Parameters of gaseous ion effects on the mammalian trachea. A. Duration of effects. B. Minimal effective ion densities

A. Duration of Effects Groups of mice exposed to high densities of unipolar light air ions for 72 hours exhibited persistent alterations in the functional efficiency of their tracheas. These effects lasted at least 4 weeks, and in the case of animals treated with (+) ions included diminished ciliary activity, pale and contracted tracheal mucosa, and enhanced vulnerability to trauma. Following treatment with (-) ions, animals displayed increased ciliary activity with no other detectable changes. It required at least 60 minutes of exposure to ions to induce such "permanent" functional changes. B. Minimal Effective Ion Densities The minimal ion densities producing changes in ciliary activity within an arbitrary period of 30 minutes were determined with extirpated tracheal strips from rabbits and guinea pigs. The threshold value for (-) ions was approximately 2.5 x 10³ ions/cm.²/sec. and that for (+) ions was in the range between 1 x 10⁴ and 2.5 x 10⁵ ions/cm.²/sec.The minimal ion densities producing changes in ciliary activity within an arbitrary period of 30 minutes were determined with extirpated tracheal strips from rabbits and guinea pigs. The threshold value for (-) ions was approximately 2.5 x 10³ ions/cm.²/sec. and that for (+) ions was in the range between 1 x 10⁴ and 2.5 x 10⁵ ions/cm.²/sec. The evidence indicates that ion-induced functional changes in the ciliated epithelium of the pulmonary tree are the results of direct contact of ions with surface cells and do not involve participation of the central nervous system or circulation. So far as ciliary activity is concerned, the number of ions required to produce a change in rate is very small.     

Isolation and biological activity of the proteases released by sea urchin eggs following fertilization.

The protease activity released from sea urchin egg cortical granules into the surrounding seawater at fertilization is involved in vitelline layer elevation and the block to polyspermy. The cortical granule protease components were isolated by isoelectric precipitation and affinity chromatography on p-aminobenzamidine-Sepharose columns. Elution profiles from affinity columns suggested heterogeneity of the proteases, and polyacrylamide-gel electrofocusing of affinity-purified preparations established the presence of two proteins. Dramatically different biological activities were resolved by affinity chromatography. Early-eluting fractions of low specific activity delaminated the vitelline layer from the egg plasma membrane; this activity is termed vitelline delaminase. Late-eluting fractions of high specific activity modified the egg vitelline layer surface such that sperm could not bind or fertilize them; this activity is referred to as sperm receptor hydrolase. The biological activities of the sea urchin proteases are apparently the result of limited action on the vitelline layer, unlike bovine trypsin which simply digests the vitelline layer. The cortical granule proteases lost biological specificity when stored at 0°C at pH 8.0. Esterase activity increased, and the preparation acquired the ability to digest the vitelline layer. Increase of the esterase activity in protease preparations was prevented by storage at low pH.The molecular weight of both enzymes was estimated by sucrose gradient centrifugation to be 47,000, whereas multiple components with molecular weights between 10⁵ and 10⁶ were demonstrated by gel filtration.     

Enhancement of evaporation rates from thin layers of liquids exposed to air ions

A beta-ray gauge system was used to study evaporation rates and drying times of liquids exposed to relatively high fluxes of air ions of either polarity produced by a corona discharge. Evaporation rates from thin layers of water, ethyl alcohol, and carbon tetrachloride increased considerably when exposed to air ions, compared to untreated liquids under constant environmental conditions. Drying times of liquid layers exposed to air ions were shortened by a factor of at least two over liquids that had not been exposed to ions. The drying time of a liquid layer was found to increase with the decrease of ion fluxes at the liquid surface. Threshold values of 1.9×10¹¹ positive ions and 2.7×10¹¹ negative ions cm^–2 s^–1 were established below which no increase in the evaporation rates were observed for ethyl alcohol and carbon tetrachloride. The evaporation rate of water at these same ion fluxes was significantly higher than that of the control. The present experiments indicate that a stream of air ions could play a major role in the observed enhancement of evaporation rates.     

