The production and characterisation of an antibody to detect the coccidiostat toltrazuril and its metabolite ponazuril

The production of an antibody to detect toltrazuril or its metabolite ponazuril is complicated due to structural constraints of conjugating these coccidiostats to a carrier protein. Therefore a search was carried out for a compound that shared a common substructure to use as an antigen mimic. The chosen compound, trifluoraminoether, was conjugated to two carrier proteins (HSA and BTG) and used in the immunisation of six rabbits. Two immunogen doses (1 mg and 0.1 mg) were also used. All six rabbits produced an immunological response to the hapten regardless of the carrier protein or immunogen dose used. The most sensitive polyclonal antibody produced, designated R609, was subsequently characterised. This antiserum exhibited an IC50 of 18 ng ml(-1) using a competitive ELISA format. Cross reactivity studies show that this serum is specific for toltrazuril and its metabolites (toltrazuril sulfoxide and toltrazuril sulfone) but does not cross-react with other coccidiostats such as halofuginone, nitroimidazoles or nicarbazin. This is the first reported production of an antibody capable of specifically binding toltrazuril and ponazuril.     

Michael addition of amines and thiols to dehydroalanine amides: a remarkable rate acceleration in water

In water, the rate of Michael addition of amines and thiols to dehydroalanine amides was greatly accelerated, leading to shorter reaction times and higher yields. The scope of the new conditions was tested with a range of amines, thiols, and dehydroalanine amides. The ease and efficiency of this method provides an attractive route to the synthesis of natural and unnatural amino acid derivatives.     

Stepped gradients on polymeric monolithic columns by photoinitiated grafting

The surface of polymethacrylate monoliths was functionalized by a post‐polymerization modification, by means of a novel photo‐initiated graft procedure where the charged monomer, sulfopropyl methacrylate, was controllably grafted stepwise, i.e. with incremental graft energies. The grafting approach was optimized using scanning capacitively coupled contactless conductivity detection. The effect of the localized ion exchange capacity and resultant gradient stationary phase upon ion‐exchange chromatographic retention, selectivity, and performance was investigated, and compared to a homogeneously grafted (isotropic) column. The gradient column provided reduced peak widths at half height for both cationic analytes, with a reduction of 34 and 33%, respectively, when compared to the isotropic column.     

Anti-plasmodial activity of some constituents of the root bark of Harungana madagascariensis LAM. (Hypericaceae)

Bazouanthrone (1), a new anthrone derivative, has been isolated from the root bark of Harungana madagascariensis, together with known compounds, feruginin A (2), harunganin (3), harunganol A (4), harunganol B (5), friedelan-3-one (6) and betulinic acid (7). The structure of the compound (1) was assigned as 3,5,8,9-tetrahydroxy-2,4,4-tri-(3,3-dimethylallyl)-6-methyl-1-(4H)-anthracenone, by means of spectroscopic analysis. The anti-plasmodial activity of the isolated compounds was evaluated in culture against W2 strain of Plasmodium falciparum. All the compounds were found to be active against the Plasmodium parasites with bazouanthrone (1) showing particular potency (IC50=1.80 microM).     

Triterpenoids

This review covers the isolation and structure determination of triterpenoids including squalene derivatives, lanostanes, cycloartanes, dammaranes, euphanes, tirucallanes, tetranortriterpenoids, quassinoids, lupanes, oleananes, friedelanes, uranes, hopanes, isomalabaricanes and saponins. The literature from January to December 2004 is reviewed and 243 references are cited.     

Long-Term Horizontal Vocal Directivity of Opera Singers: Effects of Singing Projection and Acoustic Environment

Vocal directivity refers to how directional the sound is that comes from a singer's mouth, that is, whether the sound is focused into a narrow stream of sound projecting in front of the singers or whether it is spread out all around the singer. This study investigates the long-term vocal directivity and acoustic power of professional opera singers and how these vary among subjects, among singing projections, and among vastly different acoustic environments. The vocal sound of eight professional opera singers (six females and two males) was measured in anechoic and reverberant rooms and in a recital hall. Subjects sang in four different ways: (1) paying great attention to intonation; (2) singing as in performance, with all the emotional connection intended by the composer; (3) imagining a large auditorium; and (4) imagining a small theatre. The same song was sung by all singers in all conditions. A head and torso simulator (HATS), radiating sound from its mouth, was used for comparison in all situations. Results show that individual singers have quite consistent long-term average directivity, even across conditions. Directivity varies substantially among singers. Singers are more directional than the standard HATS (which is a physical model of a talking person). The singer's formant region of the spectrum exhibits greater directivity than the lower-frequency range, and results indicate that singers control directivity (at least, incidentally) for different singing conditions as they adjust the spectral emphasis of their voices through their formants.     

Production of novel polymer monolithic columns, with stationary phase gradients, using cyclic olefin co-polymer (COC) optical filters

Polymer monolithic columns with controlled surface ligand density, providing stationary phase gradients within monolithic capillary columns, have been developed using photo-grafting through optical filters. Utilising commercially available cyclic olefin co-polymer (COC) films, the production of an optical filter capable of attenuating UV irradiation, in a tailored manner, was investigated. This novel optical filter was successfully applied to the surface modification of poly(BuMA-co-EDMA) monolithic columns in a multi-step grafting procedure. Fabricated columns were subjected to scanning capacitively coupled contactless conductivity (sC(4)D), to determine the distribution of the grafted functional groups, axially along the column. Further modification to produce a chelating stationary phase gradient of iminodiacetic acid (IDA) was demonstrated. To demonstrate the distribution of the IDA sites, a metal cation (Cu(2+)) was complexed to the IDA forming a chelate. Upon the formation of a complex of IDA with Cu(2+), an overall drop in conductive response was observed. The COC optical filter was also used in the fabrication of a grafted gradient of strong cation exchanger (SCX), sulphopropyl methacrylate (SPM) upon a polymer monolith, demonstrating the broader applicability of such a filter.