Structure of the novel membrane-coating material in proton-secreting epithelial cells and identification as an H+ATPase.

Specialized proton-secreting cells known collectively as mitochondria-rich cells are found in a variety of transporting epithelia, including the kidney collecting duct (intercalated cells) and toad and turtle urinary bladders. These cells contain a population of characteristic tubulovesicles that are believed to be involved in the shuttling of proton pumps (H+ATPase) to and from the plasma membrane. These transporting vesicles have a dense, studlike material coating the cytoplasmic face of their limiting membranes and similar studs are also found beneath parts of the plasma membrane. We have recently shown that this membrane coat does not contain clathrin. The present study was performed to determine the structure of this coat in rapidly frozen and freeze-dried tissue, and to determine whether the coat contains a major membrane protein transported by these vesicles, a proton pumping H+ATPase. The structure of the coat was examined in proton-secreting, mitochondria-rich cells from toad urinary bladder epithelium by rapidly freezing portions of apical membrane and associated cytoplasm that were sheared away from the remainder of the cell using polylysine-coated coverslips. Regions of the underside of these apical membranes as large as 0.2 micron2 were decorated by studlike projections that were arranged into regular hexagonal arrays. Individual studs had a diameter of 9.5 nm and appeared to be composed of multiple subunits arranged around a central depression, possibly representing a channel. The studs had a density of approximately 16,800 per micron2 of membrane. Similar arrays of studs were also found on vesicles trapped in the residual band of cytoplasm that remained attached to the underside of the plasma membrane, but none were seen in adjacent granular cells. To determine whether these arrays of studs contained H+ATPase molecules, we examined a preparation of affinity-purified bovine medullary H+ATPase, using the same technique, after incorporation of the protein eluted from a monoclonal antibody affinity column into phospholipid liposomes. The affinity-purified protein was shown to be capable of ATP-dependent acidification. In such preparations, large paracrystalline arrays of studs identical in appearance to those seen in situ were found. The dimensions of the studs as well as the number per square micrometer of membrane were identical to those of toad bladder mitochondria-rich cells: 9.5 nm in diameter, 16,770 per micron2 of membrane.(ABSTRACT TRUNCATED AT 400 WORDS

A New Approach to Black Hole Microstates

If one encodes the gravitational degrees of freedom in an orthonormal frame field there is a very natural first order action one can write down (which in four dimensions is known as the Goldberg action). In this essay we will show that this action contains a boundary action for certain microscopic degrees of freedom living at the horizon of a black hole, and argue that these degrees of freedom hold great promise for explaining the microstates responsible for black hole entropy, in any number of spacetime dimensions. This approach faces many interesting challenges, both technical and conceptual.Comment: 6 pages, 0 figures, LaTeX; submitted to Mod. Phys. Lett. A.; this essay received "honorable mention" from the Gravity Research Foundation, 199

Action and Energy of the Gravitational Field

We present a detailed examination of the variational principle for metric general relativity as applied to a ``quasilocal'' spacetime region \M (that is, a region that is both spatially and temporally bounded). Our analysis relies on the Hamiltonian formulation of general relativity, and thereby assumes a foliation of \M into spacelike hypersurfaces $\Sigma$. We allow for near complete generality in the choice of foliation. Using a field--theoretic generalization of Hamilton--Jacobi theory, we define the quasilocal stress-energy-momentum of the gravitational field by varying the action with respect to the metric on the boundary \partial\M. The gravitational stress-energy-momentum is defined for a two--surface $B$ spanned by a spacelike hypersurface in spacetime. We examine the behavior of the gravitational stress-energy-momentum under boosts of the spanning hypersurface. The boost relations are derived from the geometrical and invariance properties of the gravitational action and Hamiltonian. Finally, we present several new examples of quasilocal energy--momentum, including a novel discussion of quasilocal energy--momentum in the large-sphere limit towards spatial infinity.Comment: To be published in Annals of Physics. This final version includes two new sections, one giving examples of quasilocal energy and the other containing a discussion of energy at spatial infinity. References have been added to papers by Bose and Dadhich, Anco and Tun

