Delineating neuroinflammation, parasite CNS invasion, and blood-brain barrier dysfunction in an experimental murine model of human African trypanosomiasis

Although Trypanosoma brucei spp. was first detected by Aldo Castellani in CSF samples taken from sleeping sickness patients over a century ago there is still a great deal of debate surrounding the timing, route and effects of transmigration of the parasite from the blood to the CNS. In this investigation, we have applied contrast-enhance magnetic resonance imaging (MRI) to study the effects of trypanosome infection on the blood-brain barrier (BBB) in the well-established GVR35 mouse model of sleeping sickness. In addition, we have measured the trypanosome load present in the brain using quantitative Taqman PCR and assessed the severity of the neuroinflammatory reaction at specific time points over the course of the infection. Contrast enhanced – MRI detected a significant degree of BBB impairment in mice at 14 days following trypanosome infection, which increased in a step-wise fashion as the disease progressed. Parasite DNA was present in the brain tissue on day 7 after infection. This increased significantly in quantity by day 14 post-infection and continued to rise as the infection advanced. A progressive increase in neuroinflammation was detected following trypanosome infection, reaching a significant level of severity on day 14 post-infection and rising further at later time-points. In this model stage-2 disease presents at 21 days post-infection. The combination of the three methodologies indicates that changes in the CNS become apparent prior to the onset of established stage-2 disease. This could in part account for the difficulties associated with defining specific criteria to distinguish stage-1 and stage-2 infections and highlights the need for improved staging diagnostics

Trypanosoma brucei brucei invasion and T-cell infiltration of the brain parenchyma in experimental sleeping sickness: timing and correlation with functional changes

Background: The timing of Trypanosoma brucei entry into the brain parenchyma to initiate the second, meningoencephalitic stage of human African trypanosomiasis or sleeping sickness is currently debated and even parasite invasion of the neuropil has been recently questioned. Furthermore, the relationship between neurological features and disease stage are unclear, despite the important diagnostic and therapeutic implications. Methodology: Using a rat model of chronic Trypanosoma brucei brucei infection we determined the timing of parasite and T-cell neuropil infiltration and its correlation with functional changes. Parasite DNA was detected using trypanosome-specific PCR. Body weight and sleep structure alterations represented by sleep-onset rapid eye movement (SOREM) periods, reported in human and experimental African trypanosomiasis, were monitored. The presence of parasites, as well as CD4+ and CD8+ T-cells in the neuropil was assessed over time in the brain of the same animals by immunocytochemistry and quantitative analyses. Principal findings: Trypanosome DNA was present in the brain at day 6 post-infection and increased more than 15-fold by day 21. Parasites and T-cells were observed in the parenchyma from day 9 onwards. Parasites traversing blood vessel walls were observed in the hypothalamus and other brain regions. Body weight gain was reduced from day 7 onwards. SOREM episodes started in most cases early after infection, with an increase in number and duration after parasite neuroinvasion. Conclusion: These findings demonstrate invasion of the neuropil over time, after an initial interval, by parasites and lymphocytes crossing the blood-brain barrier, and show that neurological features can precede this event. The data thus challenge the current clinical and cerebrospinal fluid criteria of disease staging

Microarray profiling predicts early neurological and immune phenotypic traits in advance of CNS disease during disease progression in Trypanosoma.b.brucei infected CD1 mouse brains

