ALBERT

Notes: TlpC is encoded in the second chemotaxis operon of Rhodobacter sphaeroides. This protein shows some homology to membrane-spanning chemoreceptors of many bacterial species but, unlike these, is essential for R. sphaeroides chemotaxis to all compounds tested. Genomic replacement of tlpC with a C-terminal gfp fusion demonstrated that TlpC localized to a discrete cluster within the cytoplasm. Immunogold electron microscopy also showed that TlpC localized to a cytoplasmic electron-dense region. Correct TlpC–GFP localization depended on the downstream signalling proteins, CheW3, CheW4 and CheA2, and was tightly linked to cell division. Newly divided cells contained a single cluster but, as the cell cycle progressed, a second cluster appeared close to the initial cluster. As elongation continued, these clusters moved apart so that, on septation, each daughter cell contained a single TlpC cluster. The data presented suggest that TlpC is either a cytoplasmic chemoreceptor responding to or integrating global signals of metabolic state or a novel and essential component of the chemotaxis signalling pathway. These data also suggest that clustering is essential for signalling and that a mechanism may exist for targeting and localizing proteins within the bacterial cytoplasm.

Notes: The purple photosynthetic bacterium Rhodobacter sphaeroides has three loci encoding multiple homologues of the bacterial chemosensory proteins: 13 putative chemoreceptors, four CheW, four CheA, six CheY, two CheB and three CheR. Previously, studies have shown that, although deletion of cheOp1 led to only minor changes in behaviour, deletion of cheOp2 led to a loss of taxis. The third locus encodes two CheA, one CheR, one CheB, one CheW, one CheY, a putative cytoplasmic chemoreceptor (TlpT) and a protein showing homology to the chromosomal partitioning factor Soj (designated Slp). Here, we show that every protein encoded by this locus is essential for normal chemotaxis. Phototaxis is also dependent upon all the components of this locus, except CheB2 and Slp. The two putative CheA proteins encoded in this locus are unusual. CheA3 has only the P1 domain and the P5 regulatory domain linked by a large internal domain, whereas CheA4 lacks the P1 and P2 domains required for phosphorylation and response regulator binding. These data indicate that the minimal set of proteins required for normal chemotaxis in R. sphaeroides is all the proteins encoded by cheOp2 and the third chemotaxis locus, and that the multiple chemosensory protein homologues found in R. sphaeroides are not redundant.

Notes: Antibody phage display libraries (Griffin and Tomlinson I) displaying antibody genes and maintained and amplified in Escherichia coli were used to isolate antibodies to the hapten target microcystin LR (1000 Da) conjugated to either bovine serum albumin or keyhole limpet haemocyanin. In competition enzyme-linked immunosorbent assay, bacterially expressed antibodies selected via the Griffin library showed at least 300 times greater sensitivity than those isolated from the Tomlinson library, for free microcystin. Bacterially expressed phage antibody libraries provide a rapid and relatively easy route for the selection of monoclonal antibodies specific for even the most difficult of antigenic targets such as free haptens.

Notes: There is growing concern about the fate and toxicity of herbicides to non-target receptors and an increasing need to measure these analytes sensitively. The responses of cellular and immunological biosensors to four commonly used herbicides (atrazine, diuron, mecoprop and paraquat) were investigated. In combination, these sensors assess toxicity and quantify concentrations of herbicides present in extracts from soil. The bioluminescence response of the lux-marked bacterial biosensor Escherichia coli HB101 was determined in aqueous extracts from soil to indicate toxicity. Smaller concentrations caused a toxic response for all four herbicides recovered from the Insch series than for those recovered from spiked water samples, but this was not a result of biodegradation of herbicides in the soil. This suggests that intrinsic soil factors may be altering the bioavailable fraction of herbicides, making them more toxic than equivalent concentrations in water.Herbicide concentrations were determined using immunological biosensors consisting of stabilized recombinant single chain antibodies (stAbs) specific for the four different groups of herbicides. These stAb fragments retain functionality in organic solvents such as methanol commonly used in soil extraction. Anti-atrazine, mecoprop, diuron and paraquat stAbs were successfully used to identify and quantify herbicides present in aqueous and methanol extracts from soil. The amounts recovered from immunoassay analysis were compared with chemical analysis using high performance liquid chromatography, and the two methods correlated. These stAb fragments might provide a more rapid and sensitive means of quantifying trace amounts of herbicides and their metabolites in aqueous and methanol extracts from soil.

