bluuebunny/crossref_metadata_2025 huggingface dataset:
A Study on the Electrical Resistance of the Frog Sartorius Muscle
Authors: Schanne Otto , Kawata Hiroshi , Schäfer Bärbel , Lavallée Marc
Published: May 2004

Four different methods of measuring the resistance of a muscle fiber have been applied to the frog sartorius muscle. The methods, in which the resistance of the microelectrode entered the calculation of the effective resistance of the fiber, resulted in values which were 8 times higher than the resistance values obtained with methods independent of the electrode resistance. A simple cable model of a muscle fiber could not account for the discrepancy in the effective resistance found in these measurements; therefore, an enlarged cable model for a muscle fiber has been proposed, and its biological implications have been discussed. The effective resistance (measured with the two different groups of methods) decreased when the potassium concentration in the bath increased. Using the proposed enlarged cable model for the interpretation of these results, it is shown that not only the membrane resistance but also the myoplasmic resistance decreases with an increasing potassium concentration in the Ringer solution.
Potential, Current, and Ionic Fluxes across the Isolated Retinal Pigment Epithelium and Choroid
Authors: Lasansky Arnaldo , de Fisch Felisa W.
Published: May 2004

A flux chamber was utilized for in vitro studies of a membrane formed by the retinal pigment epithelium and choroid of the eye of the toad (Bufo arenarum and Bufo marinus). A transmembrane potential of 20 to 30 mv was found, the pigment epithelium surface positive with respect to the choroidal surface. Unidirectional fluxes of chloride, sodium, potassium, and calcium were determined in the absence of an electrochemical potential difference. A net transfer of chloride from pigment epithelium to choroid accounted for a major fraction of the mean short-circuit current. A small net flux of sodium from choroid to pigment epithelium was detected in Bufo marinus. In both species of toads, however, about one-third of the mean short-circuit current remained unaccounted for. Manometric determinations of bicarbonate suggested an uptake of this ion at the epithelial surface of the membrane but did not provide evidence of a relationship between this process and the short-circuit current.
Blockage of Sodium Conductance Increase in Lobster Giant Axon by Tarichatoxin (Tetrodotoxin)
Authors: Takata M. , Moore J. W. , Kao C. Y. , Fuhrman F. A.
Published: May 2004

Tarichatoxin, isolated from California newt eggs, has been found to selectively block the increase of sodium conductance associated with excitation in lobster giant axons at nanomolar concentrations. This resulted from a reduction in the amplitude of the conductance increase rather than a change in its temporal characteristics. The normal potassium conductance increase with depolarization is not altered. A high concentration of calcium applied concomitantly with the toxin significantly improves the reversibility of the sodium blocking. This toxin has recently been identified as chemically identical with tetrodotoxin from the puffer fish. Toxins from the two sources are equally effective and are shown to have an action which is distinctly different from that of procaine.
The Arrangement of Information in DNA Molecules
Authors: Thomas Charles A.
Published: May 2004

The anatomy of DNA molecules isolated from mature bacteriophage is reviewed. These molecules are linear, duplex DNA consisting mainly of uninterrupted polynucleotide chains. Certain phage (T5 and PB) contain four specifically located interruptions. While the nucleotide sequence of most of these molecules is unique (T5, T3, T7, λ), some are circular permutations of each other (T2, T4, P22). Partial degradation of these DNA molecules by exonuclease III predisposes some of them to form circles upon annealing, but indicating they are terminally redundant.
Problems in Protein Biosynthesis
Authors: Lengyel Peter
Published: May 2004

Outline of the steps in protein synthesis. Nature of the genetic code. The use of synthetic oligo- and polynucleotides in deciphering the code. Structure of the code: relatedness of synonym codons. The wobble hypothesis. Chain initiation and N-formyl-methionine. Chain termination and nonsense codons. Mistakes in translation: ambiguity in vitro. Suppressor mutations resulting in ambiguity. Limitations in the universality of the code. Attempts to determine the particular codons used by a species. Mechanisms of suppression, caused by (a) abnormal aminoacyl-tRNA, (b) ribosomal malfunction. Effect of streptomycin. The problem of "reading" a nucleic acid template. Different ribosomal mutants and DNA polymerase mutants might cause different mistakes. The possibility of involvement of allosteric proteins in template reading.
Membrane Structure and Function
Authors: Kennedy Eugene P. , Fox C. Fred , Carter James R.
Published: May 2004

An understanding of the biochemical basis of membrane function is an important goal of present day biology. In this paper, a biochemical approach to the problem of the specific transport of sugars across the membrane of Escherichia coli is discussed. A new biochemical model for lactose transport system in this organism is presented, in which a specific membrane protein (M protein) plays the role of the sugar carrier. Experiments which have led to the discovery of such a protein, its specific labeling, and partial purification are briefly reviewed.
Methylation of DNA
Authors: Gold Marvin , Gefter Malcolm , Hausmann Rudolph , Hurwitz Jerard
Published: May 2004

