New approaches to the synthesis of antiviral nucleosides.

There is a pressing, worldwide need for new antiviral agents. The chemical synthesis of novel nucleosides for chemotherapeutic screening usually involves multistage processes which can be time consuming. The application of enzymatic methods for the synthesis and modification of antiviral nucleosides shows great promise because of the simplicity and high specificity of enzymatic reactions.     

New approaches to enzymatic glycoside synthesis through directed evolution

The expanding field of glycobiology requires tools for the synthesis of structurally defined oligosaccharides and glycoconjugates, while any potential therapeutic applications of sugar-based derivates would require access to substantial quantities of such compounds. Classical chemical approaches are not well suited for such large-scale syntheses, thus enzymatic approaches are sought. Traditional routes to the enzymatic assembly of oligosaccharides have involved the use of either Nature’s own biosynthetic enzymes, the glycosyl transferases, or glycosidases run in transglycosylation mode. However, each approach has drawbacks that have limited its application. Glycosynthases are mutant glycosidases in which the catalytic nucleophile has been replaced by mutation, inactivating them as hydrolases. When used in conjunction with glycosyl fluorides of the opposite anomeric configuration to that of the substrate, these enzymes function as highly efficient transferases, frequently giving stoichiometric yields of products. Further improvements can be obtained through directed evolution of the gene encoding the enzyme in question, but this requires the ability to screen very large libraries of catalysts. In this review we survey new screening methods for the formation of glycosidic linkages using high-throughput techniques, such as FACS, chemical complementation, and robot-assisted ELISA assays. Enzymes were evolved to have higher catalytic activity with their natural substrates, to show altered substrate specificities or to be promiscuous for efficient application in oligosaccharide, glycolipid, and glycoprotein synthesis.     

New approaches to the chemical synthesis of bioactive oligosaccharides.

The past year has seen some major advances in the area of carbohydrate synthesis using chemical methods. Progress in all areas of synthetic methodology, including new protecting groups and coupling methods, has been reported. A number of complex carbohydrate structures have been prepared using known, as well as new, methods. The goal to allow nonspecialists access to defined carbohydrate structures for biochemical, biophysical and biological studies has drawn closer by the introduction of two approaches towards synthesis automation. A one-pot glycosylation strategy utilized computer-assisted synthesis planning and the first solid-phase automated synthesizer was introduced very recently.     

New approaches towards automated oligoribonucleotide synthesis.

Several protecting group combinations have been investigated to achieve an automated oligoribonucleotide synthesis on a solid support. The use of 2'-O-tert-butyldimethylsilyl-5'-O-monomethoxytrityl-ribonucleoside -3'-(p-nitrophenylethyl, N,N-diethyl)-phosphoramidites in conjunction with beta-eliminating blocking groups at the aglycone gave satisfactory results. An inverse protecting group strategy based upon analogously substituted 5'-O-2-dansylethoxycarbonyl-2'-O-4- methoxytetrahydropyran-4-yl-phosphoramidites can be regarded as another promising approach.     

New approaches to antibiotic discovery

New antibiotics are urgently required by human medicine as pathogens emerge with developed resistance to almost all antibiotic classes. Pioneering approaches, methodologies and technologies have facilitated a new era in antimicrobial discovery. Innovative culturing techniques such as iChip and co-culturing methods which use ‘helper’ strains to produce bioactive molecules have had notable success. Exploiting antibiotic resistance to identify antibacterial producers performed in tandem with diagnostic PCR based identification approaches has identified novel candidates. Employing powerful metagenomic mining and metabolomic tools has identified the antibiotic’ome, highlighting new antibiotics from underexplored environments and silent gene clusters enabling researchers to mine for scaffolds with both a novel mechanism of action and also few clinically established resistance determinants. Modern biotechnological approaches are delivering but will require support from government initiatives together with changes in regulation to pave the way for valuable, efficacious, highly targeted, pathogen specific antimicrobial therapies.     

New approaches to high-throughput phasing

Recent progress in macromolecular phasing, in part stimulated by the high-throughput structural biology initiatives, has made this crucial stage of the elucidation of crystal structures easier and more automatic. A quick soak in various salts leads to the rapid incorporation of the anomalously scattering ions, suitable for phasing by MAD (multiwavelength anomalous dispersion), SAD (single-wavelength anomalous dispersion) or MIR (multiple isomorphous replacement) methods. The availability of stable synchrotron beam lines equipped with elaborate hardware control and sophisticated data processing programs makes it possible to collect very accurate diffraction data and to solve structures from the very weak anomalous signal of such atoms as sulfur or phosphorus, inherently present in macromolecules. The current progress in phasing, coupled with the parallel advances in protein crystallization, diffraction data collection and so on, suggests that, in the near future, the process of macromolecular crystal structure elucidation may become fully automatic.     

New approaches to transillumination imaging

A review of the current state of transillumination imaging for the detection and diagnosis of breast cancer and the difficulties that impede more widespread acceptance of the methods is presented. An outline is given of the physical models that may be used to describe the propagation and scattering of light in a tissue matrix and how these models might be valuable in identifying imaging improvements. Some of the proposals for future imaging arrangements are described and the preliminary work on a system for light transmission computed tomography is presented.     

New approaches to measuring nausea

Valid measures of nausea are needed to evaluate the various treatments used to counter the nausea produced by chemotherapy. The overall nausea intensity (ONI) produced by 17 chemotherapy drugs was estimated by 17 physicians and 8 nurses, and 25 patients undergoing chemotherapy described the subjective qualities and ONI of their nausea on a modified form of the McGill Pain Questionnaire. The scores for the affective and miscellaneous categories of words in the questionnaire were found to correlate significantly with the physicians'' and nurses'' ONI estimates. The results formed the basis for the Nausea Questionnaire, which provided three indices of nausea: a nausea rating index (NRI), ONI and intensity of nausea according to a visual analogue scale (VAS). All three indices correlated significantly with the physicians'' and nurses'' ONI estimates and were significantly intercorrelated. All three also provided significant differences when the scores of patients who had received cisplatin or 5-fluorouracil were compared. The results indicate that the Nausea Questionnaire provides three valid indices of the subjective experience of nausea.     

A novel,stereospecific synthesis of β-D-glucopyranosyl phosphate

In alkaline solution, periodate ions form complexes with three consecutive hydroxyl groups in an axial-equatorial-axial arrangement but not with three syn-axial hydroxyl groups. Borate ions form complexes with three syn-axial hydroxyl groups but not with the axeqax sequence. Accordingly, cis-inositol gives a 1,2,3-periodate and a 1,3,5-borate complex. Cations form complexes with either type of conformation, but complex formation provides little energy towards achieving it by ring inversion. Lanthanide ions have been used to establish the sites of complex formation.