Calcium exchange and contraction strength of guinea pig atrium in normal and hypertonic media

A Krebs-Henseleit (KH) medium made hypertonic by adding nonpermeant molecules substantially increased the isometric peak tension at steady-state contractions below 3 per sec in guinea pig atrium at 27°C. Action potential durations were decreased. KH plus 100 mM raffinose or sucrose resulted in similar and nearly maximal changes which were essentially reversible upon return to normal KH. When one active contracting atrium was used to passively stretch a second atrium, the difference in Ca ion exchange (1 min exchange with the extracellular space) between active and stretched atria significantly increased at 1 per sec and at 2 per sec in going from normal to 100 mM hypertonic KH. The calculated mean Ca ion cellular exchange per beat per 100 g of cells (a) doubled in changing from normal to 100 mM hypertonic KH, and (b) decreased slightly in changing from contractions of 1 per sec to 2 per sec in normal KH. These data are consistent with the hypothesis (a) that Ca ion entry per beat from the extracellular space is proportional to membrane depolarized time with a constant medium and a steady-state condition, and the hypothesis (b) that 100 mM hypertonicity doubles the Ca ion entry rate during depolarization. These data enable rejection of the hypothesis that the peak tension is proportional to the Ca ion entry per beat from the extracellular space under steady-state conditions, and suggest that any additional Ca ion involved in the larger contractions at higher frequencies comes from an increase in Ca ion available from intracellular stores.     

Electroretinogram Characteristics and the Spectral Mechanisms of the Median Ocellus and the Lateral Eye in Limulus polyphemus

Electrical responses (ERG) to light flashes of various wavelengths and energies were obtained from the dorsal median ocellus and lateral compound eye of Limulus under dark and chromatic light adaptation. Spectral mechanisms were studied by analyzing (a) response waveforms, e.g. response area, rise, and fall times as functions of amplitude, (b) slopes of amplitude-energy functions, and (c) spectral sensitivity functions obtained by the criterion amplitude method. The data for a single spectral mechanism in the lateral eye are (a) response waveforms independent of wavelength, (b) same slope for response-energy functions at all wavelengths, (c) a spectral sensitivity function with a single maximum near 520 mµ, and (d) spectral sensitivity invariance in chromatic adaptation experiments. The data for two spectral mechanisms in the median ocellus are (a) two waveform characteristics depending on wavelength, (b) slopes of response-energy functions steeper for short than for long wavelengths, (c) two spectral sensitivity peaks (360 and 530–535 mµ) when dark-adapted, and (d) selective depression of either spectral sensitivity peak by appropriate chromatic adaptation. The ocellus is 200–320 times more sensitive to UV than to visible light. Both UV and green spectral sensitivity curves agree with Dartnall's nomogram. The hypothesis is favored that the ocellus contains two visual pigments each in a different type of receptor, rather than (a) various absorption bands of a single visual pigment, (b) single visual pigment and a chromatic mask, or (c) fluorescence. With long duration light stimuli a steady-state level followed the transient peak in the ERG from both types of eyes.     

Calcium Flux in the Mammalian Ventricular Myocardium

The exchange of Ca⁴⁵ was studied in dog myocardium by means of a newly developed perfusion technique whereby an excised papillary muscle was perfused through its own artery. This makes possible the sequential and simultaneous correlation of ionic flux with ventricular myocardial function with each muscle serving as its own control. Calcium exchange has the following characteristics: (a) The major component of calcium flux is independent of the frequency of contraction. It demonstrates a rapidly equilibrating phase (half-time, 4 to 6 minutes) and a more slowly equilibrating phase with a progressively decreasing rate constant. The flux characteristics of the more rapidly equilibrating compartment are determined by a factor or factors, in addition to simple diffusion, which increase the time required for this compartment to achieve a steady-state with respect to the vascular compartment. (b) A lesser component of exchange is stimulus-rate dependent and is characterized by an alteration in the rate of calcium turnover such that the altered influx: efflux ratio requires 20 to 25 minutes to achieve equilibrium. After this time, despite the higher stimulus rate, there is no evidence of change in total tissue calcium. (c) The initial rate of the transient response is approximately proportional to the change in stimulus rate.     