The methylated bases of DNA are formed by the transfer of the methyl group from S-adenosylmethionine to a polynucleotide acceptor. This transfer is catalyzed by highly specific enzymes which recognize a limited number of available sites in the DNA. The mechanism for the recognition is presently unknown. In some instances, there is evidence that other cellular components, such as lipopolysaccharides, can influence the methylation reaction. Certain bacteriophages induce new methylases upon infection of their hosts. Phage T3 is unique in establishing an environment in which methylation of neither the phage nor the host nucleic acid can occur. By superinfecting T3-infected cells with other phages, the latter can be obtained with methyl-deficient DNA. Although a great deal is known about the enzymology of the methylation reaction, and there appears to be a strong correlation between the in vitro and in vivo reactions, studies in which DNA is either supermethylated or totally unmethylated have not yielded any insight as to what the possible function of the methylated bases may be.
Analysis of Depolarizing and Hyperpolarizing Inactivation Responses in Gymnotid Electroplaques
Authors: Bennett Michael V. L. , Grundfest Harry
Published: May 2004

In electroplaques of several gymnotid fishes hyperpolarizing or depolarizing currents can evoke all-or-none responses that are due to increase in membrane resistance as much as 10- to 12-fold. During a response the emf of the membrane shifts little, if at all, when the cell either is at its normal resting potential, or is depolarized by increasing external K, and in the case of depolarizing responses when either Cl or an impermeant anion is present. Thus, the increase in resistance is due mainly, or perhaps entirely, to decrease in K permeability, termed depolarizing or hyperpolarizing K inactivation, respectively. In voltage clamp measurements the current-voltage relation shows a negative resistance region. This characteristic accounts for the all-or-none initiation and termination of the responses demonstrable in current clamp experiments. Depolarizing inactivation is initiated and reversed too rapidly to measure with present techniques in cells in high K. Both time courses are slowed in cells studied in normal Ringer's. Once established, the high resistance state is maintained as long as an outward current is applied. Hyperpolarizing inactivation occurs in normal Ringer's or with moderate excess K. Its onset is more rapid with stronger stimuli. During prolonged currents it is not maintained; i.e., there is a secondary increase in conductance. Hyperpolarizing inactivation responses exhibit a long refractory period, presumably because of persistence of this secondary increase in conductance.
The Passive Permeability of the Red Blood Cell to Cations
Authors: LaCelle Paul L. , Rothstein Aser
Published: May 2004

The efflux of salt from human red blood cells suspended in isotonic sucrose plus low concentrations of salt, was measured under steady-state conditions. The relationship between the efflux and the log of the salt concentration can be fitted by two straight lines with a sharp inflection point, the steeper slope occurring at concentrations below 0.2 mM NaCl. The determining factor in the rate of efflux is the ionic strength rather than the specific monovalent cations or anions and the effects are completely reversible. With an increase in temperature, the effects of reduced ionic strength are more pronounced and the inflection point is shifted toward higher salt concentrations. An increase in pH leads to an increased efflux at a given ionic strength, but the size of the pH effect is small at low ionic strength. At a given pH, the data can be fitted by a simplified form of the Goldman equation suggesting that with reduction in ionic strength, the permeability remains constant until the inflection point is reached. At that ionic strength, a sharp reversible transition to a new permeability state occurs. The permeability increases with an increase in the external but not the internal pH.
Discrimination of Single Transport Systems
Authors: Inui Yukiharu , Christensen Halvor N.
Published: May 2004

Uptake of methionine, α-aminoisobutyric acid, and α-(methyl-amino)-isobutyric acid has been shown to occur by at least two transport systems, one sensitive and the other insensitive to the Na+ concentration. For α-aminoisobutyric acid and its N-methyl derivative, the Na+-insensitive uptake is not concentrative and its rate increases almost linearly with concentration within the range examined. In contrast, the Na+-insensitive uptake of methionine is concentrative and subject to inhibition by such amino acids as phenylalanine, leucine, and valine, although not in a manner to indicate that the uptake is mediated by a single agency. This component is not produced by a residual operation of the Na+-requiring transport system, handicapped by the absence of Na+ or by its having combined with α-aminoisobutyric acid. The increase in the rate of methionine uptake is linear with concentration only above about 16 mM methionine. The Na+-sensitive uptakes of methionine, α-aminoisobutyric, and α-(methylamino)-isobutyric acid appear to occur by the same population of transport-mediating sites. Both Km and Vmax of the Na+-sensitive uptake of these three amino acids change with changes in the concentration of Na+, an effect which is shown to have a theoretical basis. A similarity in the values of Vmax for ten amino acids entering principally by the Na+-sensitive agency indicates that differences in their Km values probably measure differences in their affinities for that transport-mediating system.
Cation Effects on Chloride Fluxes and Accumulation Levels in Barley Roots
Authors: Jackson P. C. , Edwards D. G.
Published: May 2004

Accumulation of Cl- by excised barley roots, as of K+, approaches a maximum level at which the ion influx and efflux rates become equal. The rate of Cl- influx at this equilibrium is close to the initial rate while the efflux rate increases with time from zero to equality with influx. The Cl- fluxes are independent of simultaneous exchange flux of the cations, but depend on the nature and concentration of the salt solutions from which they originate. The Cl- content at equilibrium, however, is largely independent of the external concentrations. The approach to equilibrium reflects the presence of the cation. Cl- flux equilibrium is attained more rapidly in KCl than in CsCl or CaCl2. This is presumably an effect of much slower distribution of Cs+ and Ca++ than of K+ within the roots. Accumulated Cs+ appears to form a barrier to ion movement primarily within the outermost cells, thereby reducing influx and ultimately efflux rates of both Cl- and cations. Slow internal mixing and considerable self-exchange of the incoming ions suggest internal transport over a series of steps which can become rate-limiting to the accumulation of ions in roots.
Sodium Extrusion by Internally Dialyzed Squid Axons
Authors: Brinley F. J. , Mullins L. J.
Published: May 2004