Hamiltonians for a general dilaton gravity theory on a spacetime with a non-orthogonal, timelike or spacelike outer boundary

A generalization of two recently proposed general relativity Hamiltonians, to the case of a general (d+1)-dimensional dilaton gravity theory in a manifold with a timelike or spacelike outer boundary, is presented.Comment: 17 pages, 3 figures. Typos correcte

TMS communications software. Volume 1: Computer interfaces

A prototype bus communications system, which is being used to support the Trend Monitoring System (TMS) as well as for evaluation of the bus concept is considered. Hardware and software interfaces to the MODCOMP and NOVA minicomputers are included. The system software required to drive the interfaces in each TMS computer is described. Documentation of other software for bus statistics monitoring and for transferring files across the bus is also included

Canonical Quasilocal Energy and Small Spheres

Consider the definition E of quasilocal energy stemming from the Hamilton-Jacobi method as applied to the canonical form of the gravitational action. We examine E in the standard "small-sphere limit," first considered by Horowitz and Schmidt in their examination of Hawking's quasilocal mass. By the term "small sphere" we mean a cut S(r), level in an affine radius r, of the lightcone belonging to a generic spacetime point. As a power series in r, we compute the energy E of the gravitational and matter fields on a spacelike hypersurface spanning S(r). Much of our analysis concerns conceptual and technical issues associated with assigning the zero-point of the energy. For the small-sphere limit, we argue that the correct zero-point is obtained via a "lightcone reference," which stems from a certain isometric embedding of S(r) into a genuine lightcone of Minkowski spacetime. Choosing this zero-point, we find agreement with Hawking's quasilocal mass expression, up to and including the first non-trivial order in the affine radius. The vacuum limit relates the quasilocal energy directly to the Bel-Robinson tensor.Comment: revtex, 22 p, uses amssymb option (can be removed

Diagnostic procedures for Trend Monitoring System (TMS) communications

A prototype coaxial cable bus communications sytem was developed to support the trend monitoring system (TMS). Troubleshooting procedures are described at the system level. The procedures can be used by repair personnel to isolate a fault in the TMS and to restore the system to operation by swapping out failed components

Putting an Edge to the Poisson Bracket

We consider a general formalism for treating a Hamiltonian (canonical) field theory with a spatial boundary. In this formalism essentially all functionals are differentiable from the very beginning and hence no improvement terms are needed. We introduce a new Poisson bracket which differs from the usual ``bulk'' Poisson bracket with a boundary term and show that the Jacobi identity is satisfied. The result is geometrized on an abstract world volume manifold. The method is suitable for studying systems with a spatial edge like the ones often considered in Chern-Simons theory and General Relativity. Finally, we discuss how the boundary terms may be related to the time ordering when quantizing.Comment: 36 pages, LaTeX. v2: A manifest formulation of the Poisson bracket and some examples are added, corrected a claim in Appendix C, added an Appendix F and a reference. v3: Some comments and references adde

Motion and Trajectories of Particles Around Three-Dimensional Black Holes

The motion of relativistic particles around three dimensional black holes following the Hamilton-Jacobi formalism is studied. It follows that the Hamilton-Jacobi equation can be separated and reduced to quadratures in analogy with the four dimensional case. It is shown that: a) particles are trapped by the black hole independently of their energy and angular momentum, b) matter alway falls to the centre of the black hole and cannot understake a motion with stables orbits as in four dimensions. For the extreme values of the angular momentum of the black hole, we were able to find exact solutions of the equations of motion and trajectories of a test particle.Comment: Plain TeX, 9pp, IPNO-TH 93/06, DFTUZ 93/0

Boundary States and Black Hole Entropy

Black hole entropy is derived from a sum over boundary states. The boundary states are labeled by energy and momentum surface densities, and parametrized by the boundary metric. The sum over state labels is expressed as a functional integral with measure determined by the density of states. The sum over metrics is expressed as a functional integral with measure determined by the universal expression for the inverse temperature gradient at the horizon. The analysis applies to any stationary, nonextreme black hole in any theory of gravitational and matter fields.Comment: 4 pages, Revte