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A primary neural cell culture model to study neuron, astrocyte, and microglia interactions in neuroinflammation.
BackgroundInteractions between neurons, astrocytes, and microglia critically influence neuroinflammatory responses to insult in the central nervous system. In vitro astrocyte and microglia cultures are powerful tools to study specific molecular pathways involved in neuroinflammation; however, in order to better understand the influence of cellular crosstalk on neuroinflammation, new multicellular culture models are required.MethodsPrimary cortical cells taken from neonatal rats were cultured in a serum-free "tri-culture" medium formulated to support neurons, astrocytes, and microglia, or a "co-culture" medium formulated to support only neurons and astrocytes. Caspase 3/7 activity and morphological changes were used to quantify the response of the two culture types to different neuroinflammatory stimuli mimicking sterile bacterial infection (lipopolysaccharide (LPS) exposure), mechanical injury (scratch), and seizure activity (glutamate-induced excitotoxicity). The secreted cytokine profile of control and LPS-exposed co- and tri-cultures were also compared.ResultsThe tri-culture maintained a physiologically relevant representation of neurons, astrocytes, and microglia for 14âdays in vitro, while the co-cultures maintained a similar population of neurons and astrocytes, but lacked microglia. The continuous presence of microglia did not negatively impact the overall health of the neurons in the tri-culture, which showed reduced caspase 3/7 activity and similar neurite outgrowth as the co-cultures, along with an increase in the microglia-secreted neurotrophic factor IGF-1 and a significantly reduced concentration of CX3CL1 in the conditioned media. LPS-exposed tri-cultures showed significant astrocyte hypertrophy, increase in caspase 3/7 activity, and the secretion of a number of pro-inflammatory cytokines (e.g., TNF, IL-1α, IL-1ÎČ, and IL-6), none of which were observed in LPS-exposed co-cultures. Following mechanical trauma, the tri-culture showed increased caspase 3/7 activity, as compared to the co-culture, along with increased astrocyte migration towards the source of injury. Finally, the microglia in the tri-culture played a significant neuroprotective role during glutamate-induced excitotoxicity, with significantly reduced neuron loss and astrocyte hypertrophy in the tri-culture.ConclusionsThe tri-culture consisting of neurons, astrocytes, and microglia more faithfully mimics in vivo neuroinflammatory responses than standard mono- and co-cultures. This tri-culture can be a useful tool to study neuroinflammation in vitro with improved accuracy in predicting in vivo neuroinflammatory phenomena
On the recombination in high-order harmonic generation in molecules
We show that the dependence of high-order harmonic generation (HHG) on the
molecular orientation can be understood within a theoretical treatment that
does not involve the strong field of the laser. The results for H_2 show
excellent agreement with time-dependent strong field calculations for model
molecules, and this motivates a prediction for the orientation dependence of
HHG from the N_2 3s_g valence orbital. For both molecules, we find that the
polarization of recombination photons is influenced by the molecular
orientation. The variations are particularly pronounced for the N_2 valence
orbital, which can be explained by the presence of atomic p-orbitals.Comment: 6 pages 7 figure
Semiclassical two-step model for strong-field ionization
We present a semiclassical two-step model for strong-field ionization that
accounts for path interferences of tunnel-ionized electrons in the ionic
potential beyond perturbation theory. Within the framework of a classical
trajectory Monte-Carlo representation of the phase-space dynamics, the model
employs the semiclassical approximation to the phase of the full quantum
propagator in the exit channel. By comparison with the exact numerical solution
of the time-dependent Schr\"odinger equation for strong-field ionization of
hydrogen, we show that for suitable choices of the momentum distribution after
the first tunneling step, the model yields good quantitative agreement with the
full quantum simulation. The two-dimensional photoelectron momentum
distributions, the energy spectra, and the angular distributions are found to
be in good agreement with the corresponding quantum results. Specifically, the
model quantitatively reproduces the fan-like interference patterns in the
low-energy part of the two-dimensional momentum distributions as well as the
modulations in the photoelectron angular distributions.Comment: 31 pages, 7 figure
Initial-state dependence in time-dependent density functional theory
Time-dependent density functionals in principle depend on the initial state
of the system, but this is ignored in functional approximations presently in
use. For one electron it is shown there is no initial-state dependence: for any
density, only one initial state produces a well-behaved potential. For two
non-interacting electrons with the same spin in one-dimension, an initial
potential that makes an alternative initial wavefunction evolve with the same
density and current as a ground state is calculated. This potential is
well-behaved and can be made arbitrarily different from the original potential
Non-sequential triple ionization in strong fields
We consider the final stage of triple ionization of atoms in a strong
linearly polarized laser field. We propose that for intensities below the
saturation value for triple ionization the process is dominated by the
simultaneous escape of three electrons from a highly excited intermediate
complex. We identify within a classical model two pathways to triple
ionization, one with a triangular configuration of electrons and one with a
more linear one. Both are saddles in phase space. A stability analysis
indicates that the triangular configuration has the larger cross sections and
should be the dominant one. Trajectory simulations within the dominant symmetry
subspace reproduce the experimentally observed distribution of ion momenta
parallel to the polarization axis.Comment: 9 pages, 8 figures, accepted for publication in Phys. Rev.
