Stimulus Discrimination in Cerebellar Purkinje Neurons

Cerebellar Purkinje neurons fire spontaneously in the absence of synaptic input. Overlaid on this intrinsic activity, excitatory input from parallel fibres can add simple spikes to the output train, whereas inhibitory input from interneurons can introduce pauses. These and other influences lead to an irregular spike train output in Purkinje neurons in vitro and in vivo, supplying a variable inhibitory drive to deep cerebellar nuclear neurons. From a computational perspective, this variability raises some questions, as individual spikes induced by excitatory inputs are indistinguishable from intrinsic firing activity. Although bursts of high-frequency excitatory input could be discriminated unambiguously from background activity, granule neurons are known to fire in vivo over a wide range of frequencies. This would mean that much of the sensory information relayed through the cerebellar cortex would be lost within the random variation in background activity. We speculated that alternative mechanisms for signal discrimination may exist, and sought to identify characteristic motifs within the sequence of spikes that followed stimulation events. We found that under certain conditions, parallel fibre stimulation could reliably add a “couplet” of spikes with an unusually short interspike interval to the output train. Therefore, despite representing a small fraction of the total number of spikes, these signals can be reliably discriminated from background firing on a moment-to-moment basis, and could result in a differential downstream response

North Atlantic climate variability from a self-organizing map perspective

[1] North Atlantic variability in general, and the North Atlantic Oscillation (NAO) in particular, is a long-studied, very important but still not well-understood problem in climatology. The recent trend to a higher wintertime NAO index was accompanied by an additional increase in the Azores High not coupled to changes in the Icelandic Low, as shown by a self-organizing maps (SOMs) analysis of monthly mean DJF mean sea level pressure data from 1957 to 2002. SOMs are a nonlinear tool to optimally extract a user-specified number of patterns or icons from an input data set and to uniquely relate any input data field to an icon, allowing analyses of occurrence frequencies and transitions complementary to principal component analysis (PCA). SOMs analysis of ERA-40 data finds a North Atlantic monopole roughly colocated with the mean position of the Azores High, as well as the well-known NAO dipole involving the Icelandic Low and the subtropical high. Little trend is shown in December, but the Azores High increased along with the NAO in January and February over the study interval, with implications for storminess in northwestern Europe. In short, our SOM-based analyses of winter MSLP have both confirmed prior knowledge and expanded it through the relative ease of use and power with nonlinear systems of the SOM-based approach to climatological analysis

Evaluation of an Electrolyte Analyser for Measurement of Concentrations of Ionized Calcium and Magnesium in Cats

The goal of this study was to evaluate the Nova CRT 8 electrolyte analyser for determination of concentrations of ionized calcium (Cai) and magnesium (Mgi) in cats, todetermine the effects of sample handling and storage and to establish reference ranges. The precision and analytical accuracy of the Nova CRT 8 analyser were good. The concentrations of Cai and Mgi were significantly lower in aerobically handled serum samples than in those handled anaerobically. The concentrations of Cai and Mgi differed significantly among whole blood, plasma and serum. In anaerobically handled serum, the concentration of Cai was stable for 8 h at 22°C, for 5 days at 4°C and for 1 week at −20°C. The concentration of Mgi was stable for 4 h at 22°C but for less than 24 h at 4°C and for less than 1 week at −20°C. In serum from 36 cats, the reference ranges were 1.20-1.35 mmol/L for Cai and 0.47-0.59 mmol/L for Mgi. The Nova CRT 8 electrolyte analyser is suitable for determination of Cai and Mgi concentrations in cats. Anaerobically handled serum samples are recommended and, stored at room temperature, they yield accurate results when analysed within 4

Unified description of floppy and rigid rotating Wigner molecules formed in quantum dots

