Cysteine proteinase-1 and cut protein isoform control dendritic innervation of two distinct sensory fields by a single neuron.

Dendrites often exhibit structural changes in response to local inputs. Although mechanisms that pattern and maintain dendritic arbors are becoming clearer, processes regulating regrowth, during context-dependent plasticity or after injury, remain poorly understood. We found that a class of Drosophila sensory neurons, through complete pruning and regeneration, can elaborate two distinct dendritic trees, innervating independent sensory fields. An expression screen identified Cysteine proteinase-1 (Cp1) as a critical regulator of this process. Unlike known ecdysone effectors, Cp1-mutant ddaC neurons pruned larval dendrites normally but failed to regrow adult dendrites. Cp1 expression was upregulated/concentrated in the nucleus during metamorphosis, controlling production of a truncated Cut homeodomain transcription factor. This truncated Cut, but not the full-length protein, allowed Cp1-mutant ddaC neurons to regenerate higher-order adult dendrites. These results identify a molecular pathway needed for dendrite regrowth after pruning, which allows the same neuron to innervate distinct sensory fields.

Outer Membrane Vesicle Production in Escherichia coli Relieves Envelope Stress and is Modulated by Changes in Peptidoglycan

Bacterial outer membrane vesicles (OMVs) are spherical buds of the outer membrane (OM) containing periplasmic lumenal components. OMVs have been demonstrated to play a critical part in the transmission of virulence factors, immunologically active compounds, and bacterial survival, however vesiculation also appears to be a ubiquitous physiological process for Gram-negative bacteria. Despite their characterized biological roles, especially for pathogens, very little is known about their importance for the originating organism as well as regulation and mechanism of production. Only when we have established their biogenesis can we fully uncover their roles in pathogenesis and bacterial physiology. The overall goal of this research was to characterize bacterial mutants which display altered vesiculation phenotypes using genetic and biochemical techniques, and thereby begin to elucidate the mechanism of vesicle production and regulation. One part of this work elucidated a synthetic genetic growth defect for a strain with reduced OMV production (ΔnlpA, inner membrane lipoprotein with a minor role in methionine transport) and envelope stress (ΔdegP, dual function periplasmic chaperone/ protease responsible for managing proteinaceous waste). This research showed that the growth defect of ΔnlpAΔdegP correlated with reduced OMV production with respect to the hyprevesiculator ΔdegP and the accumulation of protein in the periplasm and DegP substrates in the lumen of OMVs. We further demonstrated that OMVs do not solely act as a stress response pathway to rid the periplasm of otherwise damaging misfolded protein but also of accumulated peptidoglycan (PG) fragments and lipopolysaccharide (LPS), elucidating OMVs as a general stress response pathway critical for bacterial well-being. The second part of this work, focused on the role of PG structure, turnover and covalent crosslinks to the OM in vesiculation. We established a direct link between PG degradation and vesiculation: Mutations in the OM lipoprotein nlpI had been previously established as a very strong hypervesiculation phenotype. In the literature NlpI had been associated with another OM lipoprotein, Spr that was recently identified as a PG hydrolase. The data presented here suggest that NlpI acts as a negative regulator of Spr and that the ΔnlpI hypervesiculation phenotype is a result of rampantly degraded PG by Spr. Additionally, we found that changes in PG structure and turnover correlate with altered vesiculation levels, as well as non-canonical D-amino acids, which are secreted by numerous bacteria on the onset of stationary phase, being a natural factor to increase OMV production. Furthermore, we discovered an inverse relationship between the concentration of Lpp-mediated, covalent crosslinks and the level of OMV production under conditions of modulated PG metabolism and structure. In contrast, situations that lead to periplasmic accumulation (protein, PG fragments, and LPS) and consequent hypervesiculation the overall OM-PG crosslink concentration appears to be unchanged. Form this work, we conclude that multiple pathways lead to OMV production: Lpp concentration-dependent and bulk driven, Lpp concentration-independent.

Division of labor in frontal eye field neurons during presaccadic remapping of visual receptive fields.

Our percept of visual stability across saccadic eye movements may be mediated by presaccadic remapping. Just before a saccade, neurons that remap become visually responsive at a future field (FF), which anticipates the saccade vector. Hence, the neurons use corollary discharge of saccades. Many of the neurons also decrease their response at the receptive field (RF). Presaccadic remapping occurs in several brain areas including the frontal eye field (FEF), which receives corollary discharge of saccades in its layer IV from a collicular-thalamic pathway. We studied, at two levels, the microcircuitry of remapping in the FEF. At the laminar level, we compared remapping between layers IV and V. At the cellular level, we compared remapping between different neuron types of layer IV. In the FEF in four monkeys (Macaca mulatta), we identified 27 layer IV neurons with orthodromic stimulation and 57 layer V neurons with antidromic stimulation from the superior colliculus. With the use of established criteria, we classified the layer IV neurons as putative excitatory (n = 11), putative inhibitory (n = 12), or ambiguous (n = 4). We found that just before a saccade, putative excitatory neurons increased their visual response at the RF, putative inhibitory neurons showed no change, and ambiguous neurons increased their visual response at the FF. None of the neurons showed presaccadic visual changes at both RF and FF. In contrast, neurons in layer V showed full remapping (at both the RF and FF). Our data suggest that elemental signals for remapping are distributed across neuron types in early cortical processing and combined in later stages of cortical microcircuitry.

