Comparison of Sepharose CL-2B and CL-4B Matrices in Purification of the Hepatitis B Surface Antigen by Affinity Chromatography

Sepharose CL-4B and Sepharose CL-2B with immobilized CB.Hep-1 monoclonal antibody were compared for studying the matrix efficiency in the immunopurification of the recombinant hepatitis B surface antigen (rHBsAg). The elution capacities of both matrices were similar over eight chromatographic cycles, significant differences were only observed in the first purification cycles. A high percentage of the adsorbed rHBsAg was not eluted from both matrices. The rHBsAg purity was not affected by matrix characteristics (pore size and percentage of agarose) and differences between Sepharose CL-4B and Sepharose CL-2B do not provoke differences in the antibody released from the matrices under defined experimental conditions.     

Probing the Bioactive Conformation of an Archetypal Natural Product HDAC Inhibitor with Conformationally Homogeneous Triazole‐Modified Cyclic Tetrapeptides

Fooling enzymes with mock amides : Analogues of apicidin, a cyclic‐tetrapeptide inhibitor of histone deacetylase (HDAC), were designed with a 1,4‐ or 1,5‐disubstituted 1,2,3‐triazole in place of a backbone amide bond to fix the bond in question in either a trans‐like or a cis‐like configuration. Thus, the binding affinity of distinct peptide conformations (see picture) could be probed. One analogue proved in some cases to be superior to apicidin as an HDAC inhibitor.

     

Comparison of stir bar sorptive extraction and membrane-assisted solvent extraction as enrichment techniques for the determination of pesticide and benzo[a]pyrene residues in Brazilian sugarcane juice

The biodegradation of the organophosphorus insecticide methyl parathion (MP) in aqueous environment by bacteria isolated from river sediment has been studied. Two species of bacteria which show strong MP degradation ability are identified as Shewanella and Vibrio parahaemolyticus. The biodegradation of MP proceeded rapidly with the formation of a series of intermediate products, which were analyzed using a combination of GC/MS and HPLC/ESI-TOFMS techniques. The major products tentatively identified include a series of reduced products of MP. Results demonstrate that the coupling of TOFMS to HPLC enhances further the capability of LC-MS in the identification of polar organic species in complex environmental samples. Degradation pathways leading to the formation of these products are proposed which involves first the reduction of nitro to amino group in MP, followed by combination with some intrinsic matters of bacteria. The mechanism and products from biodegradation are quite different from those of photocatalytic process for which the main intermediates included methyl paraoxon and 4-nitrophenol.     

75th birthday of Professor Leonid Isaakovich Belen’kii

Anniversaries and Dates

75th birthday of Professor Leonid Isaakovich Belen’kii     

Bicyclo[3.2.1]octanone catalysts for asymmetric alkene epoxidation: the effect of disubstitution

A series of 2-fluoro-8-oxabicyclo[3.2.1]octan-3-ones are prepared and tested as catalysts for alkene epoxidation with Oxone^®. These catalysts provide trans-stilbene oxide with up to 83% ee, but the highest ee value is obtained with the monofluorinated ketone 2: both 2,2- and 2,4-disubstituted catalysts afford epoxide of lower ee.     

Diffraction aperture non-ideal behaviour of air coupled transducers array elements designed for NDT

Air coupled piezoelectric ultrasonic array transducers are a novel tool that could lead to interesting advances in the area of non-contact laminar material testing using Lamb wave's propagation techniques. A key issue on the development of such transducers is their efficient coupling to air media (impedance mismatch between the piezoelectric material and air is 90 dB or more). Adaptation layers are used in order to attain good matching and avoid possible serious signal degradation. However, the introduction of these matching layers modify the transducer surface behaviour and, consequently, radiation characteristics are altered, making the usual idealization criteria (of uniform surface movement) adopted for field simulation purposes inaccurate. In our system, we have a concave linear-array transducer of 64 elements (electrically coupled by pairs) working at 0.8 MHz made of PZ27 rectangular piezoceramics (15 mm x 0.3 mm) with two matching layers made of polyurethane and porous cellulose bonded on them. Experimental measurements of the acoustic aperture of single excited array elements have shown an increment on the geometrical dimensions of its active surface. A sub-millimeter vibrometer laser scan has revealed an extension of the aperture beyond the supposed physical single array element dimensions. Non-uniform symmetric apodized velocity surface vibration amplitude profile with a concave delay contour indicates the presumed existence of travelling wave phenomena over the surface of the outer array matching layer. Also, asymptotic propagation velocities around 2500 m/s and attenuation coefficient between 15 and 20 dB/mm has been determined for the travelling waves showing clear tendencies. Further comparisons between the experimental measurements of single array element field radiation diagram and simulated equivalent aperture counterpart reveal good agreement versus the ideal (uniform displaced) rectangular aperture. For this purpose an Impulse Response Method (IRM) has been used.     

