Electrical properties of the rabbit urinary bladder assessed using gramicidin D

Summary Recently, antibiotics have enjoyed widespread usage as tools in studies of epithelial transport. In the present study we assess the usefulness of the pore-forming antibiotic gramicidin D as a means for probing the electrical properties of the tight epithelium rabbit urinary bladder. Addition of 50 M gramicidin to the mucosal bath (either a NaCl or KCl Ringer's solution) led to a large irreversible increase in the transepithelial conductance (G _T ) within 800 sec.G _T increased by approximately 1200% and 500% in KCl and NaCl Ringer's solutions, respectively. Microelectrode measurements of the resistance ration (the ration of apical membrane resitance to basolateral membrane resistance) showed that apical membrane resistance is dereased by the drug. Measurements of the basolateral membrane resistance (R _bl ) and tight junctional resistance (R _j ) using a new and independent method (based on the perturbation of basolateral membrane electrogenic Na⁺ pump) demonstrated thatR _bl andR _j were unaffected, suggesting that the effects of gramicidin are restricted to the apical membrane for periods of at least 2 hours after drug addition. The selectivity of the gramicidin-induced permeability in the apical membrane was calculated from measurements of the apical membrane potential after ion substitutions using a modified version of the constant field equation. The selectivity sequence for cations was Cs⁺>K⁺>Na⁺>Li⁺>choline. Unlike the commonly used polyene antibiotics nystatin and amphotericin B, gramicidin did not induce a significant Cl^– permeability. In addition, the dose-response curve had a slope of 1. A method is described for calculating membrane resistances directly from transepithelial measurements under some conditions of gramicidin use, without requiring the use of microlectrode measurements.     

Sucrose and proton cotransport in Ricinus cotyledons

In Ricinus cotyledons, evidence for proton extrusion came from observation of direct acidification of the medium in the presence of potassium salts. Increasing K⁺ influx with increasing pH suggested a link between K⁺ influx and H⁺ efflux by an H⁺ pump. The kinetics of K⁺ influx and H⁺ efflux were consistent with a 1:1 stoichiometry K⁺:H⁺, which may indicate either electrical coupling or carrier mediated exchange. The results were consistent with an H⁺ pump setting up an electrochemical potential gradient which provides the driving force for an H⁺-sucrose cotransport and the movement of K⁺. With reference to this, a model for phloem loading is suggested.     

中脑导水管周围灰质在侧脑室注入微量吗啡或纳洛酮所引起的镇痛或拮抗镇痛中的作用

本工作进一步探索中脑导水管周围灰质(PAG)在吗啡镇痛与纳洛酮拮抗吗啡镇痛中的作用。实验在清醒受限制的大鼠上进行,以电刺激鼠尾出现的甩尾和嘶叫为痛反应指标。结果表明:(1)侧脑室注射微量纳洛酮后,可使电刺激 PAG 或注射微量吗啡于 PAG 所引起的镇痛效应受到明显拮抗;(2)损毀 PAG 或注射微量纳洛酮于 PAG 后,可使由侧脑室注入微量吗啡所引起的镇痛效应显著减弱。由此可见 PAG 既是侧脑室注射吗啡镇痛作用的重要中枢部位,又是侧脑室注射纳洛酮拮抗吗啡镇痛的重要中枢部位。     

Purification and reconstitution of the 32Pi-ATP exchange activity of bovine chromaffin granule membrane

Ghosts derived from bovine chromaffin granules have a ³²P_i-ATP exchange activity which is associated with the H⁺ pump of that membrane. This activity was low when compared to bacteria, chloroplasts or submitochondrial particles, but had similar properties (K_m for ATP and P_i, ATP/Mg²⁺ ratio, pH profile, inhibition by dicyclohexylcarbodiimide and tributyltin) to the ATPase from above membranes. The ³²P_i-ATP exchange activity was solubilized by cholate/octylglucoside mixtures. The soluble extract was lipid depleted by ammonium sulfate fractionation and partially purified by sucrose gradient centrifugation. The purified preparation was reconstituted with phospholipids by freeze-thawing. The reconstituted vesicles had a ³²P_i-ATP exchange sensitive to dicyclohexylcarbodiimide and trybutyltin and an ATPase with a sensitivity to the inhibitors which varied with the reconstitution conditions. The α- and β-subunits of F₁-ATPase were major components of the preparation.     

Cell fragility — The key problem of microalgae mass production in closed photobioreactors

The gap between the theoretical biological potential of microalgae and the biomass productivity obtained with algal culture in tubular biophotoreactors is due to a reduced growth rate related to hydrodynamic stress of pumping. High levels of mixing are necessary to reach a turbulent flow of the culture, in order to optimize the light regime. The optimal conditions of pumping to produce this significant liquid mixing may produce some cell damage. Factors affecting this hydrodynamic stress (geometry of the bioreactor involved, type of pump utilized, morphology of algal cells, physiological conditions of microalgae, etc.) are discussed.     

