An allelic series reveals essential roles for FY in plant development in addition to flowering-time control

The autonomous pathway functions to promote flowering in Arabidopsis by limiting the accumulation of the floral repressor FLOWERING LOCUS C (FLC). Within this pathway FCA is a plant-specific, nuclear RNA-binding protein, which interacts with FY, a highly conserved eukaryotic polyadenylation factor. FCA and FY function to control polyadenylation site choice during processing of the FCA transcript. Null mutations in the yeast FY homologue Pfs2p are lethal. This raises the question as to whether these essential RNA processing functions are conserved in plants. Characterisation of an allelic series of fy mutations reveals that null alleles are embryo lethal. Furthermore, silencing of FY, but not FCA, is deleterious to growth in Nicotiana. The late-flowering fy alleles are hypomorphic and indicate a requirement for both intact FY WD repeats and the C-terminal domain in repression of FLC. The FY C-terminal domain binds FCA and in vitro assays demonstrate a requirement for both C-terminal FY-PPLPP repeats during this interaction. The expression domain of FY supports its roles in essential and flowering-time functions. Hence, FY may mediate both regulated and constitutive RNA 3'-end processing.

Growth setbacks in new firms

New firms in emerging industries are subject to complex dynamic processes which defy the attempts at prediction embodied in business conjectures. Discontinuous growth is common, but the issue of interruptions in the early growth of new firms has not been adequately addressed in the mainstream literature. We examine the prevalence of interruptions to growth in a cohort study of the growth trajectories of firms founded in 1990, then look to cases studies of individual firms to investigate underlying causes. We find that substantial growth is rare and continuous growth unusual, and that growth interruptions are the result of both internal and external dynamics. The managers of growing firms face shortages of vital resources and significant problems of resource synchronisation and coordination, many of which can lead to what are, in effect, changes of phase state. Meanwhile, the volatile environment of an emerging industry presents particular problems to young firms which have not yet built up reserves to sustain them through short-term crises. However, problem solving by those that survive provides an important source of learning which can underpin their future development. © 2004 Elsevier Ltd. All rights reserved.

Copper homeostasis in Salmonella is atypical and copper-CueP is a major periplasmic metal complex.

Salmonella enterica sv. typhimurium (S. enterica sv. Typhimurium) has two metal-transporting P(1)-type ATPases whose actions largely overlap with respect to growth in elevated copper. Mutants lacking both ATPases over-accumulate copper relative to wild-type or either single mutant. Such duplication of ATPases is unusual in bacterial copper tolerance. Both ATPases are under the control of MerR family metal-responsive transcriptional activators. Analyses of periplasmic copper complexes identified copper-CueP as one of the predominant metal pools. Expression of cueP was recently shown to be controlled by the same metal-responsive activator as one of the P(1)-type ATPase genes (copA), and copper-CueP is a further atypical feature of copper homeostasis in S. enterica sv. Typhimurium. Elevated copper is detected by a reporter construct driven by the promoter of copA in wild-type S. enterica sv. Typhimurium during infection of macrophages. Double mutants missing both ATPases also show reduced survival inside cultured macrophages. It is hypothesized that elevated copper within macrophages may have selected for specialized copper-resistance systems in pathogenic microorganism such as S. enterica sv. Typhimurium.

Microfabrication and application of reservoir pins for liquid transfer in biotechnology

Microarraying involves laying down genetic elements onto a solid substrate for DNA analysis on a massively parallel scale. Microarrays are prepared using a pin-based robotic platform to transfer liquid samples from microtitre plates to an array pattern of dots of different liquids on the surface of glass slides where they dry to form spots diameter < 200 μm. This paper presents the design, materials selection, micromachining technology and performance of reservoir pins for microarraying. A conical pin is produced by (i) conventional machining of stainless steel or wet etching of tungsten wire, followed by (ii) micromachining with a focused laser to produce a microreservoir and a capillary channel structure leading from the tip. The pin has a flat end diameter < 100 μm from which a 500 μm long capillary channel < 15 μm wide leads up the pin to a reservoir. Scanning electron micrographs of the metal surface show roughness on the scale of 10 μm, but the pins nevertheless give consistent and reproducible spotting performance. The pin capacity is 80 nanolitres of fluid containing DNA, and at least 50 spots can be printed before replenishing the reservoir. A typical robot holds can hold up to 64 pins. This paper discusses the fabrication technology, the performance and spotting uniformity for reservoir pins, the possible limits to miniaturization of pins using this approach, and the future prospects for contact and non-contact arraying technology.

Jet diameters and velocity profiles in continuous ink-jets

Theoretical predictions of the diameters of continuous ink-jets downstream of long nozzles are generalized to include the important cases of ink-jet fluids and shorter nozzles where the velocity profile at the nozzle exit is undeveloped (non-parabolic). Comparisons of the new predictions with experiments and simulations are made for fairly long nozzles with tapered profiles and short nozzles with conical profiles; experimental and simulated profiles are also compared downstream of the nozzle exit for both industrial and large scale ink-jet print heads. Precise measurements of the un-modulated jet diameters downstream of the nozzle exit can set really useful limits to the possible shapes of the flow profile right at the nozzle exit, and in particular allow some assessment of the axial velocity gradients and fluid shear rates at the nozzle exit where direct speed measurement is usually impractical. Simulations allow further study of the relaxation of the velocity profile downstream of the nozzle exit, and are reported for both un-modulated and modulated CIJ jetting. Implications of this work include speeding up CIJ simulations, absolute calibration of the applied CIJ system modulation, and the likely magnitude of dynamic surface tension effects on observed CIJ satellite speeds.

On Mixing and Shaking : Structure and Dynamics of Turbulent Stratified Flames

The drive for low emission combustion systems encourages applications using premixed flames. Yet in many applications, considerations of flame stability or mixing times lead to systems with neither premixed nor diffusion flames, which are often called technically premixed or stratified flames. In this talk we discuss the current state of understanding of the effect of mixing and extent of stratification on the structure, microstructure and dynamics of selected turbulent stratified flames. Over the past few years, a significant database of scalar and velocity data has been built to analyze the effects of unmixedness on local and global flame structure. Microscale studies of the flame structures show in detail how the effect of local stratification affects (or not!) the flame structure, flame surface density and scalar dissipation rates, and production of selected species. The experiments place exacting demands on current spectroscopic diagnostics, and reveal the progress and limits to our understanding of turbulent flames in general. The dynamics of stratified flames with respect to instabilities is also shown to be very rich, as the particular shape of the flames and the stabilization points are is significantly affected by the fuel distribution, modifying the rate and location of heat release, and thus the coupling with the surrounding acoustics and determining the onset of self-excitations.