Dynamics of visual receptive fields in the macaque frontal eye field.

Neuronal receptive fields (RFs) provide the foundation for understanding systems-level sensory processing. In early visual areas, investigators have mapped RFs in detail using stochastic stimuli and sophisticated analytical approaches. Much less is known about RFs in prefrontal cortex. Visual stimuli used for mapping RFs in prefrontal cortex tend to cover a small range of spatial and temporal parameters, making it difficult to understand their role in visual processing. To address these shortcomings, we implemented a generalized linear model to measure the RFs of neurons in the macaque frontal eye field (FEF) in response to sparse, full-field stimuli. Our high-resolution, probabilistic approach tracked the evolution of RFs during passive fixation, and we validated our results against conventional measures. We found that FEF neurons exhibited a surprising level of sensitivity to stimuli presented as briefly as 10 ms or to multiple dots presented simultaneously, suggesting that FEF visual responses are more precise than previously appreciated. FEF RF spatial structures were largely maintained over time and between stimulus conditions. Our results demonstrate that the application of probabilistic RF mapping to FEF and similar association areas is an important tool for clarifying the neuronal mechanisms of cognition.

Similar prevalence and magnitude of auditory-evoked and visually evoked activity in the frontal eye fields: implications for multisensory motor control.

Saccadic eye movements can be elicited by more than one type of sensory stimulus. This implies substantial transformations of signals originating in different sense organs as they reach a common motor output pathway. In this study, we compared the prevalence and magnitude of auditory- and visually evoked activity in a structure implicated in oculomotor processing, the primate frontal eye fields (FEF). We recorded from 324 single neurons while 2 monkeys performed delayed saccades to visual or auditory targets. We found that 64% of FEF neurons were active on presentation of auditory targets and 87% were active during auditory-guided saccades, compared with 75 and 84% for visual targets and saccades. As saccade onset approached, the average level of population activity in the FEF became indistinguishable on visual and auditory trials. FEF activity was better correlated with the movement vector than with the target location for both modalities. In summary, the large proportion of auditory-responsive neurons in the FEF, the similarity between visual and auditory activity levels at the time of the saccade, and the strong correlation between the activity and the saccade vector suggest that auditory signals undergo tailoring to match roughly the strength of visual signals present in the FEF, facilitating accessing of a common motor output pathway.

No self-interaction for two-column massless fields

We investigate the problem of introducing consistent self-couplings in free theories for mixed tensor gauge fields whose symmetry properties are characterized by Young diagrams made of two columns of arbitrary (but different) lengths. We prove that, in flat space, these theories admit no local, Poincaré-invariant, smooth, selfinteracting deformation with at most two derivatives in the Lagrangian. Relaxing the derivative and Lorentz-invariance assumptions, there still is no deformation that modifies the gauge algebra, and in most cases no deformation that alters the gauge transformations. Our approach is based on a Becchi-Rouet-Stora-iyutin (BRST) -cohomology deformation procedure. © 2005 American Institute of Physics.

Parity-violating vertices for spin-3 gauge fields

The problem of constructing consistent parity-violating interactions for spin-3 gauge fields is considered in Minkowski space. Under the assumptions of locality, Poincaré invariance, and parity noninvariance, we classify all the nontrivial perturbative deformations of the Abelian gauge algebra. In space-time dimensions n=3 and n=5, deformations of the free theory are obtained which make the gauge algebra non-Abelian and give rise to nontrivial cubic vertices in the Lagrangian, at first order in the deformation parameter g. At second order in g, consistency conditions are obtained which the five-dimensional vertex obeys, but which rule out the n=3 candidate. Moreover, in the five-dimensional first-order deformation case, the gauge transformations are modified by a new term which involves the second de Wit-Freedman connection in a simple and suggestive way. © 2006 The American Physical Society.

Consistent couplings between fields with a gauge freedom and deformations of the master equation

The antibracket in the antifield-BRST formalism is known to define a map Hp × Hq → Hp + q + 1 associating with two equivalence classes of BRST invariant observables of respective ghost number p and q an equivalence class of BRST invariant observables of ghost number p + q + 1. It is shown that this map is trivial in the space of all functionals, i.e. that its image contains only the zeroth class. However, it is generically non-trivial in the space of local functionals. Implications of this result for the problem of consistent interactions among fields with a gauge freedom are then drawn. It is shown that the obstructions to constructing non-trivial such interactions lie precisely in the image of the antibracket map and are accordingly non-existent if one does not insist on locality. However consistent local interactions are severely constrained. The example of the Chern-Simons theory is considered. It is proved that the only consistent, local, Lorentz covariant interactions for the abelian models are exhausted by the non-abelian Chern-Simons extensions. © 1993.

