Specificity of siderophore receptors in membrane vesicles of Bacillus megaterium.

Membrane vesicles of Bacillus megaterium strains SK11 and Ard1 bound the ferrischizokinen and ferriferrioxamine B siderhores (iron transport cofactors). An approximately equimolar uptake of both labels of [3H, 59Fe]ferrischizokinen indicated binding of the intact chelate. Binding reached equilibrium in 2 to 5 min, was temperature independent, and was unaltered by the addition of several energy sources. A 91% dissociation of bound [Fe]ferrischizokinen was achieved in 60 s by the addition of excess ferrischizokinen. Ferriaerobactin, a siderophore which is structurally related to ferrischizokinen, caused no detectable release of bound [59Fe]ferrischizokinen. Of several other ferrigydroxamates tested, only ferriferrichrome A achieved the release (11%) of [Fe]ferrischizokinen. Rapid dissociation (92%) of bound [59Fe]ferriferrioxamine B by the addition of ferriferrioxamine B was observed, and a 67% release of [59Fe]ferriferrioxamine B was caused by ferriA2265, its structural relative. Ferrischizokinen, ferriferrichrome A, and ferrirhodotorulic acid produced a 6, 25, and 29% dissociation, respectively, of [59Fe]ferriferrioxamine B; ferriaerobactin caused no dissociation. [59Fe]ferriaerobactin was bound by the membranes, but its dissociation was not effected by unlabeled ferriaerobactin, suggesting no specific receptors for this chelate. The respective binding affinity constants and maximal binding capacities of membrane vesicles of strain SK11 were 2 x 10(7) M-1 and 280 pmol per mg of protein for ferrischizokinen and 7 x 10(7) M-1 and 37 pmol per mg of protein for ferriferrioxamine B. These values in strain Ard1 were, respectively, 1.4 x 10(7) M-1 and 186 pmol per mg of protein for ferrischizokinen and 11 x 10(7) M-1 and 23 pmol per mg of protein for ferriferrioxamine B. Separate, specific binding sites (receptors) for ferrischizokinen and ferriferrioxamine B exist on the vesicles. The ferrischizokinen receptors have a lower affinity but a higher binding capacity (eightfold) than that shown by the ferriferrioxamine B receptor. These receptors may be components of independent transport systems.     

Development,metamorphosis, and natural history of the nudibranch Doridella obscura Verrill (Corambidae: Opisthobranchia)

The doridacean nudibranch Doridella obscura Verrill was raised through one complete generation in laboratory culture, and spawning behavior monitored for a year at monthly intervals in Barnegat Bay, New Jersey.The nudibranch deposited egg masses throughout the year in Barnegat Bay, and the larvae remained viable at temperatures ranging from 1.5 to 28 °C. At 25 °C the eggs hatch 4 days after oviposition, and the planktotrophic veliger larvae swim and feed for 9 days before they metamorphose. Settlement occurs specifically on the bryozoan Electro crustulenta (Pallas). The spirally coiled larval shell grows rapidly until the dorsal mantle fold is retracted from the aperture 5–6 days after hatching. Although starved larvae grow only slightly and do not metamorphose, they resume normal development on introduction of suitable food. Newly metamorphosed juveniles consume algae and debris on the surface of the bryozoan until they grow large enough to attack the living zooids of E. crustulenta.The life cycle of Doridella obscura is short (26 days at 25 °C), allowing the nudibranchs to take advantage of short-lived Electra crustulenta colonies in unstable habitats in bays and estuaries.     

Microbial metabolism of amino alcohols. Control of formation and stability of partially purified ethanolamine ammonia-lyase in Escherichia coli.

Induction of ethanolamine ammonia-lyase formation in Escherichia coli required both the ethanolamine and vitamin B12, and was gratuitous during growth on glycerol. Ethanolamine analogues inhibited enzyme activity and inhibited growth with ethanolamine as the the nitrogen source, but did not act as inducers. Enzyme formation was more rapid when ethanolamine was added to cultures containing vitamin B12 rather than the reverse. Enzyme formation was subject to catabolic repression, glucose and acetate being particularly effective. Chloramphenicol, I-aminopropan 2-01 and 1,3-diaminopropan-2-01 prevented enzyme induction. Ethanolamine ammonia-lyase, resolved from its cobamide coenzyme, was purified 35-fold. The apoenzyme was stable for several days in the presence of ethanolamine, dithiothreitol, glycerol and K+ ions. Enzyme formation therefore requires both substrate and cobamide coenzyme to be present simultaneously as inducers.     

Cytomat-R: a computer-controlled multiple laser source multiparameter flow cytophotometer system.

A multiple illumination wavelength multiparameter flow cytophotometer system, using laser sources and controlled by a small, general-purpose digital computer, has been produced for use in the development of new flow cytometric techniques. Three different laser wave-lengths can be used simultaneously to illuminate different regions of the flow chamber; as many as five measurements of light scattering at various angles, extinction, and fluorescence at one or more wavelengths can be made at each illuminated station. Cells in suspension may be examined at rates of 1000 cells/sec, with seven correlated optical measurements being recorded for each cell. A library of programs for data manipulation and statistical analysis make it possible to use the system to develop and implement cell characterization, counting and classification procedures for basic and clinical research applications.     

Cell-specific differences in membrane beta-glucosidase from normal and Gaucher cells.