A method has been developed which allows a length of electrically excitable squid axon to be internally dialyzed against a continuously flowing solution of defined composition. Tests showed that diffusional exchange of small molecules in the axoplasm surrounding the dialysis tube occurred with a half-time of 2–5 min, and that protein does not cross the wall of the dialysis tube. The composition of the dialysis medium was (mM): K isethionate 151, K aspartate 151, taurine 275, MgCI2 4–10, NaCl 80, KCN 2, EDTA 0.1, ATP 5–10, and phosphoarginine 0–10. The following measurements were made: resting Na influx 57 pmole/cm2sec (n = 8); resting potassium efflux 59 pmole/ cm2sec (n = 4); stimulated Na efflux 3.1 pmole/cm2imp (n = 9); stimulated K efflux 2.9 pmole/cm2imp (n = 3); resting Na efflux 48 pmole/cm2sec (n = 18); Q10 Na efflux 2.2 (n = 5). Removal of ATP and phosphoarginine from the dialysis medium (n = 4) or external application of strophanthidin (n = 1) reversibly reduced Na efflux to 10–13 pmole/cm2sec. A general conclusion from the study is that dialyzed squid axons have relatively normal passive permeability properties and that a substantial fraction of the Na efflux is under metabolic control although the Na extrusion mechanism may not be working perfectly.
Effect of Inhibitors on Alanine Transport in Isolated Rabbit Ileum
Authors: Chez Ronald A. , Palmer Richard R. , Schultz Stanley G. , Curran Peter F.
Published: May 2004

The effects of metabolic inhibitors and ouabain on alanine transport across rabbit ileum, in vitro, have been investigated. Net transport of alanine and Na across short-circuited segments of ileum is virtually abolished by cyanide, 2,4-dinitrophenol, iodoacetate, and ouabain. However, these inhibitors do not markedly depress alanine influx from the mucosal solution, across the brush border, into the intestinal epithelium, and they do not significantly affect the Na dependence of this entry process. The results of this investigation indicate that: (a) the Na dependence of alanine influx does not reflect a mechanism in which the sole function of Na is to link metabolic energy directly to the influx process; and (b) the inhibition of net alanine transport across intestine is, in part, the result of an increased rate coefficient for alanine efflux out of the cell across the brush border. Although these findings do not exclude a direct link between metabolic energy and alanine efflux, the increased efflux may be the result of the increased intracellular Na concentration in the presence of these inhibitors. The results of these studies are qualitatively consistent with a model for alanine transport across the brush border which does not include a direct link to metabolic energy.
On the Ability of Isolated Frog Skin to Manufacture Ringer's Fluid
Authors: Steinbach H. Burr
Published: May 2004

Fresh, empty, isolated sacs of leg skin from R. pipiens manufacture a salt solution reasonably balanced with respect to ions and approximating a somewhat dilute Ringer's fluid. Concentrations of ions bear little relationship to concentrations in the external medium even down to 12 mM. An effective regulatory mechanism is indicated whereby the amount of salt transported is adjusted to the amount of water or vice versa, the rate of movement of either salt or water being largely independent of osmotic or ionic gradients (outer fluid to manufactured inner fluid). Concentrations of the inner fluids appear to be regulated to conform to a fairly constant concentration within the skin. Some evidence is presented that a major factor in regulating concentration of the inner fluid is triggered by an initial dilution of the inner fluid, followed by stimulation of salt uptake.
The Effect of Valinomycin on Potassium and Sodium Permeability of HK and LK Sheep Red Cells
Authors: Tosteson Daniel C. , Cook P. , Andreoli Thomas , Tieffenberg M.
Published: May 2004

A cyclic depsipeptide antibiotic, valinomycin, was found to produce increased selective permeability of the plasma membranes of HK and LK sheep red blood cells to potassium but not to sodium ions. The compound had relatively little effect on the active extrusion of sodium from HK sheep red blood cells or on the Na + K-stimulated ATPase activity of membranes derived from these cells. It is proposed that the selective cation permeability produced by this compound depends primarily on steric factors, particularly the relationship between the diameter of the ring and the effective diameter of the ion. The significance of these results for the problem of the mechanism of ionic selectivity in natural membranes is discussed.
The Effect of Valinomycin on the Ionic Permeability of Thin Lipid Membranes
Authors: Andreoli Thomas E. , Tieffenberg M. , Tosteson Daniel C.
Published: May 2004