ADENINE NUCLEOTIDE-INDUCED CONTRACTION OF THE INNER MITOCHONDRIAL MEMBRANE : II. Effect of Bongkrekic Acid

In bovine heart mitochondria bongkrekic acid at concentrations as low as about 4 nmol/mg protein (a) completely inhibits phosphorylation of exogenous adenosine diphosphate (ADP) and dephosphorylation of exogenous adenosine triphosphate (ATP), (b) completely reverses atractyloside inhibition of inner membrane contraction induced by exogenous adenine nucleotides, and (c) decreases the amount of adenine nucleotide required to elicit maximal exogenous adenine nucleotide-induced inner membrane contraction to a level which appears to correspond closely with the concentration of contractile, exogenous adenine nucleotide binding sites Bongkrekic acid at concentrations greater than 4 nmol/mg protein induces inner membrane contraction which seems to depend on the presence of endogenous ADP and/or ATP. The findings appear to be consistent with the interpretations (a) that the inner mitochondrial membrane contains two types of contractile, adenine nucleotide binding sites, (b) that the two sites differ markedly with regard to adenine nucleotide affinity, (c) that the high affinity site is identical with the adenine nucleotide exchange carrier, (d) that the low affinity site is accessible exclusively to endogenous adenine nucleotides and is largely unoccupied in the absence of bongkrekic acid, and (e) that bongkrekic acid increases the affinity of both sites in proportion to the amount of the antibiotic bound to the inner membrane.     

Inhibition of Impulse Activity in a Sensory Neuron by an Electrogenic Pump

The crayfish tonic stretch receptor neuron manifests three phenomena: (a) Impulse frequency in response to a depolarizing current decays exponentially to half the initial rate with a time constant of about 4 sec. (b) One or more extra impulses superimposed on steady activity result in a lengthening of the interspike interval immediately following the last extra impulse which is proportional to the number of extra impulses. However, above a "threshold' number of impulses the proportionality constant becomes abruptly larger. (c) Following trains of impulses, the resting potential of the cell is hyperpolarized by an amount proportional to impulse number. Such posttetanic hyperpolarization (PTH) decays approximately exponentially with a time constant of 11 sec, but this varies with membrane potential. These effects are attributed to the incremental increase of an inhibitory (hyperpolarizing) current with a long (relative to interspike interval) decay constant. We suggest that this inhibitory current is the result of increased electrogenic Na pumping stimulated by Na entering with each impulse. Evidence is presented that the three effects are reversibly inhibited by conditions which depress active Na transport: (a) Li substituted for Na in the bath; (b) application of strophanthidin; (c) K removal; (d) treatment with cyanide; (e) cooling. We conclude that a single process is responsible for the three responses described above and identify that process as electrogenic Na pumping. Our observations also indicate that electrogenic pumping contributes to this neuron's resting potential.     

Adaptation in Cones: A General Model

Three features appear to characterize steady-state light adaptation in vertebrate cone photoreceptors: (a) the shape of the “log intensity-response” curve at different levels of adaptation is the same, the only change with adaptation is in the position of the point on the curve about which the cones operate; (b) at high adapting intensities the operating point becomes fixed in position; (c) this fixed position is at the steepest point of the log intensity-response curve. These three features can be described by a mathematical model.     

La conversion analogique numérique des récepteurs à l'étirement de la patte métathoracique du criquet Schistocerca gregaria

During the liarmonic stimulation of locust stretch-receptors of the metathoracic tibio-femoral joint, a decrease in the number of potentials in a sensory discharge can be observed with increasing frequency of stretching. On a log-log scale, a straight line of negative slope is calculated and fits best for each unit of both receptors (DPLR: dorso-postero-lateral receptor and DALR: dorso-antero-lateral receptor). These different graphs indicate that the number of impulses per cycle (N) decreases as a power function of the frequency of the sinusoidal input (F): logN = −alogF+b. For one unit, the slope a does not depend on the amplitude, the steady-state level (carrier rate: v), or temperature. The constant b gives a measure of the steady-state level at the start of stimulation, whereas a determines the dynamic sensitivity of the tested cell. When a unit is stimulated at the tonic frequency value of its steady-state level, only one spike is phase-locked to the stimulus, especially if the amplitude of the movement is large. Furthermore, when the frequency of stimulation is reduced to a half, the number of impulses per cycle doubles for the B and C units. With the method we have described here, one can hope to get further objective values to classify and to express the sensitivity of different kinds of mechanoreceptors.     