Continuous production of lignin peroxidase by immobilized Phanerochaete chrysosporium in a pilot scale bioreactor

Lignin peroxidase was continuously produced by nylon-web or polyurethane immobilized Phanerochaete chrysosporium ATCC 24725 in a modified Biostate E® bioreactor, agitated either with the air and oxygen flow alone or in combination with a mechanical stirrer. Lignin peroxidase production started rapidly, and activities as high as ∼600 U l⁻¹ were reached as early as in 3 days. At best a total activity yield of ∼10 000 U was obtained during one week's continuous production with the maximum activity of ∼750 U l⁻¹ with nylon-web immobilized fungus, veratryl alcohol as an activator, and 2,2-dimethylsuccinate as buffer, although the relatively inexpensive benzyl alcohol and sodium tartrate peformed satisfactorily as the activator and buffer, respectively.     

The distribution of tetramethylammonium ions on the surface of purple membranes

The two-dimensional distribution of deuterated tetramethylammonium (TMA⁺) ions on the surface of purple membranes of Halobacterium halobium was determined by neutron diffraction. The measurements were performed on stacks of these membranes with a high concentration of TMA⁺ molecules in the water layer between the membranes. A difference Fourier analysis of samples with deuterated and undeuterated ions showed an excess of 8.5 TMA⁺ ions per elementary cell in the lipid areas compared to the protein areas. A total number of 90 ions per elementary cell in the intermembrane space was estimated from the preparation procedure. The excess in the lipid domains may result from the higher affinity of TMA⁺ ions for the lipid head groups and/or from the fact that the protein (bacteriorhodopsin) protrudes slightly out of the lipid surface.Abbreviations BR bacteriorhodopsin - TMA Tetramethylammonium     

Dielectric saturation of the aqueous boundary layers adjacent to charged bilayer membranes

Summary The surface charge density resulting from the adsorption of hydrophobic anions of dipicrylamine onto dioleyl-lecithin bilayer membranes has been measured directly using a high field pulse method. The surface charge density increases linearly with adsorbate concentration in the water until electrostatic repulsion of impinging hydrophobic ions by those already adsorbed becomes appreciable. Then Gouy-Chapman theory predicts that surface charge density will increase sublinearly, with the power [z ⁺/(z ⁺+2)] of the adsorbate concentration, wherez ⁺ is the cation valence of the indifferent electrolyte screening the negatively charged membrane surface. The predicted 1/3 and 1/2 power laws for univalent and divalent cations, respectively, have been observed in these experiments using Na⁺, Mg⁺⁺, and Ba⁺⁺ ions. Gouy-Chapman theory predicts further that the change from linear to sublinear dependence takes place at a surface charge density governed by the static dielectric constant of water and the concentration of indifferent electrolyte. Quantitative agreement with experiment is obtained at electrolyte concentrations of 10^–4 m and 10^–3 m, but can be maintained at higher concentrations only if the aqueous dielectric constant is decreased. A transition field model is proposed in which the Gouy-Chapman theory is modified to take account of dielectric saturation of water in the intense electric fields adjacent to charged membrane surfaces.     

In Situ Generation of Artificial Solid‐Electrolyte Interphases on 3D Conducting Scaffolds for High‐Performance Lithium‐Metal Anodes

Rational structure design of the current collector along with further engineering of the solid‐electrolyte interphases (SEI) layer is one of the most promising strategies to achieve uniform Li deposition and inhibit uncontrolled growth of Li dendrites. Here, a Li₂S layer as an artificial SEI with high compositional uniformity and high lithium ion conductivity is in situ generated on the surface of the 3D porous Cu current collector to regulate homogeneous Li plating/stripping. Both simulations and experiments demonstrate that the Li₂S protective layer can passivate the porous Cu skeleton and balance the transport rate of lithium ions and electrons, thereby alleviating the agglomerated Li deposition at the top of the electrode or at the defect area of the SEI layer. As a result, the modified current collector exhibits long‐term cycling of 500 cycles at 1 mA cm^?2 and stable electrodeposition capabilities of 4 mAh cm^?2 at an ultrahigh current density of 4 mA cm^?2. Furthermore, full batteries (LiFePO₄ as cathode) paired with this designed 3D anode with only ≈200% extra lithium show superior stability and rate performance than the batteries paired with lithium foil (≈3000% extra lithium). These explorations provide new strategies for developing high‐performance Li metal anodes.     