Human African trypanosomiasis (HAT), also known as sleeping sickness, is a major cause of mortality and morbidity in sub-Saharan Africa. We hypothesised that recent findings of neurological features and parasite brain infiltration occurring at much earlier stages in HAT than previously thought could be explained by early activation of host genetic programmes controlling CNS disease. Accordingly, a transcriptomal analysis was performed on brain tissue at 0, 7, 14, 21 and 28dpi from the HAT CD1/GVR35 mouse model. Up to 21dpi, most parasites are restricted to the blood and lymphatic system. Thereafter the trypanosomes enter the brain initiating the encephalitic stage. Analysis of ten different time point Comparison pairings, revealed a dynamic transcriptome comprising four message populations. All 7dpi Comparisons had by far more differentially expressed genes compared to all others. Prior to invasion of the parenchyma, by 7dpi, ~2,000 genes were up-regulated, denoted [7dpi↑] in contrast to a down regulated population [7dpi↓] also numbering ~2,000. However, by 14dpi both patterns had returned to around the pre-infected levels. The third, [28dpi↑] featured over three hundred transcripts which had increased modestly up to14dpi, thereafter were significantly up-regulated and peaked at 28dpi. The fourth, a minor population, [7dpi↑-28dpi↑], had similar elevated levels at 7dpi and 28dpi. KEGG and GO enrichment analysis predicted a diverse phenotype by 7dpi with changes to innate and adaptive immunity, a Type I interferon response, neurotransmission, synaptic plasticity, pleiotropic signalling, circadian activity and vascular permeability without disruption of the blood brain barrier. This key observation is consistent with recent rodent model neuroinvasion studies and clinical reports of Stage 1 HAT patients exhibiting CNS symptoms. Together, these findings challenge the strict Stage1/Stage2 phenotypic demarcation in HAT and show that that significant neurological, and immune changes can be detected prior to the onset of CNS disease

Cosmology, Particle Physics and Superfluid 3He

Many direct parallels connect superfluid 3He with the field theories describing the physical vacuum, gauge fields and elementary fermions. Superfluid $^3$He exhibits a variety of topological defects which can be detected with single-defect sensitivity. Modern scenarios of defect-mediated baryogenesis can be simulated by the interaction of the 3He vortices and domain walls with fermionic quasiparticles. Formation of defects in a symmetry-breaking phase transition in the early Universe, which could be responsible for large-scale structure formation and for microwave-background anisotropy, also may be modelled in the laboratory. This is supported by the recent observation of vortex formation in neutron-irradiated 3He-B where the "primordial fireball" is formed in an exothermic nuclear reaction.Comment: Invited talk at LT-21 Conference, 20 pages, 3 figures available at request, compressed ps file of the camera-ready format with 3 figures is at ftp://boojum.hut.fi/pub/publications/lowtemp/LTL-96006.ps.g

Natural Orbitals and BEC in traps, a diffusion Monte Carlo analysis

We investigate the properties of hard core Bosons in harmonic traps over a wide range of densities. Bose-Einstein condensation is formulated using the one-body Density Matrix (OBDM) which is equally valid at low and high densities. The OBDM is calculated using diffusion Monte Carlo methods and it is diagonalized to obtain the "natural" single particle orbitals and their occupation, including the condensate fraction. At low Boson density, $na^3 < 10^{-5}$, where $n = N/V$ and $a$ is the hard core diameter, the condensate is localized at the center of the trap. As $na^3$ increases, the condensate moves to the edges of the trap. At high density it is localized at the edges of the trap. At $na^3 \leq 10^{-4}$ the Gross-Pitaevskii theory of the condensate describes the whole system within 1%. At $na^3 \approx 10^{-3}$ corrections are 3% to the GP energy but 30% to the Bogoliubov prediction of the condensate depletion. At $na^3 \gtrsim 10^{-2}$, mean field theory fails. At $na^3 \gtrsim 0.1$, the Bosons behave more like a liquid $^4$He droplet than a trapped Boson gas.Comment: 13 pages, 14 figures, submitted Phys. Rev.

