Differentiation of Micromonospora isolates from a coastal sediment in Wales on the basis of Fourier transform infrared spectroscopy, 16S rRNA sequence analysis, and the amplified fragment length polymorphism technique

A number of actinomycetes isolates were recovered from coastal sediments in Aberystwyth (Wales, United Kingdom) with standard isolation techniques. Most of them were putatively assigned to the genera Streptomyces and Micromonospora on the basis of their morphological characteristics, and there appeared to be no difference whether the isolation media contained distilled water or seawater. A group of 20 Micromonospora isolates was selected to undergo further polyphasic taxonomic investigation. Three approaches were used to analyze the diversity of these isolates, 16S rDNA sequencing, fluorescent amplified fragment length polymorphism (AFLP), and Fourier transform infrared spectroscopy (FT-IR). The 16S rDNA sequence analysis confirmed that all of these isolates should be classified to the genus Micromonospora, and they were analyzed with a group of other Micromonospora 16S rDNA sequences available from the Ribosomal Database Project. The relationships of the 20 isolates were observed after hierarchical clustering, and almost identical clusters were obtained with these three techniques. This has obvious implications for high-throughput screening for novel actinomycetes because FT-IR spectroscopy, which is a rapid and reliable whole-organism fingerprinting method, can be applied as a very useful dereplication tool to indicate which environmental isolates have been cultured previously.     

The use of chemical profiling for monitoring metabolic changes in artificial soil slurries caused by horizontal gene transfer

This study explores the utility of Fourier transform infra-red spectroscopy (FT-IR) as a metabolomic tool to detect changes in water-extractable chemical profile resulting from horizontal gene transfer (HGT) events in artificial soil slurries. A GFP–Km (Green fluorescent protein–kanamycin) cassette tagged HGT recipient Acinetobacter strain ADPWH67 with the salicylate hydroxylase gene (salA) disrupted was introduced to slurries containing either sterile or non-sterile soil. The subsequent addition of naked salA DNA allowed the specific monitoring of HGT events by enumerating GFP-expressing colonies on minimal media with salicylate as a sole carbon source. DNA sequencing confirmed that salA was restored in these transformants. Gene transformation frequencies of around 10⁻⁶ were achieved in the presence of sterile and non-sterile soils. Aqueous extracts of the soil slurries were then analyzed using FT-IR in order to ascertain whether any shifts in chemical profile could be detected. We found that following HGT events FT-IR chemical profiles were clearly altered when analyzed with multivariate statistics. Furthermore, these changes could be explained by differences in key chemical signatures including salicylate as well as other biomolecules found in soils. The slurry extracts were also subjected to GC-MS which confirmed the results of FT-IR analyses. FT-IR was therefore demonstrated to have utility for the rapid screening of metabolomic changes in soils following effective HGT events. In addition, this approach could potentially link specific metabolite changes with corresponding catabolic genes.Electronic Supplementary Material Supplementary material is available for this article at and is accessible for authorized users.     

Glycosylation of recombinant antibody therapeutics

The adaptive immune system has the capacity to produce antibodies with a virtually infinite repertoire of specificities. Recombinant antibodies specific for human targets are established in the clinic as therapeutics and represent a major new class of drug. Therapeutic efficacy depends on the formation of complexes with target molecules and subsequent activation of downstream biologic effector mechanisms that result in elimination of the target. The activation of effector mechanisms is dependent on structural characteristics of the antibody molecule that result from posttranslational modifications, in particular, glycosylation. The production of therapeutic antibody with a consistent human glycoform profile has been and remains a considerable challenge to the biopharmaceutical industry. Recent research has shown that individual glycoforms of antibody may provide optimal efficacy for selected outcomes. Thus a further challenge will be the production of a second generation of antibody therapeutics customized for their clinical indication.     