Anchorage-dependent mammalian cell culture using polyurethane foam as a new substratum for cell attachment

Summary Anchorage-dependent mammalian cells were cultivated at high cell density in a novel culture system using polyurethane foam (PUF) as a substratum for cell attachment. PUF has a macroporous structure giving a high surface area to volume ratio. Monkey kidney cells (Vero) and Chinese hamster ovary cells (CHO-K1) attached to the internal surface of PUF and grew to a high cell density (1.04 × 10⁸ cells/ cm³ PUF and 3.5 × 10⁷ cells/ cm³ PUF, respectively) in PUF stationary cultures. In addition, we have designed a PUF-particle packed-bed culture system for high density mass cell culture. A maximum cell density of 2.4 × 10⁷ cells/cm³ culture vessel volume was obtained in a packed-bed culture of Vero cells. Offprint requests to: K. Funatsu     

Surface potential of phosphatidylserine monolayers. I. Divalent ion binding effect

Surface potentials of phosphatidylserine monolayers have been measured in the presence of different divalent ion concentrations in order to determine the way in which divalent ions bind to the membrane surface. The association constants for divalent ions (Mg²⁺, Ca²⁺ and Mn²⁺) with the phosphatidylserine membrane have been obtained from the experimental data and simple ion binding theory. The order of divalent ion binding to the membrane is Mn²⁺ > Ca²⁺ > Mg²⁺. However, none of the divalent ions used completely neutralized the negative charge of phosphatidylserine even at relatively high concentrations. The amounts of the divalent ions bound depended upon the concentration of the monovalent ions present in the subphase. It is suggested that the amounts of bound ions obtained from the use of radioisotope tracer methods may include a considerable contribution from the excess free ions in the double layer region of the phosphatidylserine membrane.     

Spatio-Temporal Distributions of Metal Ions and Taxol of Taxus cuspidata Cells Immobilized on Polyurethane Foam

Distinct spatio-temporal variations of metal ions and Taxol production were observed for Taxus cuspidata cells immobilized on polyurethane foam. The Taxol content in the inner foam layer reached 215 μg g⁻¹ at day 30, which was 40-fold higher than that in the outer foam layer, and the Ca²⁺ and Mg²⁺ contents were 5.3 and 3.7 times higher, while the K⁺ content was 5.5 times lower. Thus higher intracellular Ca²⁺ and Mg²⁺ contents and lower intracellular K⁺ content may favor the Taxol biosynthesis in immobilized Taxus cuspidata.     

Effect of Heavy Metal Ions on the Growth and Iron-oxidizing Activity of Thiobacillus ferrooxidans

Effect of heavy metal ions on the growth and the iron-oxidizing activity of Thiobacillus ferrooxidans were investigated.

Cupric, zinc, cadmium, and chromium ions had no effect on the growth and the iron-oxidizing activity of cell suspensions or cell-free extracts of the bacterium in high concentrations (10^?3~10^?2M). Lead ion delayed the start of the growth slightly in 10^?3 M, but it did not inhibit the iron-oxidizing activity of the cells in the concentration. Tin and molybdenum oxide ions inhibited both of them in the concentration above 10^?3 M.

Mercuric mercurous, and silver ions had the most harmful effect. In the concentration of 10^?3 .M, each of the cations inhibited almost completely both the growth and the iron-oxidizing activity of the cells.

In the experiments with cell-free extracts it was observed that the activity of cytochrome oxidase (cytochrome a₅₉₇) operating in the iron-oxidizing system of the bacterium was specifically inhibited with mercuric ion in the concentration above 5 × 10^?4 M.     

Surface potential effects on metal ion binding to phosphatidylcholine membranes. 31P NMR study of lanthanide and calcium ion binding to egg-yolk lecithin vesicles

³¹P NMR of phosphatidylcholine (lecithin) from egg-yolk in sonicated vesicles has been measured in the presence of various ions. Addition of Ln³⁺³ or Ca²⁺ shifted the ³¹P resonance of the phosphate groups of the outer surface of the vesicles. These shifts were measured at varied lanthanide or Ca²⁺ concentration at different ionic strengths obtained by addition of NaCl. The shifts induced by Tb³⁺ and Ca²⁺ have been analyzed using the theory of the diffuse double layer. Corrections were introduced for the effect of the ionic strength on the activities of the ions. The binding efficiency is shown to be controlled by the electrostatic potential produced by the bound cations at the membrane surface. This potential is slightly modified due to weak chloride binding. Binding constants have been derived.     