Current-voltage curve of electrogenic Cl− pump predicts voltage-dependent Cl− efflux inAcetabularia

Summary The current-voltage relationship of carrier-mediated, passive and active ion transport systems with one charge-carrying pathway can exactly be described by a simple reaction kinetic model. This model consists of two carrier states (one inside, one outside) and two pairs (forwards and backwards) of rate constants: a voltage-dependent one, describing the transport of charge and a voltage-insensitive one, summarizing all the other (voltage-independent) reactions. For the electrogenic Cl^– pump inAcetabularia these four rate constants have been determined from electrical measurements of the current-voltage relationship of the pump (Gradmann, Hansen & Slayman, 1981;in: Electrogenic Ion Pumps, Academic Press, New York). The unidirectional Cl^– efflux through the pump can also be calculated by the availiable reaction kinetic parameters.³⁶Cl^– efflux experiments on singleAcetabularia cells with simultaneous electrical stimulation (action potentials) and recording, demonstrate the unidirectional Cl^– efflux to depend on the membrane potential. After subtraction of an efflux portion which bypasses the pump, agreement is found between the measured flux-voltage relationship and the theoretical one as obtained from the reaction kinetic model and its parameters from the electrical data.     

The major growth current through lily pollen tubes enters as K+ and leaves as H+

Growing lily (Lilium longiflorum Thunb.) pollen always drive a current into their tubes and out of their grains. The only external ions needed for growth (and the growth current) are K⁺, H⁺, and Ca²⁺. Increases in K⁺ immediately stimulate the current; while decreases in K⁺ immediately inhibit it. Comparable changes in H⁺ have the opposite effect; while those in Ca²⁺ have very little effect. We infer that most of the steady growth current is carried in by a potassium leak and out by a proton pump; but other considerations indicate that a minor, but controlling, component of the inward current consists of calcium ions.     

Studies on normal human bone marrow cultures in controlled environment thin-film culture systems

Summary Two thin film culture systems, the controlled environment steady state system (SS) and the rocker tube configuration of that system (RT), were used to identify some of the conditions that appear to maintain morphologic and functional characteristics of cells of human bone marrow explants in vitro. The systems configuration assured continual gassing, control and easy monitoring of the cultures. Cytocentrifuge preparations of media of specimens cultured in RT disclosed, though in decreasing numbers, various hematopoietic cells for periods exceeding one month. Hematopoietic cells shed from specimens cultured in the SS system were retained in the culture tubes; cells of the myelocytic series predominated for the first 2 weeks while an increasing number of monocytes and macrophages appeared in the media of older cultures. Histologic examination of cultured explants disclosed preservation of the marrow architecture and the persistence of hematopoietic cells. Specimens cultured in RT tubes tended to be less cellular than similar cultures placed in dialysis bags or as cultured in the SS system. Immunoglobulins (Ig) were released into the culture media at a constant rate throughout the period of culture. Specimens that were cultured at a controlled pH of 7.4 released 2 to more than 4 times as much Ig as similar specimens maintained at a pH level of 7.1. There were no definitive differences in Ig levels in the cultures maintained at comparable pH levels and overlaid with various CO₂ concentrations, i.e. 2%, 5%, 10%; similarly, no differences in Ig levels were found in specimens cultured in media containing fetal bovine sera as opposed to horse sera. Supported by U.S.P.H.S. Grant CA-5834 from the National Cancer Institute. Department of Medicine A. Department of Cell Physiology Department of Immunology and Immunochemistry.     

Isolation of Thiomonas thermosulfatus Strain 51, a Species Capable of Coupling Biogenic Pyritization with Chemiosmotic Energy Transduction

The precipitation of iron sulfides potentially offers enough energy and reducing power to sustain life but organisms harnessing this reaction have not to our knowledge been previously described. We isolated a bacterial strain, capable of forming the iron sulfide minerals troilite (FeS), greigite (Fe₃S₄), and pyrite (FeS₂), from subsurface, microbial mats in Mangalia, Romania. This strain, most closely related to strains of Thiomonas sp., forms pyrite only if the redox conditions remain negative (< ?60 mV), sulfides are provided continually (≈1 mM), and the concentration of iron remains low (≤ 0.08 mM) but constant. Pyrite formation by this microbial strain is proposed as an example of biologically controlled mineralization because it is controlled by uncouplers of oxidative phosphorylation, it is larger in living than in dead cells, it is additive (controlled less by the amount of cell surfaces and more by reagents), and it results in the formation of ATP. This study indicates that precipitation and crystal formation can represent an energy resource for life and provides support for the “iron-sulfide world hypothesis” regarding the early evolution of life on Earth.