Self-assembly of a catechol-based macrocycle at the liquid-solid interface: experiments and molecular dynamics simulations

This combined experimental (STM, XPS) and molecular dynamics simulation study highlights the complex and subtle interplay of solvent effects and surface interactions on the 2-D self-assembly pattern of a Schiff-base macrocycle containing catechol moieties at the liquid-solid interface. STM imaging reveals a hexagonal ordering of the macrocycles at the n-tetradecane/Au(111) interface, compatible with a desorption of the lateral chains of the macrocycle. Interestingly, all the triangular-shaped macrocycles are oriented in the same direction, avoiding a close-packed structure. XPS experiments indicate the presence of a strong macrocycle-surface interaction. Also, MD simulations reveal substantial solvent effects. In particular, we find that co-adsorption of solvent molecules with the macrocycles induces desorption of lateral chains, and the solvent molecules act as spacers stabilizing the open self-assembly pattern.     

Determination of lipoic acid, Trolox methyl ether and tocopherols in human plasma by liquid-chromatography and ion-trap tandem mass spectrometry

A method for the simultaneous determination of lipoic acid and/or Trolox methyl ether, along with α‐, γ‐ and δ‐tocopherol was developed using liquid chromatography–tandem mass spectrometry with negative electrospray ionization (HPLC‐ESI‐MS/MS) in an ion‐trap mass spectrometer. Detection and quantification were accomplished by a multiple reaction monitoring method, using specific transitions from precursor ion to product ion for each analyte. Chromatographic separation was achieved in a 12 min run using a C₁₈‐bonded phase and methanol–aqueous ammonium acetate elution gradient. Linear correlations of the chromatographic peak area (r.u. × s^?1) to the injected amount (ng) gave the slope values (r.u. × s^?1 × ng^?1) 2.34 × 10⁴ for α‐tocopherol, 5.05 × 10⁴ for γ‐tocopherol, 1.27 × 10⁵ for δ‐tocopherol, 8.86 × 10⁵ for lipoic acid and 1.23 × 10⁵ for Trolox methyl ether. The lower limit of quantification ranged between 0.02 and 1.22 ng for Trolox methyl ether and lipoic acid. MS³ experiments of γ‐ and δ‐tocopherol suggest ion‐radical reactions and dependence of the tocopherol fragmentation pattern on the phenolic ring methylation degree. The method is shown to be applicable to measurement of these metabolites in human serum after extraction. Copyright © 2012 John Wiley & Sons, Ltd.     

Ultrasonic velocity and amplitude characterization of magnetorheological fluids under magnetic fields

Variations in velocity of sound and amplitude of the signal of a commercial magnetorheological fluid under different magnetic fields are studied experimentally. Different factors such as orientation, uniformity, geometry and intensity of the magnetic field are investigated. An increase in the change of MR fluid acoustical properties is obtained when the magnetic field intensity is risen. In addition, these properties show an opposite behavior when a magnetic field is applied parallel or perpendicular to the ultrasound propagation. Experiments using an electromagnet and permanent magnets as the source of magnetic field are also compared. Properties such as anisotropy in sound velocity and amplitude make these materials interesting regarding applications.     

Ultrafast photophysics of the isolated indole molecule

The relaxation dynamics of the isolated indole molecule has been tracked by femtosecond time-resolved ionization. The excitation region explored (283-243 nm) covers three excited states: the two ππ* L(b) and L(a) states, and the dark πσ* state with dissociative character. In the low energy region (λ > 273 nm) the transients collected reflect the absorption of the long living L(b) state. The L(a) state is met 1000-1500 cm(-1) above the L(b) origin, giving rise to an ultrafast lifetime of 40 fs caused by the internal conversion to the lower L(b) minimum through a conical intersection. An additional ~400 fs component, found at excitation wavelengths shorter than 263 nm, is ascribed to dynamics along the πσ* state, which is likely populated through coupling to the photoexcited L(a) state. The study provides a general view of the indole photophysics, which is driven by the interplay between these three excited surfaces and the ground state.