Superior Sodium Storage in Na2Ti3O7 Nanotube Arrays through Surface Engineering

Sodium‐ion batteries have attracted extraordinary attention owing to their low cost and raw materials in abundance. A major challenge of practical implementation is the lack of accessible and affordable anodes that can reversibly store a substantial amount of Na ions in a fast and stable manner. It is reported that surface engineered sodium titanate (Na₂Ti₃O₇) nanotube arrays directly grown on Ti substrates can serve as efficient anodes to meet those stringent requirements. The fabrication of the nanotube arrays involves hydrothermal growing of Na₂Ti₃O₇ nanotubes, surface deposition of a thin layer of TiO₂, and subsequent sulfidation. The resulting nanoarrays exhibit a high electrochemical Na‐storage activity that outperforms other Na₂Ti₃O₇ based materials. They deliver high reversible capacities of 221 mAh g^?1 and exhibit a superior cycling efficiency and rate capability, retaining 78 mAh g^?1 at 10 C (1770 mA g^?1) over 10 000 continuous cycles. In addition, the full cell consisting of Na₂Ti₃O₇ nanotube anode and Na_2/3(Ni_1/3Mn_2/3)O₂ cathode is capable of delivering a specific energy of ≈110 Wh kg^?1 (based on the mass of both electrodes). The surface engineering can provide useful tools in the development of high performance anode materials with robust power and cyclability.     

Microbially Induced Calcium Carbonate Precipitation on Surface or in the Bulk of Soil

Microbial precipitation of calcium carbonate takes place in nature by different mechanisms. One of them is microbially induced carbonate precipitation (MICP), which is performed due to bacterial hydrolysis of urea in soil in the presence of calcium ions. The MICP process can be adopted to reduce the permeability and/or increase the shear strength of soil. In this paper, a study on the use of Bacillus sp., which was isolated from tropical beach sand, to perform MICP either on the surface or in the bulk of sand is presented. If the level of calcium salt solution was below the sand surface, MICP took place in the bulk of sand. On the other hand, if the level of calcium salt solution was above the sand surface, MICP was performed on the sand surface and formed a thin layer of crust of calcium carbonate. After six sequential batch treatments with suspension of urease-producing bacteria and solutions of urea and calcium salt, the permeability of sand was reduced to 14 mm/day (or 1.6×10^?7 m/s) in both cases of bulk and surface MICP. Quantities of precipitated calcium after six treatments were 0.15 and 0.60 g of Ca per cm² of treated sand surface for the cases of bulk or surface MICP, respectively. The stiffness of the MICP treated sand also increased considerably. The modulus of rupture of the thin layer of crust was 35.9 MPa which is comparable with limestone.     

Temporal Variation of Denitrification Activity in Plant-Covered, Littoral Sediment from Lake Hampen, Denmark

Diel and seasonal variations in denitrification were determined in a littoral lake sediment colonized by the perennial macrophyte Littorella uniflora (L.) Aschers. In the winter, the activity was low (5 μmol of N m⁻² h⁻¹) and was restricted to the uppermost debris layer at a depth of 0 to 1 cm. By midsummer, the activity increased to 50 μmol of N m⁻² h⁻¹ and was found throughout the root zone to a depth of 10 cm. The root zone accounted for as much as 50 to 70% of the annual denitrification. A significant release of organic substrates from the roots seemed to determine the high activities of root zone denitrification in the summer. The denitrification in the surface layer and in the root zone formed two distinct activity zones in the summer, when the root zone also contained nitrification activity, as indicated from the accumulations of NO₃⁻. Light conditions inhibited denitrification in both the surface layer and the upper part of the root zone, suggesting that a release of O₂ by benthic algae and from the roots of L. uniflora controlled a diel variation of denitrification. In midsummer, the rate of denitrification in both the surface layer and the upper part of the root zone was limited by NO₃⁻. In the growth season, there was evidence for a significant population of denitrifiers closely associated with the root surface.     

Influence of Mn2+ ion coordination on tRNAVal1 macrostructure and determination of some coordination sites of Mn2+ ions in tRNAVal1

Electron paramagnetic resonance spectroscopy has been used to study the coupling of Mn²⁺ ions with the tRNA^Val₁ modified with a spin label at four pseudouridylic residues and with the valyl-tRNA^Val₁ modified with a spin label at the α-amine group of the valyl residue. A sharp increase of spin-label mobility has been found in these samples, due to the conformational transition induced by the first and second Mn²⁺ ions. Analysis of dipole–dipole couplings of spin labels with the coordinated ions revealed a definite order in the occupation of ion coordination sites in the tRNA. For some valyl-tRNA^Val₁ molecules, the second Mn²⁺ ions were shown to coordinate on the α-amine group of the valyl residue at a distance of 15–25 Å from a spin label. As a result of the conformational transition, a coordination site appeared in the tRNA at one of the pseudouridylic residues, its distance from the spin label being less than 10 Å. It has been suggested that the conformational transition induced by ions excluded some bases from the system of hydrogen bonds at the level of the tRNA tertiary structure. As a result, these bases acquired sufficient sterical freedom to participate in the Mn²⁺ ion coordination.     