DeepBrain: Functional Representation of Neural In-Situ Hybridization Images for Gene Ontology Classification Using Deep Convolutional Autoencoders
This paper presents a novel deep learning-based method for learning a
functional representation of mammalian neural images. The method uses a deep
convolutional denoising autoencoder (CDAE) for generating an invariant, compact
representation of in situ hybridization (ISH) images. While most existing
methods for bio-imaging analysis were not developed to handle images with
highly complex anatomical structures, the results presented in this paper show
that functional representation extracted by CDAE can help learn features of
functional gene ontology categories for their classification in a highly
accurate manner. Using this CDAE representation, our method outperforms the
previous state-of-the-art classification rate, by improving the average AUC
from 0.92 to 0.98, i.e., achieving 75% reduction in error. The method operates
on input images that were downsampled significantly with respect to the
original ones to make it computationally feasible
Developmental Exposure to Polychlorinated Biphenyls Influences Stroke Outcome in Adult Rats
BackgroundThe "developmental origins of adult disease" hypothesis was originally derived from evidence linking low birth weight to cardiovascular diseases including stroke. Subsequently, it has been expanded to include developmental exposures to environmental contaminants as risk factors for adult onset disease.ObjectiveOur goal in this study was to test the hypothesis that developmental exposure to poly-chlorinated biphenyls (PCBs) alters stroke outcome in adults.MethodsWe exposed rats to the PCB mixture Aroclor 1254 (A1254) at 0.1 or 1 mg/kg/day in the maternal diet throughout gestation and lactation. Focal cerebral ischemia was induced at 6-8 weeks of age via middle cerebral artery occlusion, and infarct size was measured in the cerebral cortex and striatum at 22 hr of reperfusion. PCB congeners were quantified in brain tissue by gas chromatography with microelectron capture detection, and cortical and striatal expression of Bcl2 and Cyp2C11 were quantified by quantitative reverse transcriptase-polymerase chain reaction.ResultsDevelopmental exposure to A1254 significantly decreased striatal infarct in females and males at 0.1 and 1 mg/kg/day, respectively. Predominantly ortho-substituted PCB congeners were detected above background levels in brains of adult females and males exposed to A1254 at 1 but not 0.1 mg/kg/day. Effects of developmental A1254 exposure on Bcl2 and Cyp2C11 expression did not correlate with effects on infarct volume.ConclusionOur data provide proof of principle that developmental exposures to environmental contaminants influence the response of the adult brain to ischemic injury and thus represent potentially important determinants of stroke susceptibility
Correlated electron emission in laser-induced nonsequence double ionization of Helium
In this paper, we have investigated the correlated electron emission of the
nonsequence double ionization (NSDI) in an intense linearly polarized field.
The theoretical model we employed is the semiclassical rescattering model, the
model atom we used is the helium. We find a significant correlation between
magnitude and direction of the momentum of two emission electrons, and give a
good explanation for this striking phenomenon by observing the classical
collisional trajectories. We argue that this correlation phenomenon is
universal in NSDI process, as revealed by the recent experiment on the argon.Comment: 4 pages, 3 figures, accepted for publication in Phys. Rev.
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