Optically black membranes prepared from sheep red cell lipids have a high electrical resistance (1–3 x 108 ohm-cm2). The ionic transference numbers (Ti) for cations (Na+ or K+) are equal to each other but at least four to five times greater than for Cl-. The cyclic depsipeptide valinomycin produces a striking decrease in the membrane resistance when K+, but not when Na+ is in the solutions bathing the membrane. The ratio TNa/TK, estimated from membrane voltages in the presence of ionic concentration gradients, approaches zero. The order of membrane monovalent cation selectivity, in the presence of valinomycin, is H+ > Rb+ > K+ > Cs+ > Na+. Addition of the antibiotic to one side of a membrane which separates identical solutions of NaCl produces a substantial (up to 80 mV) membrane voltage (side opposite valinomycin negative). These data are consistent with the hypothesis that valinomycin can interact with appropriately sized cations (hydrated diameter ??? 6 A) to increase their membrane permeability, perhaps by forming hydrogen bonds between the solvation shell of the cations and carbonyl oxygens in the valinomycin molecule which are directed toward the aperture of the ring.
Nonsolvent Water in Human Erythrocytes and Hemoglobin Solutions
Authors: Bobo C. M. Gary
Published: May 2004

Distribution ratios of water-soluble nonelectrolytes have been measured for two systems, human red bood cells/isotonic saline and hemoglobin solutions/water. The results show that for these solutes there is a significant amount of nonsolvent water associated with Hb. However, the amount of this nonsolvent water depends markedly on the temperature, as well as on the size, steric configuration, and functional groups of the probe solutes. The significance of the data is discussed.
Fluorometric Measurement of Pyridine Nucleotide Reduction in the Giant Axon of the Squid
Authors: Doane Marshall G.
Published: May 2004

By monitoring the fluorescence of the isolated giant axon of the squid Loligo pealei, it was possible to follow changes in its oxidation-reduction state as caused by the action of anoxia, cyanide, Amytal, and azide. The response to oxygen depletion was very rapid, the NAD within the axon being 90% reduced within 1–2 min. Cyanide and Amytal gave essentially similar results, although somewhat longer periods of time elapsed during their onset and washout periods. The extent of NAD reduction was essentially the same under conditions of anoxia and treatment with cyanide and Amytal. Azide was less effective in this respect, and at comparatively high levels of concentration (25–50 mM) gave values of 40% or less of the reduction observed with the other inhibitors. The application of ouabain and strophanthidin gave no observable NAD reduction. Variations in the time required to consume given quantities of dissolved oxygen before and after stimulation indicated an increase of 10–20% in oxygen uptake rate associated with activity, although this figure appeared to be a function of the surface-to-volume ratio of the axon. A biochemical analysis of axoplasm for oxidized and reduced pyridine nucleotide was made. Fluorometric examination of centrifuged axoplasm indicated that the NAD-NADH was largely confined to the mitochondria of the axon.
Ca Fluxes in Single Twitch Muscle Fibers
Authors: Curtis B. A.
Published: May 2004

Ca influx and efflux in single twitch muscle fibers were determined by the movement of 45Ca. The isotope was assayed by counting the center 1 cm of a fiber while it was in nonradioactive Rnger's solution. The average resting influx in 1.0 mM Ca Ringer's was 0.26 pM Ca/cm2. sec for 5 to 20 min influx periods. The average additional influx upon stimulation in 1.0 mM Ca was 0.73 pM Ca/cm2. twitch. The efflux after both resting and stimulated 45Ca influx can be described by a single exponential curve with an average time constant of 125 min. This relationship is an indication of Ca exchange with a single intracellular compartment. This compartment contains an estimated 47% of the total muscle Ca at 1.0 mM Ca. When the Ca in the Ringer was reduced to 0.5 mM Ca, both the resting and stimulated Ca fluxes decreased. When Ca was raised to 1.8 mM, the stimulated influxes increased but the resting influx did not.
Voltage Clamp Studies on the Effect of Internal Cesium Ion on Sodium and Potassium Currents in the Squid Giant Axon
Authors: Adelman William J. , Senft Joseph P.
Published: May 2004

Isolated and cleaned giant axons of Loligo pealii were internally perfused with solutions containing cesium sulfate and potassium fluoride. Membrane currents obtained as a function of clamped membrane potentials indicated a severe depression of the delayed outward current component normally attributed to potassium ion movement. Steady-state currents showed a negative slope in the potential range from -45 to -5 mv which corresponded to the negative slope for the peak sodium current relation vs. membrane potential which suggested long duration sodium currents. Using sodium-free sea water externally, sodium currents were separated from total currents and these persisted for longer times than normal. This result suggested that internal cesium, ion delays the sodium conductance turnoff. The separated nonsodium currents showed an abnormal rectification as compared with those predicted by the independence principle, such that while potassium permeability appeared normal at the resting potential, its value decreased progressively with increasing depolarization.
Stimulation of the Salt Receptor of the Blowfly
Authors: Gillary Howard L.
Published: May 2004

Application of NaCl solutions to the tip of a labellar sensillum of the blowfly elicited a repetitive neural response from the salt receptor. The response was examined with respect to reproducibility and adaptation. A threshold was observed for tests with dilute solutions. Above this, the response increased linearly with the logarithm of the molarity. The response was not significantly affected by the pH of stimulating solutions, buffered or not, between 3 and 10. Beyond this range, it was reversibly inhibited until, at greater extremes of pH, atypical stimulation independent of the presence of salt was seen. Receptor sensitivity increased with fly age. The results presented here may be due to effects at sites in the sensillum other than the receptor membrane.
Increased Mutation Rate of E. coli K12λ Cultures Maintained in Continuous Logarithmic Growth
Authors: Northrop John H.
Published: May 2004