High density culture of anchorage-dependent animal cells by polyurethane foam packed-bed culture systems

Summary Monkey kidney cells (Vero) and Chinese hamster ovary cells (CHO-K1) attached to the internal surface of polyurethane foam (PUF) and grew to a high cell density (1.1 × 10⁸ cells/cm³ PUF and 4.2 × 10⁷ cells/cm³ PUF, respectively) in a PUF-plates packed-bed culture system. This density of Vero cells was twice that obtained previously with a PUF-particles packed-bed culture system. A maximum cell density of 6.7 × 10⁷ cells/cm³ culture vessel volume was obtained in a PUF-disc packed-bed culture of Vero cells. From the cell density of CHO-K1, growing in a monolayer on the surface of PUF and a petri dish, per bulk volume of PUF, we estimated that a surface area to volume ratio of PUF plates effective for cell growth was about 109 cm²/cm³.Offprint requests to: K. Funatsu     

Evaluation of inert and organic carriers for <Emphasis Type="Italic">Verticillium lecanii</Emphasis> spore production in solid-state fermentation

Growth and sporulation of Verticillium lecanii on inert and organic carriers (sugar-cane bagasse, corncob, rice straw, polyurethane foam and activated carbon) in a solid-state fermentation process was studied. Sugar-cane bagasse and polyurethane foam produced 10¹⁰ spores g⁻¹ dry carrier whereas corncob, rice straw, and activated carbon yielded, respectively 8 × 10⁹, 4 × 10⁹, and 3 × 10⁸ spores g⁻¹. Chitinase activity of the conidia was in the following order: sugar-cane bagasse (3.3 U mg⁻¹) > wheat bran (3.0 U mg⁻¹) > polyurethane foam (2.7 U mg⁻¹). There was no significant difference (2.5–2.7 U mg⁻¹) in the proteinase activity among the conidia from the three cultures. Scanning electron microscopy shows that aerial mycelium freely penetrated into the internal area of polyurethane foam. Sugar-cane bagasse provided enough area for vegetative hyphae to attach. Of the carriers analyzed, polyurethane foams and sugar-cane bagasse were the best carriers for V. lecanii growth and spore production.     

Nonwoven rGO Fiber‐Aramid Separator for High‐Speed Charging and Discharging of Li Metal Anode

Li metal, which has a high theoretical specific capacity and low redox potential, is considered to the most promising anode material for next‐generation Li ion‐based batteries. However, it also exhibits a disadvantageous solid electrolyte interphase (SEI) layer problem that needs to be resolved. Herein, an advanced separator composed of reduced graphene oxide fiber attached to aramid paper (rGOF‐A) is introduced. When rGOF‐A is applied, F^? anions, generated from the decomposition of the LiPF₆ electrolyte during the SEI layer formation process form semi‐ionic C? F bonds along the surface of rGOF. As Li⁺ ions are plated, the “F‐doped” rGO surface induces the formation of LiF, which is known as a component of a chemically stable SEI, therefore it helps the Li metal anode to operate stably at a high current of 20 mA cm^?2 with a high capacity of 20 mAh cm^?2. The proposed rGOF‐A separator successfully achieves a stable SEI layer that could resolve the interfacial issues of the Li metal anode.     

Tissue plasminogen activator (t-PA) production by a high density culture of weakly adherent human embryonic kidney cells using a polyurethane-foam packed-bed culture system

Weakly adherent cells of the 293 line attached well to the internal surface of polyurethane foam (PUF) and grew to the high density of 6.83 × 10⁷ cells/cm³ PUF in stationary culture. The maximum productivity of tissue plasminogen activator (t-PA) was 0.158 IU/10⁶ cells per day. The productivity decreased at the stationary phase of cell growth, so we designed a PUF-plate packed-bed culture system for high density culture and continuous production of t-PA. A maximal cell density of 3.24 × 10⁷ cells/cm³ PUF and a t-PA productivity of 0.326 IU/10⁶cells per day were obtained in 25-day perfusion cultures. Although the cell density decreased to half that in PUF stationary culture, the t-PA productivity increased twofold and was maintained for 25 culture days.