Restoration of broken circular symmetry is used to explore the characteristics of the ground states and the excitation spectra of rotating Wigner molecules (RWM's) formed in two-dimensional parabolic N-electron quantum dots. In high magnetic fields, the RWM's are floppy rotors with the energies of the magic angular momentum (L) states obeying aL + b/L^{1/2}. Under such fields the ground-state energies (referenced to the kinetic energy in the lowest Landau level) approach the electrostatic energy of N point charges in the classical equilibrium molecular configuration. At zero field and strong interelectron repulsion, the RWM's behave like quasiclassical rigid rotors whose energies vary as L^2. The particular L-dependence in high B is inherent and natural to a floppy rotating WM, and it can be used as a crucial diagnostic tool for resolving the recently posed question whether the composite-fermion or the RWM picture is appropriate for QD's.Comment: 5 pages. Revtex4 with 3 EPS figures and 2 tables . For related papers, see http://www.prism.gatech.edu/~ph274c

Neutralizing monoclonal antibodies define two different functional sites in human interleukin-4

Human interleukin-4 (IL-4) is a small four-helix-bundle protein which is essential for organizing defense reactions against macroparasites, in particular helminths. Human IL-4 also appears to exert a pathophysiological role during various IgE-mediated allergic diseases. Seven different monoclonal antibodies neutralizing the activity of human IL-4 were studied in order to identify functionally important epitopes. A collection of 41 purified IL-4 variants was used to analyse how defined amino acid replacements affect binding affinity for each individual mAb. Specific amino acid positions could be assigned to four different epitopes. mAbs recognizing epitopes on helix A and/or C interfered with IL-4 receptor binding and thus inhibited IL-4 function. However, other mAbs also inhibiting IL-4 function recognized an epitope on helix D of IL-4 and did not inhibit IL-4 binding to the receptor protein. One mAb, recognizing N-terminal and C-terminal residues, partially competed for binding to the receptor. The results of these mAb epitope analyses confirm and extend previous data on the functional consequences of the amino acid replacements which showed that amino acid residues in helices A and C of IL-4 provide a binding site for the cloned IL-4 receptor and that a signalling site in helix D interacts with a further receptor protein

Wind energy production in forests conflicts with tree-roosting bats

Many countries are investing heavily in wind power generation,1 triggering a high demand for suitable land. As a result, wind energy facilities are increasingly being installed in forests,2,3 despite the fact that forests are crucial for the protection of terrestrial biodiversity.4 This green-green dilemma is particularly evident for bats, as most species at risk of colliding with wind turbines roost in trees.2 With some of these species reported to be declining,5,6,7,8 we see an urgent need to understand how bats respond to wind turbines in forested areas, especially in Europe where all bat species are legally protected. We used miniaturized global positioning system (GPS) units to study how European common noctule bats (Nyctalus noctula), a species that is highly vulnerable at turbines,9 respond to wind turbines in forests. Data from 60 tagged common noctules yielded a total of 8,129 positions, of which 2.3% were recorded at distances <100 m from the nearest turbine. Bats were particularly active at turbines <500 m near roosts, which may require such turbines to be shut down more frequently at times of high bat activity to reduce collision risk. Beyond roosts, bats avoided turbines over several kilometers, supporting earlier findings on habitat loss for forest-associated bats.10 This habitat loss should be compensated by developing parts of the forest as refugia for bats. Our study highlights that it can be particularly challenging to generate wind energy in forested areas in an ecologically sustainable manner with minimal impact on forests and the wildlife that inhabit them

