Peptide labeling using 188Re, 188Re-MAG3 and 153Sm-H1ETA: A comparison on their in vitro lipophilicity

Lanreotide peptide was labeled with ¹⁵³Sm-H₁ETA and ¹⁸⁸Re-MAG₃ in order to evaluate whether or not their conjugation to the peptide produce significant differences of the in vitro lipophilicity with respect to the ¹⁸⁸Re-lanreotide prepared by the direct labeling method (highly lipophilic). The differences of lipophilicity between the complexes, were evaluated using a reverse phase HPLC system. The measured lipophilicity of ¹⁵³Sm-H₁ETA-lanreotide, ¹⁸⁸Re-MAG₃-lanreotide and ¹⁸⁸Re-lanreotide was taken to be the capacity factor [k" = (t _R-t ₀)/t ₀ where t _R is the retention time and t ₀ is the dead time] for each of the complexes under identical chromatography conditions. Results showed that the in vitro lipophilicity decreased in the order ¹⁸⁸Re-lanreotide (direct labeling), ¹⁸⁸Re-MAG₃-lanreotide and ¹⁵³Sm-H₁ETA-lanreotide. Since the last one has a capacity factor (k") similar to that of ¹⁸⁸Re-MAG₃, some renal elimination for ¹⁵³Sm-H₁ETA-lanreotide could be expected, which probably would reduce the unnecessary radiation dose to normal tissues.     

Bis(amino acid) derivatives of 1,4-diamino-2-butyne that adopt a C2-symmetric turn conformation

1,4-Diamino-2-butyne was prepared from 1,4-dichloro-2-butyne via 1,4-diazido-2-butyne. Bis(amino acid) derivatives of 1,4-diamino-2-butyne having the general structure (Boc-Xxx-NHCH2C[triple bond])2 (Xxx = Ala, Phe and Met) were prepared and examined by 1H NMR spectroscopy. Using chemical shift, coupling constant and DMSO titration data it is found that these compounds adopt a C2-symmetric turn conformation featuring two intramolecular hydrogen bonds.     

Application of stir bar sorptive extraction for wine analysis

Stir bar sorptive extraction (SBSE) coupled with gas chromatography/mass spectrometry (GC/MS) was used to analyse wine samples for three applications: flavour and compositional analysis; 2,4,6-trichloroanisole (TCA), a common off-aroma in wine; and agrochemicals. SBSE was found to be orders of magnitude more sensitive than modern conventional methodology, allowing for lower detection and quantitation levels, and improved confirmation of identity; SBSE often gave better signal to noise in scan mode than other methods in selective ion monitoring (SIM) mode. With the help of their characteristic mass spectra all agrochemicals could be identified unambiguously at concentrations of 10 microg L(-1) in wine and a further 100 constituents were detected in a Cabernet Sauvignon sample. Thus it is now possible to analyse complex samples such as wine by scan mode, with better confirmation of identity, and without sacrificing sensitivity, where previously SIM methodology had to be used.     

Tobacco protein separation by aqueous two-phase extraction

Tobacco has long been considered as a host to produce large quantity of high-valued recombinant proteins. However, dealing with large quantities of biomass is a challenge for downstream processing. Aqueous two-phase extraction (ATPE) has been widely used in purifying proteins from various sources. It is a protein-friendly process and can be scaled up easily. In this paper, ATPE was studied for its applicability to recombinant protein purification from tobacco with egg white lysozyme as the model protein. Separate experiments with poly(ethylene glycol) (PEG)-salt-tobacco extract and PEG-salt-lysozyme were carried out to determine the partition behavior of tobacco protein and lysozyme, respectively. Two-level fractional factorial designs were used to study the effects of factors such as, PEG molecular mass, PEG concentration, the concentration of phase forming salt, sodium chloride concentration and pH, on protein partitioning. The results showed that, among the studied systems, PEG-sodium sulfate system was most suitable for lysozyme purification. Detailed experiments were conducted by spiking lysozyme into the tobacco extract. The conditions with highest selectivity of lysozyme over native tobacco protein were determined using a response surface design. The purification factor was further improved by decreasing the phase ratio along the tie line corresponding to the phase compositions with the highest selectivity. Under selected conditions the lysozyme yield was predicted to be 87% with a purification factor of 4 and concentration factor of 14. From this study, ATPE was shown to be suitable for initial protein recovery and partial purification from transgenic tobacco.     

Observation of a quadrupole plasmon mode for a colloidal solution of gold nanoprisms

The synthesis and optical properties of single crystalline gold nanoprisms have been investigated. A three-step mediated seed growth process in an aqueous solution generated gold nanoprisms with a relatively homogeneous size distribution. The purity of these nanostructures has allowed us to observe a weak quadrupole resonance in addition to a strong dipole resonance associated with these novel structures. The experimental optical spectra agree with discrete dipole approximation calculations that have been modeled from the dimensions of gold nanoprisms produced in this synthesis.     

The use of novel water-in-oil microemulsions in microemulsion electrokinetic chromatography

We describe the novel use of water-in-oil (W/O) microemulsions to achieve unique separations in microemulsion electrokinetic chromatography (MEEKC). The choice and concentration of the buffer type, surfactant and co-surfactant were all examined and optimized. Separations of a range of neutral and acidic analytes was shown to be markedly different to that obtained by (oil-in-water) O/W MEEKC. Neutral solutes are separated by virtue of their solubility (log P) values in O/W MEEKC with the more water-insoluble solutes migrating last. This separation process does not occur in W/O, as neutral solutes are not separated in order of log P.     

PHOTOSENSITIZATION WITH BACTERIOCHLORINS

Abstract— Biophysical and photobiological properties of a group of bacteriochlorins were compared with efficacy of these products for photodynamic therapy of murine tumors. Predictive factors for selective photosensitization in vivo include affinity binding to lipoproteins greater than albumin, extinction coefficient at the wavelength of irradiation and tumor/skin distribution. Efficacy was correlated with circulating plasma levels of the different sensitizers but not with the photodynamic therapy response in cell culture.     

Structural characterisation of a sol-gel copolymer synthesised from aliphatic and aromatic alkoxysilanes using 29Si-NMR spectroscopy

A hybrid organic-inorganic material was prepared from the hydrolytic co-condensation of 3-trimethoxysilylpropylmethacrylate (MAPTMS) and diphenyldiethoxysilane (DPHDES). The synthesis was performed in three steps: (1) MAPTMS pre-hydrolysis, (2) addition of DPHDES, and (3) further hydrolysis of the obtained mixture. ²⁹Si nuclear magnetic resonance spectroscopy was used to characterise the structural evolution of the mineral network during the synthesis. It revealed the catalytic effect of DPHDES on the condensation reaction of silanol groups, initially observed for pre-hydrolysed MAPTMS. In addition, it was clearly demonstrated that the formation of the highest condensed species of the aromatic oligomers (D₂ species) occurred only in the presence of aliphatic oligomers, which decrease the sterical hindrance about the silicon nuclei.