Two isozymes of membrane-bound beta-glucosidase (beta-D-glucoside glucohydrolase, EC 3.2.1.21) with activity towards 4-methylumbelliferyl-beta-D-glucopyranoside have been identified in human cells. One of these isozymes was found to have a pH optimum of 5.0, a Km of 0.4 mM and to be rapidly inactivated at pH 4.0 ("acid-labile"). The second isozyme had a pH optimum of 4.5, a Km of 0.8 mM and was stable at pH 4.0 ("acid-stable"). Cultured long-term lymphoid lines and peripheral blood leukocytes contained both isozymes while cultured skin fibroblasts contained only the "acid-stable" form in detectable amounts. The specific activity of the "acid-stable" isozyme was severely reduced in cultured skin fibroblasts, cultured long-term lines and peripheral leukocytes from patients with Gaucher's disease. The specific activity of the "acid-labile" enzyme in the latter two cell types was apparently unaffected. The beta-glucosidase activity in all three cell types examined was predominantly particulate but the enzyme could be solubilized with low concentrations of Triton X-100. The solubilized enzyme required sodium taurocholate (0.2%) for maximum activity. Solubilized beta-glucosidase did not exhibit the cell-specific differences in pH optimum and Km shown by the membrane-bound enzyme.     

Further studies of proximal tubular brush border membraned-glucose transport heterogeneity

Summary The properties of two sodium-dependentd-glucose transporters previously identified in renal proximal tubule brush border membrane (BBM) vesicles are studied. The low-affinity system, found in BBM vesicles from the outer cortex (early proximal tubule), is shown to be associated with the high-affinity phlorizin binding site typically found in renal BBM preparations. The high-affinity system, found in BBM vesicles from the outer medulla (late proximal tubule), is almost two orders of magnitude less sensitive to inhibition by phlorizin and is apparently not associated with high-affinity phlorizin binding. The sodium/g;ucose stoichiometry of the outer medullary transporter is found to be 21 by two independent methods. Previous measurements have established that the stoichiometry of the outer cortical system is 11. It is suggested that this arrangement of transporters in series along the proximal tubule enables the kidney to reabsorb glucose from the urine in an energy-efficient fashion. The bulk of the glucose load is reabsorbed early in the proximal tubule at an energetic cost of one Na⁺ per glucose molecule. Then in the late proximal tubule a larger coupling ratio and hence a larger driving force is employed to reabsorb the last traces of glucose from the urine.     

Hepatocystis Parasitemia in Wild Kenya Vervet Monkeys (Cercopithecus aethiops)

Blood smears of 159 vervet monkeys from three sites in Kenya were stained with Giemsa and examined for Hepatocystis parasites. The populations differ in incidence of parasitemia, ranging from 0–62% affected individuals. These differences are probably due to altitude and local environmental conditions.     

Cycling of organic and inorganic sulphur in a chestnut oak forest

Summary Sulfur (S) cycling in a chestnut oak forest on Walker Branch Watershed, Tennessee, was dominated by geochemical processes involving sulfate. Even though available SO ₄ ^2- was present far in excess of forest nutritional requirements, the ecosystem as a whole accumulated 60% of incoming SO₄–S. Most (90%) of this accumulation occurred by SO ₄ ^2- adsorption in sesquioxide-rich subsurface soils, with a relatively minor amount accumulating and cycling as SO ₄ ^2- within vegetative components. Organic sulfates are thought to constitute a large proportion of total S in surface soils, also, and to provide a pool of readily mineralized available S within the ecosystem.Research sponsored by Division of Biomedical and Environmental Research, U.S. Department of Energy, under contract W-7405-eng-26 with Union Carbide Corporation. Soil ester sulfate work sponsored by contract RP-1813-1 with the Electric Power Research Institute. Publication No. 1990, Environmental Sciences Division, Oak Ridge National Laboratory, Oak Ridge, Tennessee 37830     

Persistent atrial paralysis: Case report with light microscopy and ultrastructural analyses

Persistent atrial paralysis in a patient with complete heart block and mild mitral insufficiency is presented. Left atrial specimens obtained during implantation of a permanent cardiac pulse generator showed evidence of hypertrophy and fibrosis; subcellular degenerative changes ranged from near normal to irreversible, thus suggesting that atrial paralysis may be due to the replacement of normal atrial muscle with nonfunctional fibrous tissue.     

Phosphatidylethanolamine distribution and fluidity in outer and inner membranes of the gram-negative bacterium Erwinia carotovora.

1. The distribution of phosphatidylethanolamine, the major lipid of Erwinia carotovora, was investigated in intact bacteria, spheroplasts and outer- and inner-membrane preparations, with the amino-group reagent 2,4,6-trinitrobenzenesulphonic acid. Only 4% was found on the external surface of the outer membrane with 30% on the internal surface, whereas the inner membrane had 27 and 38% on its external and internal surfaces respectively. Some comparative studies were made with three other bacteria. 2. The fluidity of the membranes of E. carotovora was studied by using the fluorescent probe 1,6-diphenylhexa-1,3,5-triene. Results were consistent with the hydrocarbon region of the outer membrane bilayer being less fluid than that of the inner one. 3. On the basis of these and other results a model for the outer- and inner-membrane structures of E. carotovora is proposed.