Vacuolar ATPase depletion contributes to dysregulation of endocytosis in bloodstream forms of Trypanosoma brucei

BACKGROUND Vacuolar H-ATPase (V-ATPase) is a highly conserved protein complex which hydrolyzes ATP and pumps protons to acidify vacuolar vesicles. Beyond its role in pH maintenance, the involvement of V-ATPase in endocytosis is well documented in mammals and plants but is less clear in Trypanosoma brucei. METHODS In this study, the subcellular localization of V-ATPase subunit B (TbVAB) of T. brucei was assessed via in situ N-terminal YFP-tagging and immunofluorescence assays. Transgenic bloodstream forms (BSF) of T. brucei were generated which comprised either a V-ATPase subunit B (TbVAB) conditional knockout or a V-ATPase subunit A (TbVAA) knockdown. Acridine orange and BCECF-AM were employed to assess the roles of V-ATPase in the pH regulation of BSF T. brucei. The endocytic activities of three markers were also characterized by flow cytometry analyses. Furthermore, trypanosomes were counted from trypanolysis treatment groups (either containing 1% or 5% NHS) and endocytosed trypanosome lytic factor (TLF) was also analyzed by an immunoblotting assay. RESULTS TbVAB was found to localize to acidocalcisomes, lysosomes and probably also to endosomes of BSF of T. brucei and was demonstrated to be essential for cell growth. TbVAB depletion neutralized acidic organelles at 24 hours post-tetracycline depletion (hpd), meanwhile the steady state intracellular pH increased from 7.016 ± 0.013 to 7.422 ± 0.058. Trypanosomes with TbVAB depletion at 24 hpd were found to take up more transferrin (2.068 ± 0.277 fold) but less tomato lectin (49.31 ± 22.57%) by endocytosis, while no significant change was detected in dextran uptake. Similar endocytic dysregulated phenotypes were also observed in TbVAA knockdown cells. In addition, TbVAB depleted trypanosomes showed a low uptake of TLF and exhibited less sensitive to lysis in both 1% and 5% NHS treatments. CONCLUSIONS TbVAB is a key component of V-ATPase and was found to play a key function in endocytosis as well as exhibiting different effects in a receptor/cargo dependent manner in BSF of T. brucei. Besides vacuolar alkalinization, the dysregulation of endocytosis in TbVAB depleted T. brucei is considered to contribute to the reduced sensitivity to lysis by normal human serum

Dynamics of inelastically colliding rough spheres: Relaxation of translational and rotational energy

We study the exchange of kinetic energy between translational and rotational degrees of freedom for inelastic collisions of rough spheres. Even if equipartition holds in the initial state it is immediately destroyed by collisions. The simplest generalisation of the homogeneous cooling state allows for two temperatures, characterizing translational and rotational degrees of freedom separately. For times larger than a crossover frequency, which is determined by the Enskog frequency and the initial temperature, both energies decay algebraically like $t^{-2}$ with a fixed ratio of amplitudes, different from one.Comment: 5 pages, RevTeX, 2 eps figures, slightly expanded discussion, new figures with dimensionless units, added references, accepted for publication in PRE as a Rapid Com

First and Second Sound Modes of a Bose-Einstein Condensate in a Harmonic Trap

We have calculated the first and second sound modes of a dilute interacting Bose gas in a spherical trap for temperatures ($0.6<T/T_{c}<1.2$) and for systems with $10^4$ to $10^8$ particles. The second sound modes (which exist only below $T_{c}$) generally have a stronger temperature dependence than the first sound modes. The puzzling temperature variations of the sound modes near $T_{c}$ recently observed at JILA in systems with $10^3$ particles match surprisingly well with those of the first and second sound modes of much larger systems.Comment: a shorten version, more discussions are given on the nature of the second sound. A long footnote on the recent work of Zaremba, Griffin, and Nikuni (cond-mat/9705134) is added, the spectrum of the (\ell=1, n_2=0) mode is included in fig.

Homogeneous cooling of rough, dissipative particles: Theory and simulations

We investigate freely cooling systems of rough spheres in two and three dimensions. Simulations using an event driven algorithm are compared with results of an approximate kinetic theory, based on the assumption of a generalized homogeneous cooling state. For short times $t$, translational and rotational energy are found to change linearly with $t$. For large times both energies decay like $t^{-2}$ with a ratio independent of time, but not corresponding to equipartition. Good agreement is found between theory and simulations, as long as no clustering instability is observed. System parameters, i.e. density, particle size, and particle mass can be absorbed in a rescaled time, so that the decay of translational and rotational energy is solely determined by normal restitution and surface roughness.Comment: 10 pages, 10 eps-figure