Notes: Abstract An experiment was carried out over 2 years to evaluate the effects of increasing the proportion of cereal-based concentrates in diets containing high-digestibility and conventional medium-digestibility grass silages on the dry-matter (DM) intake, liveweight gain and carcass composition of beef cattle, and to examine the effects of grazed grass and the ratio of grass silage:concentrates in the diet on the fatty acid composition of selected muscle tissues. Late-maturing steers (n = 231) were offered diets based on high-digestibility (HD) (0·743 digestible organic matter (DOM) in DM) or medium-digestibility (MD) (0·643 DOM in DM) grass silages supplemented with barley/soyabean meal-based concentrates. The concentrates constituted 0·20, 0·40, 0·60 and 0·80 of total DM of the diets, which were offered ad libitum (AL). The two diets, which contained 0·80 concentrates, were also offered at 0·80 of AL intake. A further group of fourteen animals were given the medium-digestibility silage only for 5 months and then grazed perennial ryegrass pastures for a further 5 months (silage/pasture treatment). For the diets containing HD silage and 0·20, 0·40, 0·60 and 0·80 concentrate, and 0·80 concentrate at 0·8 of AL intake, the DM intakes were 9·4, 10·2, 10·4, 10·2 and 8·1 (s.e. 0·16) kg d−1, respectively, and daily carcass gains were 0·67, 0·78, 0·77, 0·79 and 0·62 (s.e. 0·029) kg d−1, respectively; for those containing MD silage and 0·20, 0·40, 0·60 and 0·80 concentrate, and 0·80 concentrate at 0·8 of AL, the DM intakes were 8·2, 9·3, 10·1, 10·1 and 8·0 (s.e. 0·16) kg d−1, respectively, and daily carcass gains were 0·38, 0·48, 0·64, 0·77 and 0·56 (s.e. 0·029) kg d−1 respectively. Increasing the proportion of concentrates in silage-based diets decreased the concentration of omega-3 (ω-3) polyunsaturated fatty acids (PUFA) (P 〈 0·001) and increased the concentration of ω-6 PUFA (P 〈 0·001) in muscle. Cattle on the silage/pasture treatment had the highest concentration of ω-3 PUFA in muscle (51 g kg−1 lipid), this value being over three times that for animals given diets containing MD silage and 0·80 concentrate in the diet. These results demonstrate the potential of HD silage made from perennial ryegrass relative to high concentrate diets. The consumption of pasture-finished beef could make a significant contribution towards increasing the intake of ω-3 PUFA in the human diet.

Notes: The Z-pinch-driven hohlraum (ZPDH) [J. H. Hammer et al., Phys. Plasmas 6, 2129 (1999)] is a promising approach to high yield inertial confinement fusion currently being characterized in experiments on the Sandia Z accelerator [M. E. Cuneo et al., Phys. Plasmas 8, 2257 (2001)]. Simulations show that capsule radiation symmetry, a critical issue in ZPDH design, is governed primarily by hohlraum geometry, dual-pinch power balance, and pinch timing. In initial symmetry studies on Z without the benefit of a laser backlighter, highly-asymmetric pole-hot and equator-hot single Z-pinch hohlraum geometries were diagnosed using solid low density foam burnthrough spheres. These experiments demonstrated effective geometric control and prediction of polar flux symmetry at the level where details of the Z-pinch implosion and other higher order effects are not critical. Radiation flux symmetry achieved in Z double-pinch hohlraum configurations exceeds the measurement sensitivity of this self-backlit foam ball symmetry diagnostic. To diagnose radiation symmetry at the 2%–5% level attainable with present ZPDH designs, high-energy x rays produced by the recently-completed Z-Beamlet laser backlighter are being used for point-projection imaging of thin-wall implosion and symmetry capsules. © 2002 American Institute of Physics.

