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

Gas Requirements in Pressurized Transfer of Liquid Hydrogen

Of late, liquid hydrogen has become a very popular fuel for space missions. It is being used in such programs as Centaur and Saturn. Furthermore, hydrogen is the ideal working fluid for nuclear powered space vehicles currently under development. In these applications, liquid hydrogen fuel is generally transferred to the combustion chamber by a combination of pumping and pressurization. The pump forces the liquid propellant from the fuel tank to the combustion chamber; gaseous pressurant holds tank pressure sufficiently high to prevent cavitation at the pump inlet and to maintain the structural rigidity of the tank. The pressurizing system, composed of pressurant, tankage, and associated hardware can be a large portion of the total vehicle weight. Pressurant weight can be reduced by introducing the pressurizing gas at temperatures substantially greater than those of liquid hydrogen. Heat and mass transfer processes thereby induced complicate gas requirements during discharge. These requirements must be known to insure proper design of the pressurizing system. The aim of this paper is to develop from basic mass and energy transfer processes a general method to predict helium and hydrogen gas usage for the pressurized transfer of liquid hydrogen. This required an analytical and experimental investigation, the results of which are described in this paper

Apical endosomes isolated from kidney collecting duct principal cells lack subunits of the proton pumping ATPase.

Endocytic vesicles that are involved in the vasopressin-stimulated recycling of water channels to and from the apical membrane of kidney collecting duct principal cells were isolated from rat renal papilla by differential and Percoll density gradient centrifugation. Fluorescence quenching measurements showed that the isolated vesicles maintained a high, HgCl2-sensitive water permeability, consistent with the presence of vasopressin-sensitive water channels. They did not, however, exhibit ATP-dependent luminal acidification, nor any N-ethylmaleimide-sensitive ATPase activity, properties that are characteristic of most acidic endosomal compartments. Western blotting with specific antibodies showed that the 31- and 70-kD cytoplasmically oriented subunits of the vacuolar proton pump were not detectable in these apical endosomes from the papilla, whereas they were present in endosomes prepared in parallel from the cortex. In contrast, the 56-kD subunit of the proton pump was abundant in papillary endosomes, and was localized at the apical pole of principal cells by immunocytochemistry. Finally, an antibody that recognizes the 16-kD transmembrane subunit of oat tonoplast ATPase cross-reacted with a distinct 16-kD band in cortical endosomes, but no 16-kD band was detectable in endosomes from the papilla. This antibody also recognized a 16-kD band in affinity-purified H+ ATPase preparations from bovine kidney medulla. Therefore, early endosomes derived from the apical plasma membrane of collecting duct principal cells fail to acidify because they lack functionally important subunits of a vacuolar-type proton pumping ATPase, including the 16-kD transmembrane domain that serves as the proton-conducting channel, and the 70-kD cytoplasmic subunit that contains the ATPase catalytic site. This specialized, non-acidic early endosomal compartment appears to be involved primarily in the hormonally induced recycling of water channels to and from the apical plasma membrane of vasopressin-sensitive cells in the kidney collecting duct

The Kinetic Interpretation of the DGLAP Equation, its Kramers-Moyal Expansion and Positivity of Helicity Distributions

According to a rederivation - due to Collins and Qiu - the DGLAP equation can be reinterpreted (in leading order) in a probabilistic way. This form of the equation has been used indirectly to prove the bound $|\Delta f(x,Q)| < f(x,Q)$ between polarized and unpolarized distributions, or positivity of the helicity distributions, for any $Q$. We reanalize this issue by performing a detailed numerical study of the positivity bounds of the helicity distributions. To obtain the numerical solution we implement an x-space based algorithm for polarized and unpolarized distributions to next-to-leading order in $\alpha_s$, which we illustrate. We also elaborate on some of the formal properties of the Collins-Qiu form and comment on the underlying regularization, introduce a Kramers-Moyal expansion of the equation and briefly analize its Fokker-Planck approximation. These follow quite naturally once the master version is given. We illustrate this expansion both for the valence quark distribution $q_V$ and for the transverse spin distribution $h_1$.Comment: 38 pages, 27 figures, Dedicated to Prof. Pierre Ramond for his 60th birthda

A preliminary assessment of the geologic setting, hydrology, and geochemistry of the Hueco Tanks geothermal area, Texas and New Mexico

Tx Doc no. : Z, UA220.7, G292, 81-1UT Librarie

Shadowing Effects on the Nuclear Suppression Factor, R_dAu, in d+Au Interactions

We explore how nuclear modifications to the nucleon parton distributions affect production of high transverse momentum hadrons in deuteron-nucleus collisions. We calculate the charged hadron spectra to leading order using standard fragmentation functions and shadowing parameterizations. We obtain the d+Au to pp ratio both in minimum bias collisions and as a function of centrality. The minimum bias results agree reasonably well with the BRAHMS data while the calculated centrality dependence underestimates the data and is a stronger function of p_T than the data indicate.Comment: 18 pages, 3 figures, final version, Phys. Rev. C in pres

Polarized parton distributions from NLO QCD analysis of world DIS and SIDIS data

The combined analysis of polarized DIS and SIDIS data is performed in NLO QCD. The new parametrization on polarized PDFs is constructed. The uncertainties on PDFs and their first moments are estimated applying the modified Hessian method. The especial attention is paid to the impact of novel SIDIS data on the polarized distributions of light sea and strange quarks. In particular, the important question of polarized sea symmetry is studied in comparison with the latest results on this subject