Continuous logarithmic growth of E. coli K12λ in an automatic culture cell resulted in marked increases in the proportion of several mutants. The P1 phage-resistant cells increased 10 to 3000 times, the T2 phage-resistant cells 1 to 1000 times, the neomycin-resistant cells 1 to 10 times, and the virus-producing cells 30 to 70 times. No change occurred in the penicillin-resistant cells. Calculation of the growth curves and direct determination of the mutation rates by the null fraction method showed that the increases in the proportion of mutants were due to increases in the mutation rates.
Anion Permeability of the Olfactory Receptive Membrane
Authors: Takagi S. F. , Wyse G. A. , Yajima Toshi
Published: May 2004

The ionic mechanism of the electropositive olfactory receptor potential was studied in the bullfrog and the swamp frog. The positive receptor potential strikingly decreased in amplitude in chloride-free solution. When the olfactory epithelium was immersed in high-KCl-Ringer's solution and then in Cl-free, high-K solution, the polarity of the positive potential could be reversed. This is supposed to be due to the exit of the increased internal chloride ion. From the above two experiments it is concluded that the positive olfactory receptor potential depends primarily upon the influx of the chloride ion through the olfactory receptive membrane. Some contribution by potassium and possibly other ions may occur. The ability of other anions to substitute for chloride was examined. It was found that only Br-, F-, and HCO2- could penetrate the olfactory receptive membrane. The sieve hypothesis in the inhibitory post-synaptic membrane (Coombs, Eccles, and Fatt, 1955) is not applicable to the olfactory receptive membrane on the basis of the size of hydrated ions, but it may be applicable on the basis of the sizes of naked ions.
Mach Band Type Lateral Inhibition in Different Sense Organs
Authors: von Békésy Georg
Published: May 2004

Experiments were done on the skin with shearing forces, vibrations, and heat stimuli and on the tongue with taste stimuli to show that the well known Mach bands are not exclusively a visual phenomenon. On the contrary, it is not difficult to produce areas of a decreased sensation magnitude corresponding to the dark Mach bands in vision. It is shown on a geometrical model of nervous interaction that the appearance of Mach bands for certain patterns of stimulus distribution is correlated with nervous inhibition surrounding the area of sensation. This corroborates the earlier finding that surrounding every area transmitting sensation there is an area simultaneously transmitting inhibition.
Enhanced Permeability to Sugar Associated with Muscle Contraction
Authors: Holloszy John O. , Narahara H. T.
Published: May 2004

When contractures were induced in isolated frog sartorius muscles with 4 mM caffeine, there was an increase in permeability of the muscle cells to 3-methylglucose. This observation suggests that the changes in permeability to sugar that are known to occur in electrically stimulated muscles may not be intimately related to the depolarization phase of the tissue response. Contractures that were elicited by exposing the muscles to a high concentration of K+ were also associated with an increased permeability to sugar. As the concentration of 45Ca in the medium was raised, more 45Ca entered the muscles during potassium contractures, and the contractures lasted longer, in agreement with the observations of other investigators. There was also a greater change in permeability to sugar when potassium contractures were elicited in the presence of higher concentrations of Ca++. The possibility that the enhanced permeability to sugar may be related to changes in the intracellular concentration of Ca++ is discussed.
The Electrical and Mechanical Activity of the Esophageal Cell of Ascaris lumbricoides
Authors: del Castillo J. , Morales T.
Published: May 2004

The esophagus of Ascaris is a syncytial muscle organ of tubular shape in which the myofibrils are arranged radially between the lumen and the external surface. A resting potential of almost 40 mv (cytoplasm negative) is maintained by the extracellular organic anions (volatile fatty acids) found in the perienteric fluid. Replacement of these anions by Cl- ions results in a large depolarization. The resting potential is also decreased when the external pH is lowered. The leading phase of the action potential with a positive overshoot of about 18 mv elicits contraction of the myofibrils, development of negative pressure within the lumen, and suction of liquid and food particles. The mechanical energy stored in the elastic components of the cell is released when the myofibrils relax, thus injecting the contents of the lumen into the intestine. A fast and synchronous relaxation is elicited by a regenerative membrane polarization, a negative spike with a peak value of up to 108 mv produced by an increase in the permeability of the membrane to K+ ions. Cells completely depolarized in "chloride" saline are still able to generate such large potassium spikes.
The Circadian Rhythm in Photosynthesis in Acetabularia in the Presence of Actinomycin D, Puromycin, and Chloramphenicol
Authors: Sweeney Beatrice M. , Tuffli Charles F. , Rubin Richard H.
Published: May 2004

Anucleate Acetabularia crenulata shows a circadian rhythm in photosynthesis. In this study, an oxygen electrode was employed to measure this photosynthetic rhythm in the presence and absence of the inhibitors, actinomycin D, chloramphenicol, and puromycin. High concentrations of the inhibitors were used: actinomycin D, 20–40 µg ml-1 puromycin, 30 and 100 µg ml-1; and chloramphenicol, 250 µg ml-1. The effectiveness of these inhibitors on protein synthesis was also measured under the same conditions used for the determination of rhythmicity. In spite of large effects of all three inhibitors on the incorporation of 14C leucine, no effect on the period or the phase of the photosynthetic rhythm was observed. The higher concentration of puromycin and chloramphenicol produced toxic effects which were expressed as a reduction in the amount of photosynthesis, but rhythmicity was still apparent. After 3 or 4 days' exposure to actinomycin, Acetabularia became resistant to its effect. Recovery was also observed in the ability to incorporate leucine. The implications of these results for theories of the basic oscillator responsible for circadian rhythmicity are discussed.
The Extracellular Space of Voluntary Muscle Tissues
Authors: Ling Gilbert N. , Kromash Marsha H.
Published: May 2004