Notes: Fractures from tests on 2014-T6511 and 2024-T3 test coupons under specially designed programmed loading reveal voids with distinct fatigue markings. These ‘fatigue voids’ appear to form as a consequence of the separation of noncoherent secondary particulates from the matrix in early fatigue. The process of their formation is through the initiation, growth and coalescence of multiple interfacial cracks around the particulate. Such voids become visible on the fatigue fracture surface if and when the crack front advances through them. In vacuum, each fatigue void is the potential initiator of an embedded penny-shaped crack. The one closest to the specimen surface is likely to become the dominant crack, indicating that fatigue voids appear to be the likely origins of the dominant crack in vacuum. In air, the dominant crack forms at the notch surface and grows much faster, giving less opportunity for multiple internal cracks to spawn off from the innumerable internal fatigue-voids. Thus in air, fatigue voids do not appear to affect the fatigue process at low and intermediate growth rates. At high crack growth rates involving considerable crack tip shear, slip planes with particulate concentration offer the path of least resistance. This explains the increasing density of fatigue voids with growth rate. Very high growth rates signal the onset of a quasi-static crack growth component that manifests itself through growing clusters of microvoid coalescence associated with static fracture. Fatigue voids are likely to form in other Al-alloys with secondary noncoherent particulates. They have nothing in common with microvoids associated with ductile fracture.

Notes: There is increasing public, governmental and commercial interest in the welfare of intensively farmed fish and stocking density has been highlighted as an area of particular concern. Here we draw scientific attention and debate to this emerging research field by reviewing the evidence for effects of density on rainbow trout. Although no explicit reference to ‘welfare’ has been made, there are 43 studies which have examined the effects of density on production and physiological parameters of rainbow trout. Increasing stocking density does not appear to cause prolonged crowding stress in rainbow trout. However, commonly reported effects of increasing density are reductions in food conversion efficiency, nutritional condition and growth, and an increase in fin erosion. Such changes are indicative of a reduced welfare status—although the magnitude of the effects has tended to be dependent upon study-specific conditions. Systematic observations on large scale commercial farms are therefore required, rather than extrapolation of these mainly small-scale experimental findings. There is dispute as to the cause of the observed effects of increasing density, with water quality deterioration and/or an increase in aggressive behaviour being variously proposed. Both causes can theoretically generate the observed effects of increasing density, and the relative contribution of the two causes may depend upon the specific conditions. However, documentation of the relationship between density and the effects of aggressive behaviour at relevant commercial densities is lacking. Consequently only inferential evidence exists that aggressive behaviour generates the observed effects of increasing density, whereas there is direct experimental evidence that water quality degradation is responsible. Nevertheless, there are contradictory recommendations in the literature for key water quality parameters to ensure adequate welfare status. The potential for welfare to be detrimentally affected by non-aggressive behavioural interactions (abrasion, collision, obstruction) and low densities (due to excessive aggressive behaviour and a poor feeding response) have been largely overlooked. Legislation directly limiting stocking density is likely to be unworkable, and a more practical option might be to prescribe acceptable levels of water quality, health, nutritional condition and behavioural indicators.

Notes: This is the second of two articles about the use of case studies in management training and development. In the first article the authors explained the potential benefits of using this method as well as some of the dangers. They also explained the methodology of running cases effectively. In this second article they draw on their extensive experience of writing and using case studies in the UK and abroad to deal with important further aspects. These are the skills involved in writing cases studies, their use in assessment and cross-cultural issues.