while gelatin has a higher Tg (approximately 70 degrees C). Gelatin/PVA blends show two invariant Tgs, whose temperature and associated specific heat increment clearly indicate the coexistence of two amorphous phases composed of the pure components. Owing to similarity of the Tgs of collagen and PVA after humidity conditioning, DSC offers no indication on miscibility of collagen/PVA blends. In DMTA experiments, where absorbed water freely evaporates from the samples during the thermal scan, PVA shows a glass transition relaxation at about 50 degrees C, while both gelatin and collagen display an intense glass transition in the vicinity of 230 degrees C. The DMTA spectra of collagen/PVA and gelatin/PVA blends show two invariant glass transition relaxations at about 50 and 230 degrees C. Absence of any Tg shift with composition demonstrates that the blend components are immiscible. However, blends of PVA with collagen and gelatin form optically clear films with good mechanical properties over the whole range of compositions. It is found that at T > Tg (PVA) the elastic modulus (E') of the blends strongly increases with increasing content of the biopolymer. In the case of collagen/PVA blends, experimental E' values agree with the predictions of a simple two-phase composite model with phases connected in parallel. It is concluded that, though thermodynamically immiscible with both native and denatured collagen, PVA forms mechanically compatible blends with collagen and gelatin.

Animals↗

Qualitative analysis of potential metabolites and degradation products of a new antiinfective drug in rat urine, using HPLC with radiochemical detection and HPLC-mass spectrometry.

High-performance liquid chromatography (HPLC) was used in combination with radioactivity detection and mass spectrometry (MS) to elucidate the metabolic fate of GV104326, a novel tricyclic beta-lactam antibacterial agent. Metabolic profiles were obtained by analysis of rat urine samples collected after intravenous administration of the 14C-labelled drug at the dose of 50 mg kg-1 (12.95 MBq kg-1). Methods for solid-phase extraction from urine samples and for reversed-phase chromatographic separation of drug related material were developed. HPLC-MS was used to confirm that the parent compound corresponded to the principal peak in the chromatograms, and two minor peaks were identified as potential metabolites of GV104326. They were shown to be an open beta-lactam ring derivative (GV173923) and a dimeric compound (GV196359).

Animals↗

Molecular motions of polysaccharides in the solid state: dextran, pullulan and amylose.

Dextran, pullulan and amylose have been investigated by differential scanning calorimetry, thermogravimetric analysis, dynamic mechanical and dielectric spectroscopy over a wide range of temperatures and frequencies. No melting or glass transition is seen below the range of thermal degradation (about 300 degrees C) for either amylose or pullulan; only dextran shows a Tg at 223 degrees C (delta cp = 0.40 J/g deg). The viscoelastic spectrum of the 'dry' polysaccharides is characterized by a low temperature relaxation that occurs at -94, -73 and -59 degrees C, at 1 kHz, (activation energy 32, 39 and 52 kJ/mol) in dextran, pullulan and amylose respectively and is assigned to small entity local motions of the polysaccharide backbone. Absorbed water strongly modifies the relaxation spectrum, inducing a new relaxation below room temperature and dissipation regions associated with water loss above room temperature. The former appears at temperatures higher than the relaxation characteristic of the dry polymer and moves to lower temperature with increasing water content. In normal 'room humidity' conditions (about 10% absorbed water) the water-induced relaxation, attributed to the motion of complex polymer-water relaxing units, is the only observable feature in the dynamic mechanical and dielectric spectrum below room temperature.

Amylose↗

Viscoelastic relaxations and thermal properties of bacterial poly(3-hydroxybutyrate-co-3-hydroxyvalerate) and poly(3-hydroxybutyrate-co-4-hydroxybutyrate).

3-Hydroxybutyrate-3-hydroxyvalerate (3HB-3HV) as well as 3-hydroxybutyrate-4-hydroxybutyrate (3HB-4HB) copolyesters have been investigated by differential scanning calorimetry, thermogravimetric analysis and dynamic mechanical spectroscopy, over a wide range of compositions (0-95 mol% 3HV; 0-82 mol% 4HB). Both series of isolated copolyesters are partially crystalline at all compositions. Quenched samples show a glass transition that decreases linearly with increasing co-monomer molar fraction, more markedly when the co-monomer is 4HB. Above Tg, all copolyesters, rich in 3HB units, show a cold crystallization phenomenon followed by melting, while at the other end crystallization on heating is observed only in 3HB-3HV copolymers. The viscoelastic spectrum, strongly affected by thermal history, shows two relaxation regions: the glass transition, whose location depends on copolymer type and composition, and a secondary dispersion region at low temperatures (-130/-80 degrees C). The latter results from a water-related relaxation analogous to that of P(3HB) and, in 3HB-4HB copolymers, from another overlapping absorption peak centered at -130 degrees C, attributed to local motion of the methylene groups in the linear 4HB units.

Calorimetry, Differential Scanning↗

Mass spectrometric approaches in structural characterization of cephalosporins.

The mass spectrometric behaviour of five structurally different cephalosporins has been studied in detail by means of different ionization/desorption methods (electron impact, fast atom bombardment, desorption chemical ionization, laser-induced vaporization) and metastable ion studies (linked scans and mass-analysed ion kinetic energy spectrometry). The best results were obtained by fast atom bombardment mass spectrometry, leading to both molecular ions and fragment ions diagnostic for structural identification.

Cephalosporins↗

Calorimetric studies of elastin solvation.

The interaction of ox ligamentum nuchae elastin (native, purified and soluble) with four different solvents was investigated by Differential Scanning Calorimetry. The freezable solvent content was determined, at different total solvent contents, from the melting endotherms, and the amount of unfreezable solvent was obtained by extrapolation. For two bifunctional solvents, water and ethylene glycol, the molar ratio "bound solvent/elastin residue" was the same for all the elastin samples investigated, and very close to 1.5. This result is identical to that obtained by other workers for the "water/CO-NH group" of synthetic amorphous polyamides, and it can suggest that the hydration of elastin is interpretable on the basis of a similar model. The model is based on a direct interaction of water with the peptide group, and suggests that 0.5 moles of water are strongly bound, and 1.0 moles are loosely bound to the CO-NH group. The ratio "bound water/elastin residue" is larger for the soluble beta elastin. Different molar ratios are also obtained for monofunctional solvents such as methanol and trifluoroethanol.

Animals↗