The volume occupied by the extracellular space has been investigated in six types of voluntary muscles: sartorius (frog), semitendinosus (frog), tibialis anticus longus (frog), iliofibularis (frog), rectus abdominis (frog), and diaphragm (rat). With the aid of four types of probe material, three of which are conventionally employed (inulin, sorbitol, sucrose) and one of which is newly introduced (poly-L-glutamate), and a different experimental method, we have demonstrated that the "true" extracellular space of frog sartorius, semitendinosus, tibialis anticus longus, and iliofibularis muscle and of rat diaphragm muscle is equal to, or probably less than, 8–9% (v/w) of the tissue. The frog rectus muscle shows a somewhat higher ceiling value of 14%.
Permeability Changes Associated with the Action Potential in Procaine-Treated Crayfish Abdominal Muscle Fibers
Authors: Takeda Kimihisa
Published: May 2004

Permeability changes associated with prolonged action potentials have been analyzed in procaine-treated crayfish abdominal muscle fibers. The effect of external Ca indicates that the increase in membrane conductance observed during the rising phase of the action potential is primarily due to a permeability increase for Ca. A remnant of the permeability increase may cause the succeeding plateau as shown by its high conductance and by the effect of low Mn. A delayed increase in conductance precedes the termination of the plateau phase. This is due to a delayed increase in permeability, probably for K, that is observed when depolarizing electrogenesis is eliminated. High external Ca reduces the action potential duration, the falling phase starting at a higher depolarization. These changes may be related to an earlier onset of the delayed increase in permeability, induced by a larger inside positivity in the presence of higher Ca. No "anomalous rectification" is seen in early or late I-V curves for small depolarizations. Ba may replace Ca in its role in depolarizing electrogenesis, and the first action potential induced in Ba saline has a large overshoot and a long duration. In higher Ba salines, action potentials are greatly prolonged. Long term soaking in Rb-containing or K-free saline also augments and prolongs the action potential. These changes are assumed to be related to depression of the K permeability of the membrane.
The Influence of Ionic Strength on Potassium Contractures and Calcium Movements in Frog Muscle
Authors: Lorković Hrvoje
Published: May 2004

Frog toe muscles were bathed in isotonic, sodium-free Tris chloride, methanesulfonate, or sulfate solutions containing sucrose or mannitol and varying in ionic strength from 0.006 to 0.291. By decreasing the ionic strength the curve relating the peak tension of the K contractures to the log [K] was reversibly shifted to lower [K]. Increasing the [Ca] from 1 to 4 mM almost abolished this effect. The resting uptake of 45Ca was increased more than two times by decreasing the ionic strength from 0.125 to 0.039. It was not increased significantly by raising [Ca] from 1 to 4 mM at low or normal ionic strength. The additional uptake of 45Ca during contractures provoked by 120 mM K was not significantly different at the two levels of ionic strength. The rate of emergence of 45Ca from muscles loaded with 45Ca at reduced ionic strength, was decreased. The effects of low ionic strength are discussed in terms of changes in the potential difference across a membrane with fixed negative charges on the outer surface.
Cation Metabolism in Relation to Cell Size in Synchronously Grown Tissue Culture Cell
Authors: Jung Chan , Rothstein Aser
Published: May 2004

In randomly grown tissue culture cells (mouse leukemic lymphoblast, L5178Y) the number, volume, and Na+ and K+ content increase as an exponential function with a doubling time of 11.3 hr. In synchronously grown cells the volume increase of the population and of single cells follows the same exponential function as in randomly grown cells. In contrast, the cation content fluctuates during a single cell cycle. About 1½ hr after the cell division burst (at the beginning of the S period), a net loss of K+ occurs for a period of about 1 hr amounting to about 20% of the total K. Over the next 5 to 6 hr, the deficit in K+ is eliminated. The Na+ content shows a double fluctuation. It falls during the cell division burst, rises when the K+ content decreases, falls again when K+ content rises, and then increases again before the next cell division burst. The net fluxes of both Na+ and K+ are very small compared to the unidirectional fluxes (less than 5%), thus small changes in the balance of influx and efflux account for the changes in cation content during the growth cycle. Both unidirectional fluxes increase dramatically (by a factor of two) about 2 hr after the cell division burst, and then remain constant until after the next cell division. The pattern of electrolyte regulation during cell division does not follow a simple function such as cell number, cell surface, or cell volume, but must be related to specific internal events in the cell.
Hyperosmolarity and the Net Transport of Nonelectrolytes in Frog Skin
Authors: Franz T. J. , Van Bruggen J. T.
Published: May 2004

