Resolving genetic functions within microbial populations: In situ analyses using rRNA and mRNA stable isotope probing coupled with single-cell raman-fluorescence in situ hybridization

Prokaryotes represent one-half of the living biomass on Earth, with the vast majority remaining elusive to culture and study within the laboratory. As a result, we lack a basic understanding of the functions that many species perform in the natural world. To address this issue, we developed complementary population and single-cell stable isotope (C-13)-linked analyses to determine microbial identity and function in situ. We demonstrated that the use of rRNA/mRNA stable isotope probing (SIP) recovered the key phylogenetic and functional RNAs. This was followed by single-cell physiological analyses of these populations to determine and quantify in situ functions within an aerobic naphthalene-degrading groundwater microbial community. Using these culture-independent approaches, we identified three prokaryote species capable of naphthalene biodegradation within the groundwater system: two taxa were isolated in the laboratory (Pseudomonas fluorescens and Pseudomonas putida), whereas the third eluded culture (an Acidovorax sp.). Using parallel population and single-cell stable isotope technologies, we were able to identify an unculturable Acidovorax sp. which played the key role in naphthalene biodegradation in situ, rather than the culturable naphthalene-biodegrading Pseudomonas sp. isolated from the same groundwater. The Pseudomonas isolates actively degraded naphthalene only at naphthalene concentrations higher than 30 mu M. This study demonstrated that unculturable microorganisms could play important roles in biodegradation in the ecosystem. It also showed that the combined RNA SIP-Raman-fluorescence in situ hybridization approach may be a significant tool in resolving ecology, functionality, and niche specialization within the unculturable fraction of organisms residing in the natural environment.

Infinite Dimensional Banach spaces of functions with nonlinear properties

The aim of this paper is to show that there exist infinite dimensional Banach spaces of functions that, except for 0, satisfy properties that apparently should be destroyed by the linear combination of two of them. Three of these spaces are: a Banach space of differentiable functions on Rn failing the Denjoy-Clarkson property; a Banach space of non Riemann integrable bounded functions, but with antiderivative at each point of an interval; a Banach space of infinitely differentiable functions that vanish at infinity and are not the Fourier transform of any Lebesgue integrable function.

Inline Interdigital Pseudo-Elliptic Helical Resonator Filters

A new inline coupling topology for narrowband helical resonator filters is proposed that allows to introduce selectively located transmission zeros (TZs) in the stopband. We show that a pair of helical resonators arranged in an interdigital configuration can realize a large range of in-band coupling coefficient values and also selectively position a TZ in the stopband. The proposed technique dispenses the need for auxiliary elements, so that the size, complexity, power handling and insertion loss of the filter are not compromised. A second order prototype filter with dimensions of the order of 0.05 lambda, power handling capability up to 90 W, measured insertion loss of 0.18 dB and improved selectivity is presented.

Nematode neuropeptides: Localization, isolation and functions

Historically, peptidergic substances (in the form of neurosecretions) were linked to moulting in nematodes. More recently, there has been a renewal of interest in nematode neurobiology, initially triggered by studies demonstrating the localization of peptide immunoreactivities to the nervous system. Here, David Brownlee, Ian Fairweather, Lindy Holden-Dye and Robert Walker will review progress on the isolation of nematode neuropeptides and efforts to unravel their physiological actions and inactivation mechanisms. Future avenues for research are suggested and the potential exploitation of peptidergic pathways in future therapeutic strategies highlighted.