Probabilistic encoding of stimulus strength in astrocyte global calcium signals

Astrocyte calcium signals can range in size from subcellular microdomains to waves that spread through the whole cell (and into connected cells). The differential roles of such local or global calcium signaling are under intense investigation, but the mechanisms by which local signals evolve into global signals in astrocytes are not well understood, nor are the computational rules by which physiological stimuli are transduced into a global signal. To investigate these questions, we transiently applied receptor agonists linked to calcium signaling to primary cultures of cerebellar astrocytes. Astrocytes repetitively tested with the same stimulus responded with global signals intermittently, indicating that each stimulus had a defined probability for triggering a response. The response probability varied between agonists, increased with agonist concentration, and could be positively and negatively modulated by crosstalk with other signaling pathways. To better understand the processes determining the evolution of a global signal, we recorded subcellular calcium “puffs” throughout the whole cell during stimulation. The key requirement for puffs to trigger a global calcium wave following receptor activation appeared to be the synchronous release of calcium from three or more sites, rather than an increasing calcium load accumulating in the cytosol due to increased puff size, amplitude, or frequency. These results suggest that the concentration of transient stimuli will be encoded into a probability of generating a global calcium response, determined by the likelihood of synchronous release from multiple subcellular sites

Evaluation of cortisol precursors for the diagnosis of pituitary-dependent hypercortisolism in dogs

The serum concentrations of cortisol, 17alpha-hydroxypregnenolone, 17alpha-hydroxyprogesterone, 21-deoxycortisol and 11-deoxycortisol were measured in 19 healthy dogs, 15 dogs with pituitary-dependent hypercortisolism (pdh) and eight dogs with other diseases before and one hour after an injection of synthetic adrenocorticotrophic hormone (acth). At both times the dogs with pdh had significantly higher concentrations of cortisol, 17alpha-hydroxypregnenolone, 17alpha-hydroxyprogesterone and 21-deoxycortisol than the healthy dogs. Basal 11-deoxycortisol concentrations were also significantly higher in dogs with pdh compared with healthy dogs. When compared with the dogs with other diseases, the dogs with pdh had significantly higher basal and post-acth cortisol and basal 21-deoxycortisol, and significantly lower post-acth 11-deoxycortisol concentrations. The dogs with other diseases had significantly higher post-acth cortisol, 17alpha-hydroxyprogesterone and 11-deoxycortisol concentrations than the healthy dogs. In general, the post-acth concentrations of 17alpha-hydroxypregnenolone, 17alpha-hydroxyprogesterone, 11-deoxycortisol and 21-deoxycortisol were more variable than the post-acth concentrations of cortisol, resulting in large overlaps of the concentrations of these hormones between the three groups. A two-graph receiver operating characteristic (ROC) analysis was used to maximise the sensitivity and specificity of each hormone for diagnosing hypercortisolism; it showed that the post-acth concentration of cortisol had the highest sensitivity and specificity. The overlaps between the healthy dogs, the dogs with pdh and the dogs with other diseases suggested that the individual precursor hormones would not be useful as a screening test for hypercortisolism

Ca2+-controlled competitive diacylglycerol binding of protein kinase C isoenzymes in living cells

The cellular decoding of receptor-induced signaling is based in part on the spatiotemporal activation pattern of PKC isoforms. Because classical and novel PKC isoforms contain diacylglycerol (DAG)-binding C1 domains, they may compete for DAG binding. We reasoned that a Ca2+-induced membrane association of classical PKCs may accelerate the DAG binding and thereby prevent translocation of novel PKCs. Simultaneous imaging of fluorescent PKC fusion proteins revealed that during receptor stimulation, PKCα accumulated in the plasma membrane with a diffusion-limited kinetic, whereas translocation of PKCɛ was delayed and attenuated. In BAPTA-loaded cells, however, a selective translocation of PKCɛ, but not of coexpressed PKCα, was evident. A membrane-permeable DAG analogue displayed a higher binding affinity for PKCɛ than for PKCα. Subsequent photolysis of caged Ca2+ immediately recruited PKCα to the membrane, and DAG-bound PKCɛ was displaced. At low expression levels of PKCɛ, PKCα concentration dependently prevented the PKCɛ translocation with half-maximal effects at equimolar coexpression. Furthermore, translocation of endogenous PKCs in vascular smooth muscle cells corroborated the model that a competition between PKC isoforms for DAG binding occurs at native expression levels. We conclude that Ca2+-controlled competitive DAG binding contributes to the selective recruitment of PKC isoforms after receptor activation