The permeability of frog skin to a series of nonelectrolytes (thiourea, urea, mannitol, and sucrose) under the influence of 2.5 times normal osmolarity in the outer bathing solution has been investigated. Although the flux of the tracer nonelectrolytes across the skin in either direction is greatly increased by hyperosmolarity, the influx is found to be increased to a significantly greater extent than the outflux. Flux ratios as high as 3:1 can be observed. The net inward movement of the nonelectrolyte proceeds in spite of a sizeable bulk flow of water in the opposite direction. Possible driving forces for this phenomenon are discussed.
Validity of the Force-Velocity Relation for Muscle Contraction in the Length Region, l ≤ l0
Authors: Matsumoto Yorimi
Published: May 2004

Considerable attention has been directed to the characteristic force-velocity relation discovered by A. V. Hill in the study of muscle kinematics. Models of contractile process were tested on the basis of their compatibility with the Hill equation. However, almost all the isotonic data have been restricted to one length, l0, the maximum length with almost no resting tension; the velocities measured are those initial values when the load begins to move. The force-velocity curve extrapolates to zero velocity for isometric tension, but only for the tension at that one length. Very few efforts have been made to study the profiles of the curves throughout the range of lengths over which shortening takes place. In examining the length region, l ≤ l0, for an isotonically contracting muscle, not only is the force-velocity relation valid for the initial reference length, l0, but also for any other length. The analysis in this report indicates that the constants a/P0 and b/l0 remain fixed throughout the length change of afterloaded isotonic shortening in the Rana pipiens sartorius muscles.
Theoretical Series Elastic Element Length in Rana pipiens Sartorius Muscles
Authors: Matsumoto Yorimi
Published: May 2004

Assuming a two component system for the muscle, a series elastic element and a contractile component, the analyses of the isotonic and isometric data points were related to obtain the series elastic stiffness, dP/dls, from the relation,

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From the isometric data, dP/dt was obtained and shortening velocity, v, was a result of the isotonic experiments. Substituting (P0 - P)/T for dP/dt and (P0 - P)/(P + a) times b for v, dP/dls = (P + a) /bT, where P < P0, and a, b are constants for any lengths l ≤ l0 (Matsumoto, 1965). If the isometric tension and the shortening velocity are recorded for a given muscle length, l0, although the series elastic, ls, and the contractile component, lc, are changing, the total muscle length, l0 remains fixed and therefore the time constant, T. Integrating,

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the stress-strain relation for the series elastic element,

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is obtained; lsc0 - ls + lc0where lco equals the contractile component length for a muscle exerting a tension of P0. For a given P/P0, ls is uniquely determined and must be the same whether on the isotonic or isometric length-tension-time curve. In fact, a locus on one surface curve can be associated with the corresponding locus on the other.
Acetylcholine and Calcium on Membrane Permeability and Contraction of Intestinal Smooth Muscle
Authors: Hurwitz Leon , Von Hagen Stanley , Joiner Paul D.
Published: May 2004

Acetylcholine elicited a sustained contraction and an increase in potassium efflux in longitudinal muscle isolated from the guinea pig ileum. Stepwise increases in the calcium concentration of the bathing medium, from 0.06 to 36 mM generally reduced the increase in potassium efflux, but had a complex effect on the mechanical response. Contractions produced by high levels of acetylcholine became progressively larger or remained at a high magnitude as the calcium concentration was increased. Contractions produced by low levels of acetylcholine also improved initially, but were depressed again by the highest concentration of calcium introduced. Ethanol, in the appropriate concentration, inhibited completely the acetylcholine-induced contraction without reducing the increase in potassium efflux. Calcium reversed this effect. Both extracellular calcium and ethanol depressed the large, transient increase in muscle tone developed by fibers that were preincubated in a high calcium medium and then exposed to a calcium-free medium. These findings suggested that extracellular calcium ions react with two different sites in the membrane, a stabilizing site and a storage site. A muscle contraction is activated by calcium ions which diffuse from the storage site to the myoplasm. Calcium ions reacting with the stabilizing site impede this diffusion process. Part of the stimulatory effect of acetylcholine is derived from its capacity to counteract the action of calcium at the stabilizing site.
Sodium Exchange in Dog Ventricular Muscle
Authors: Langer G. A.
Published: May 2004

Sodium exchange was studied in the arterially perfused papillary muscle of the dog. Three kinetically defined phases accounted for all the myocardial sodium: phase 0 (vascular)-λo (exchange constant) = 3.6 min-1 phase 1 (interstitial)-λ1 = 0.62 min-1; phase 2 (intracellular)-λ2 < 0.020 min-1 in quiescent muscles. The phase 2 exchange rate was proportional to frequency of contraction and increased by approximately 0.004 min-1 for each 1 beat/min increment in rate in muscles demonstrating stable function. A sudden increase in frequency of contraction was followed by a marked increase in phase 2 sodium exchange if muscle function did not deteriorate. This increased exchange required 14 min to achieve a steady state. During this time active tension increased (positive staircase) and then declined to become stable as the sodium exchange stabilized. In muscles in which increased frequency of contraction produced a progressive decrease in active tension and contracture, sodium exchange failed to increase. The characteristics of sodium exchange are compared to those previously defined for calcium and potassium in the perfused dog papillary muscle. It is proposed that alteration in sodium exchange is a primary determinant of calcium and potassium movements and thereby plays a significant role in the control of myocardial contractility.
Binding of Ca Ions by Paramecium caudatum
Authors: Naitoh Yutaka , Yasumasu Ikuo
Published: May 2004