Transport spin polarization of Ni_xFe_{1-x}: electronic kinematics and band structure

We present measurements of the transport spin polarization of Ni_xFe_{1-x} (0<x<1) using the recently-developed Point Contact Andreev Reflection technique, and compare them with our first principles calculations of the spin polarization for this system. Surpisingly, the measured spin polarization is almost composition-independent. The results clearly demonstrate that the sign of the transport spin polarization does not coincide with that of the difference of the densities of states at the Fermi level. Calculations indicate that the independence of the spin polarization of the composition is due to compensation of density of states and Fermi velocity in the s- and d- bands

Thermal Detection of Turbulent and Laminar Dissipation in Vortex Front Motion

We report on direct measurements of the energy dissipated in the spin-up of the superfluid component of 3He-B. A vortex-free sample is prepared in a cylindrical container, where the normal component rotates at constant angular velocity. At a temperature of 0.20Tc, seed vortices are injected into the system using the shear-flow instability at the interface between 3He-B and 3He-A. These vortices interact and create a turbulent burst, which sets a propagating vortex front into motion. In the following process, the free energy stored in the initial vortex-free state is dissipated leading to the emission of thermal excitations, which we observe with a bolometric measurement. We find that the turbulent front contains less than the equilibrium number of vortices and that the superfluid behind the front is partially decoupled from the reference frame of the container. The final equilibrium state is approached in the form of a slow laminar spin-up as demonstrated by the slowly decaying tail of the thermal signal.Comment: 12 pages, 5 figures, to appear in Journal of Low Temperature Physic

Coherent matter wave inertial sensors for precision measurements in space

We analyze the advantages of using ultra-cold coherent sources of atoms for matter-wave interferometry in space. We present a proof-of-principle experiment that is based on an analysis of the results previously published in [Richard et al., Phys. Rev. Lett., 91, 010405 (2003)] from which we extract the ratio h/m for 87Rb. This measurement shows that a limitation in accuracy arises due to atomic interactions within the Bose-Einstein condensate

Quantum saturation and condensation of excitons in Cu2_2O: a theoretical study

Recent experiments on high density excitons in Cu$_2$O provide evidence for degenerate quantum statistics and Bose-Einstein condensation of this nearly ideal gas. We model the time dependence of this bosonic system including exciton decay mechanisms, energy exchange with phonons, and interconversion between ortho (triplet-state) and para (singlet-state) excitons, using parameters for the excitonic decay, the coupling to acoustic and low-lying optical phonons, Auger recombination, and ortho-para interconversion derived from experiment. The single adjustable parameter in our model is the optical-phonon cooling rate for Auger and laser-produced hot excitons. We show that the orthoexcitons move along the phase boundary without crossing it (i.e., exhibit a ``quantum saturation''), as a consequence of the balance of entropy changes due to cooling of excitons by phonons and heating by the non-radiative Auger two-exciton recombination process. The Auger annihilation rate for para-para collisions is much smaller than that for ortho-para and ortho-ortho collisions, explaining why, under the given experimental conditions, the paraexcitons condense while the orthoexcitons fail to do so.Comment: Revised to improve clarity and physical content 18 pages, revtex, figures available from G. Kavoulakis, Physics Department, University of Illinois, Urban

Explosion Flame Acceleration over Obstacles: Effects of Separation Distance for a Range of Scales

The influence of obstacle separation distance on explosion flame acceleration was studied for 10% methane-air mixtures using two 20% blockage obstacles with variable number and width of bars (variable obstacle length scale) were investigated in a 162 mm diameter 4.5 m long tube with ignition on the centre of the closed end and flame propagation towards the open end. The spacing between the obstacles was varied from 0.25 m to 2.75 m. It was observed that the maximum overpressure and flame speed increased with the reduction in number of flat-bars (i.e. with increasing obstacle length scale). A maximum overpressure of 129 kPa at 2.25 m obstacle spacing was achieved with 1-flat-bar obstacles, followed by 118 kPa and 110 kPa for 2 and 4-flat-bars respectively at 1.25 m and 0.5 m obstacle separation. Turbulent to laminar burning velocity ratios downstream of the second obstacle at the optimum spacing for maximum interaction were in the range of 62-122. These are the magnitudes of flame acceleration required to explain overpressures in vapour cloud explosions in the presence of obstacles. It is worth appreciating that two obstacles of lower blockages but spaced optimally could generate higher explosion severity in terms of overpressure, flame speed and turbulence level similar to real gas explosion incidents