A Markov modulated Poisson process model for rainfall increments

The problems encountered when using traditional rectangular pulse hierarchical point processmodels for fine temporal resolution and the growing number of available tip-time records suggest that rainfall increments from tipping-bucket gauges be modelled directly. Poisson processes are used with an arrival rate modulated by a Markov chain in Continuous time. The paper shows how, by using two or three states for this chain, much of the structure of the rainfall intensity distribution and the wet/dry sequences can be represented for time-scales as small as 5 minutes.

An explicit scheme for coupling temperature and concentration fields in solidification models

A numerical scheme for coupling temperature and concentration fields in a general solidification model is presented. A key feature of this scheme is an explicit time stepping used in solving the governing thermal and solute conservation equations. This explicit approach results in a local point-by-point coupling scheme for the temperature and concentration and avoids the multi-level iteration required by implicit time stepping schemes. The proposed scheme is validated by predicting the concentration field in a benchmark solidification problem. Results compare well with an available similarity solution. The simplicity of the proposed explicit scheme allows for the incorporation of complex microscale models into a general solidification model. This is demonstrated by investigating the role of dendrite coarsening on the concentration field in the solidification benchmark problem.

Cold crucible melting of reactive metals using combined DC and AC magnetic fields

Cold crucible furnace is widely used for melting reactive metals for high quality castings. Although the water cooled copper crucible avoids contamination, it produces a low superheat of the melt. Experimental and theoretical investigations of the process showed that the increase of the supplied power to the furnace leads to a saturation in the temperature rise of the melt, and no significant increase of the melt superheat can be obtained. The computer model of theprocess has been developed to simulate the time dependent turbulent flow, heat transfer with phase change, and AC and DC magnetohydrodynamics in a time varying liquid metal envelope. The model predicts that the supermimposition of a strong DC field on top of the normal AC field reduces the level of turbulience and stirring in the liquid metal, thereby reducing the heat loss through the base of the crucible and increasing the superheat. The direct measurements of the temperature in the commercial size cold crucbile has confirmed the computer redictions and showed that the addition of a DC field increased the superheat in molten TiAl from ~45C (AC field only) to ~81C (DC+AC fields). The present paper reports further predictions of the effect of a dDC field on top of the AC field and compares these with experimental data.

Droplet generation with modulated AC electromagnetic field at nozzle exit

A method of droplet generation based on applying a modulated AC high frequency magnetic field in the localized region of capillary breakup is considered as ans alternative to traditional methods for high temperature liquid melt droplet serial production by pressure variation. The method is based on a pseudo-spectral approximation with a coordinate transformation adaptin to the developing free surface. The electromagnetic field is recomputed continuously with the domain shape change. Practical application cases for liquid silicon droplets of 0.5 - 2 mm diameter are considered in detail.

The study of flow and temperature fields in conducting droplets suspended in a DC/AC combination field

Electromagnetic Levitation (EML) is a valuable method for measuring the thermo-physical properties of metals - surface tensions, viscosity, thermal/electrical conductivity, specific heat, hemispherical emissivity, etc. – beyond their melting temperature. In EML, a small amount of the test specimen is melted by Joule heating in a suspended AC coil. Once in liquid state, a small perturbation causes the liquid envelope to oscillate and the frequency of oscillation is then used to compute its surface tension by the well know Rayleigh formula. Similarly, the rate at which the oscillation is dampened relates to the viscosity. To measure thermal conductivity, a sinusoidally varying laser source may be used to heat the polar axis of the droplet and the temperature response measured at the polar opposite – the resulting phase shift yields thermal conductivity. All these theoretical methods assume that convective effects due to flow within the droplet are negligible compared to conduction, and similarly that the flow conditions are laminar; a situation that can only be realised under microgravity conditions. Hence the EML experiment is the method favoured for Spacelab experiments (viz. TEMPUS). Under terrestrial conditions, the full gravity force has to be countered by a much larger induced magnetic field. The magnetic field generates strong flow within the droplet, which for droplets of practical size becomes irrotational and turbulent. At the same time the droplet oscillation envelope is no longer ellipsoidal. Both these conditions invalidate simple theoretical models and prevent widespread EML use in terrestrial laboratories. The authors have shown in earlier publications that it is possible to suppress most of the turbulent convection generated in the droplet skin layer, through use of a static magnetic field. Using a pseudo-spectral discretisation method it is possible compute very accurately the dynamic variation in the suspended fluid envelope and simultaneously compute the time-varying electromagnetic, flow and thermal fields. The use of a DC field as a dampening agent was also demonstrated in cold crucible melting, where suppression of turbulence was achieved in a much larger liquid metal volume and led to increased superheat in the melt and reduction of heat losses to the water-cooled walls. In this paper, the authors describe the pseudo-spectral technique as applied to EML to compute the combined effects of AC and DC fields, accounting for all the flow-induced forces acting on the liquid volume (Lorentz, Maragoni, surface tension, gravity) and show example simulations.