Binding of 45Ca by live Paramecium caudatum was determined under various external ionic conditions. It was found that calcium uptake was separable into at least two components, a rapid and a slow one. The rapid component was influenced by the presence of certain other ions in a manner which agrees with the law of mass action. It appears that an ion exchange system may be involved in a binding equilibrium established between Paramecium, Ca++, and certain other ions. K+, Rb+, and Ba++ in the equilibrium medium are among those ions which inhibit calcium uptake. It is proposed that liberation of Ca++ from binding sites on Paramecium by an exchange reaction with competing ions is the first step in the mechanism of ciliary reversal in the response to external application of these ions.
Effects of ATP on the Interaction of Ca++, Mg++, and K+ with Fragmented Sarcoplasmic Reticulum Isolated from Rabbit Skeletal Muscle
Authors: Carvalho Arselio P. , Leo Barbara
Published: May 2004

Fragmented sarcoplasmic reticulum isolated from skeletal muscle of the rabbit has a cation-binding capacity of about 350 µeq/g of protein at neutral pH. The same binding sites bind Ca, Mg, K, and H ions and, consequently, the selective binding of Ca induced by ATP releases an amount of the other cations equivalent to the Ca taken up. At pH values below 6.2, an increasing number of binding sites are associated with H+, and ATP induces exchange of Ca mostly for H+. At pH values above 6.2, the binding sites exist in the form of Mg and K, and Ca is bound in exchange for these cations. The total bound Ca + Mg + K, expressed in microequivalents of cations bound per gram of protein, is approximately constant at various pCa values, which indicates a stoichiometric exchange of Ca for the other cations. To accomplish the same degree of exchange of Ca for other cations bound, in the absence of ATP, concentrations of free Ca++ of about 1000-fold higher than those needed in the presence of ATP are required in the medium. We cannot distinguish between a mechanism whereby Ca actively transported into a compartment of the microsomal vesicles containing also the binding sites is bound passively to these sites in exchange for Mg, K, and H and another in which ATP selectively increases the affinity of surface-binding sites for Ca. Irrespective of the mechanism of accumulation, the Ca retained does not contribute to the activity of the cation in the membrane fraction. Caffeine (10 mM) has no effect on the binding of Ca, but releases a more labile fraction of Ca, which presumably accumulates in excess of the bound Ca. Procaine (5 mM) antagonizes the effect of caffeine. Acetylcholine and epinephrine have no effect on the binding of Ca.
The Effect of the Unstirred Layer on Human Red Cell Water Permeability
Authors: Sha'afi R. I. , Rich G. T. , Sidel Victor W. , Bossert W. , Solomon A. K.
Published: May 2004

A study has been made of water entry into human red blood cells under an osmotic pressure gradient. The measurements were made using a rapid reaction stop flow apparatus, whose construction, calibration, and performance are described in detail. Red cell volume changes were determined from 90° scattered light. The permeability coefficient for water entry under a relative isosmolar concentration of 1 to 1.5 was found to be 0.22 ± 0.01 cm4/sec osmol, which agrees well with our previously published value. The experiments were also designed to measure the thickness of the unstirred layer around the6 red cells. This was found to be 5.5 ± 0.8 µ under the present experimental conditions. It is concluded that our previously measured permeability coefficient for water entrance under a diffusion gradient does not require correction on account of the unstirred layer.
Basis of Tetrodotoxin's Selectivity in Blockage of Squid Axons
Authors: Moore John W. , Blaustein Mordecai P. , Anderson Nels C. , Narahashi Toshio
Published: May 2004

The blockage of nerve activity by tetrodotoxin is unusually potent and specific. Our experiments were designed to distinguish whether its specificity of action was based on the identification of ions, the direction of cation flow, or differences in the early transient and late steady conductance pathways. Alkali cations were substituted for sodium in the sea water, bathing an "artificial node" in a voltage-clamped squid axon. When tetrodotoxin was added to the artificial sea waters at a concentration of 100 to 150 mM, it was found to always block the flow of cations through the early transient channel, both inward and outward, but it never blocked the flow of ions using the late steady pathway. We conclude that the selectivity of tetrodotoxin is based on some difference in these two channels.
Comparison of Tetrodotoxin and Procaine in Internally Perfused Squid Giant Axons
Authors: Narahashi Toshio , Anderson Nels C. , Moore John W.
Published: May 2004

Squid giant axons were internally perfused with tetrodotoxin and procaine, and excitability and electrical properties were studied by means of current-clamp and sucrose-gap voltage-clamp methods. Internally perfused tetrodotoxin was virtually without effect on the resting potential, the action potential, the early transient membrane ionic current, and the late steady-state membrane ionic current even at very high concentrations (1,000–10,000 nM) for a long period of time (up to 36 min). Externally applied tetrodotoxin at a concentration of 100 nM blocked the action potential and the early transient current in 2–3 min. Internally perfused procaine at concentrations of 1–10 mM reversibly depressed or blocked the action potential with an accompanying hyperpolarization of 2–4 mv, and inhibited both the early transient and late steady-state currents to the same extent. The time to peak early transient current was increased. The present results and the insolubility of tetrodotoxin in lipids have led to the conclusion that the gate controlling the flow of sodium ions through channels is located on the outer surface of the nerve membrane.