Cytokine medicines in clinical practice: current issues

Cytokine medicines have been licensed for the treatment of rheumatoid arthritis since 2000. The rheumatology community has accrued a large amount of experience in the use of these medications. This experience has led to the development of guidelines for their use that include ongoing vigilance for long term adverse events and efficacy using the Biologics Register. Delivery of these expensive therapies has prompted extensive system developments within rheumatology. The cytokine medicines have provided important tools to probe the pathogenesis of rheumatoid and other inflammatory diseases. Further cytokine medicines, in various stages of development, are on the horizon and continue to stimulate excitement within this fast expanding field.     

A series of bisaryl imidazolidin-2-ones has shown to be selective and orally active 5-HT2C receptor antagonists

Bisaryl cyclic ureas have been identified as high affinity 5-HT2C receptor antagonists with selectivity over 5-HT2A and 5-HT2B. Compounds such as 8 and 22 have shown oral activity in a centrally mediated pharmacodynamic model of 5-HT2C function in rodents.     

Genetic variation in a Pacific Island land snail: population history versus current drift and selection

Previous studies of Partula land snails from the Society Islands, French Polynesia, have shown that there can be striking differences in shell shape, colour and banding pattern between nearby populations, even in the absence of any obvious geographical barriers to the movement of snails, or environmental gradients. Elsewhere, there may be relative uniformity over large distances. Analysis of a mitochondrial gene from Partula taeniata (M?rch) shows a similar pattern. The relative frequencies of two mitochondrial haplotypes change abruptly over small distances, seemingly independent of the environment. Although the transition roughly coincides with clines in the frequencies of some morphological characteristics, it appears to be unrelated to others. It is likely that many of the differences accumulated while populations were isolated from one another, through the effects of random genetic drift and selection. Isolation of populations may have occurred as a result of demographic changes, or during the process of colonization if occasional long-distance migrants establish populations ahead of the main invading front. Current genetic drift, even without restrictions to gene flow, may contribute to genetic patchiness on a small scale, although it is likely that conspicuous characteristics such as shell colours and banding patterns are also influenced by selection.     

Serum antibodies to group A streptococcal extracellular and cell-associated antigens in Egyptians with post-streptococcal diseases

We investigated serum antibodies to a comprehensive array of group A streptococcal antigens and superantigens in Egyptian subjects. Antibodies to Streptococcus pyogenes cell-associated proteins and to proteins released by rapidly dividing S. pyogenes were compared in four patient groups with different post-streptococcal diseases and in healthy controls. Enzyme-linked immunosorbent assays showed that total Ig and IgG to extracellular antigens were significantly higher in patients with acute rheumatic fever (ARF) compared to healthy controls, but no differences were found in either total Ig or IgG titres to cell-associated proteins between any of the groups. Western blotting showed that multiple extracellular and cell-associated antigens, covering a wide range of molecular masses, were recognised by all sera, including healthy controls. No evidence was obtained for putative dominant antigens associated with any disease group, although a low molecular mass cell-associated protein (approximately 4 kDa) was clearly recognised by two-thirds of subjects irrespective of disease status. These findings demonstrate that raised serum Ig and IgG titres to extracellular, but not cell-associated, S. pyogenes antigens are a feature of ARF in this population, and suggest that multiple S. pyogenes antigens contribute to this response.     

3-(4-Piperidinyl)- and 3-(8-aza-bicyclo[3.2.1]oct-3-yl)-2-phenyl-1H-indoles as bioavailable h5-HT2A antagonists

A series of 3-(4-piperidinyl)- and 3-(8-aza-bicyclo[3.2.l]oct-3-yl)-2-phenyl-1H-indoles have been prepared and evaluated as ligands for the h5-HT_2A receptor. 3-(8-Phenethyl-8-aza-bicyclo[3.2.l]oct-3-yI)-2-phenyl-1H-indole is a high-affinity (1.2 nM), selective (>800 fold over h5-HT_2C and hD₂ receptors) antagonist at the h5-HT_2A receptor with oral bioavailability in rats.