Spinor Bose Condensates in Optical Traps

In an optical trap, the ground state of spin-1 Bosons such as $^{23}$Na, $^{39}$K, and $^{87}$Rb can be either a ferromagnetic or a "polar" state, depending on the scattering lengths in different angular momentum channel. The collective modes of these states have very different spin character and spatial distributions. While ordinary vortices are stable in the polar state, only those with unit circulation are stable in the ferromagnetic state. The ferromagnetic state also has coreless (or Skyrmion) vortices like those of superfluid $^{3}$He-A. Current estimates of scattering lengths suggest that the ground states of $^{23}$Na and $^{87}$Rb condensate are a polar state and a ferromagnetic state respectively.Comment: 11 pages, no figures. email : [email protected]

Granular cooling of hard needles

We have developed a kinetic theory of hard needles undergoing binary collisions with loss of energy due to normal and tangential restitution. In addition, we have simulated many particle systems of granular hard needles. The theory, based on the assumption of a homogeneous cooling state, predicts that granular cooling of the needles proceeds in two stages: An exponential decay of the initial configuration to a state where translational and rotational energies take on a time independent ratio (not necessarily unity), followed by an algebraic decay of the total kinetic energy $\sim t^{-2}$. The simulations support the theory very well for low and moderate densities. For higher densities, we have observed the onset of the formation of clusters and shear bands.Comment: 7 pages, 8 figures; major changes, extended versio

Spherically Symmetric Gravitational Collapse of General Fluids

We express Einstein's field equations for a spherically symmetric ball of general fluid such that they are conducive to an initial value problem. We show how the equations reduce to the Vaidya spacetime in a non-null coordinate frame, simply by designating specific equations of state. Furthermore, this reduces to the Schwarzschild spacetime when all matter variables vanish. We then describe the formulation of an initial value problem, whereby a general fluid ball with vacuum exterior is established on an initial spacelike slice. As the system evolves, the fluid ball collapses and emanates null radiation such that a region of Vaidya spacetime develops. Therefore, on any subsequent spacelike slice there exists three regions; general fluid, Vaidya and Schwarzschild, all expressed in a single coordinate patch with two free-boundaries determined by the equations. This implies complicated matching schemes are not required at the interfaces between the regions, instead, one simply requires the matter variables tend to the appropriate equations of state. We also show the reduction of the system of equations to the static cases, and show staticity necessarily implies zero ``heat flux''. Furthermore, the static equations include a generalization of the Tolman-Oppenheimer-Volkoff equations for hydrostatic equilibrium to include anisotropic stresses in general coordinates.Comment: 11 pages, 3 figures, submitted to Phys. Rev.

Sistem Informasi Strategis Mendayagunakan Sistem Informasi untuk Mencapai Keunggulan Kompetitif

While most companies are content to develop and implement computer-based information systems to improve their operational and managerial effectiveness, a few also rely on innovative systems to give them an edge over the competition. An information system that is specifically designed with the aim to create a competitive advantage for its firm is called a strategic information system.To build a strategic information system, one should have a good understanding as to what factors contribute towards competitive advantage. It is here that the analyses by Michael E. Porter on this subject are particularly invaluable. Furthermore, one should also realize what the modern computer excels at fast processing time, fast data retrieval, fast communications, and reliability.There are also risks inherent to strategic information systems. These include the large capital outlay involved, as well as the reality that any competitive advantage thus achieved is not sustainable for long

Is There Only One Solution of the Dyson-Schwinger Equation for Quark Propagator in the Case of Non-zero Current Quark Mass

In this letter it is shown on general ground that there exist two qualitatively distinct solutions of the Dyson-Schwinger equation for the quark propagator in the case of non-zero current quark mass. One solution corresponds to the ``Nambu-Goldstone'' phase and the other one corresponds to the ``Wigner'' phase in the chiral limit.Comment: 7 page

Dual neutral variables and knot solitons in triplet superconductors

In this paper we derive a dual presentation of free energy functional for spin-triplet superconductors in terms of gauge-invariant variables. The resulting equivalent model in ferromagnetic phase has a form of a version of the Faddeev model. This allows one in particular to conclude that spin-triplet superconductors allow formation of stable finite-length closed vortices (the knotted solitons).Comment: Replaced with version published in PRL (added a discussion of the effect of the coupling of the fields {\vec s} and {\vec C} on knot stability). Latest updates of the paper and miscellaneous links related to knotted solitons are also available at the homepage of the author http://www.teorfys.uu.se/PEOPLE/egor/ . Animations of knotted solitons by Hietarinta and Salo are available at http://users.utu.fi/h/hietarin/knots/c45_p2.mp