Ionic Control of the Reversal Response of Cilia in Paramecium caudatum : A calcium hypothesis

The duration of ciliary reversal of Paramecium caudatum in response to changes in external ionic factors was determined with various ionic compositions of both equilibration and stimulation media. The reversal response was found to occur when calcium ions bound by an inferred cellular cation exchange system were liberated in exchange for externally applied cations other than calcium. Factors which affect the duration of the response were (a) initial amount of calcium bound by the cation exchange system, (b) final amount of calcium bound by the system after equilibration with the stimulation medium, and (c) concentration of calcium ions in the stimulation medium. An empirical equation is presented which relates the duration of the response to these three factors. On the basis of these and previously published data, the following hypothesis is proposed for the mechanism underlying ciliary reversal in response to cationic stimulation: Ca⁺⁺ liberated from the cellular cation exchange system activates a contractile system which is energized by ATP. Contraction of this component results in the reversal of effective beat direction of cilia by a mechanism not yet understood. The duration of reversal in live paramecia is related to the time course of bound calcium release.     

Pacemaker Potentials for the Periodic Burst Discharge in the Heart Ganglion of a Stomatopod, Squilla oratoria

From somata of the pacemaker neurons in the Squilla heart ganglion, pacemaker potentials for the spontaneous periodic burst discharge are recorded with intracellular electrodes. The electrical activity is composed of slow potentials and superimposed spikes, and is divided into four types, which are: (a) "mammalian heart" type, (b) "slow generator" type, (c) "slow grower" type, and (d) "slow deficient" type. Since axons which are far from the somata do not produce slow potentials, the soma and dendrites must be where the slow potentials are generated. Hyperpolarization impedes generation of the slow potential, showing that it is an electrically excitable response. Membrane impedance increases on depolarization. Brief hyperpolarizing current can abolish the plateau but brief tetanic inhibitory fiber stimulation is more effective for the abolition. A single stimulus to the axon evokes the slow potential when the stimulus is applied some time after a previous burst. Repetitive stimuli to the axon are more effective in eliciting the slow potential, but the depolarization is not maintained on continuous stimulation. Synchronization of the slow potential among neurons is achieved by: (a) the electrotonic connections, with periodic change in resistance of the soma membrane, (b) active spread of the slow potential, and (c) synchronization through spikes.     

A Model Based on Receptor Desensitization for Cyclic AMP Signaling in Dictyostelium Cells

We analyze a model based on receptor modification for the cAMP signaling system that controls aggregation of the slime mold Dictyostelium discoideum after starvation. The model takes into account both the desensitization of the cAMP receptor by reversible phosphorylation and the activation of adenylate cyclase that follows binding of extracellular cAMP to the unmodified receptor. The dynamics of the signaling system is studied in terms of three variables, namely, intracellular and extracellular cAMP, and the fraction of receptor in active state. Using parameter values collected from experimental studies on cAMP signaling and receptor phosphorylation, we show that the model accounts qualitatively and, in a large measure, quantitatively for the various modes of dynamic behavior observed in the experiments: (a) autonomous oscillations of cAMP, (b) relay of suprathreshold cAMP pulses, i.e., excitability, characterized by both an absolute and a relative refractory period, and (c) adaptation to constant cAMP stimuli. A two-variable version of the model is used to demonstrate the link between excitability and oscillations by phase plane analysis. The response of the model to repetitive stimulation allows comprehension, in terms of receptor desensitization, of the role of periodic signaling in Dictyostelium and, more generally, the function of pulsatile patterns of hormone secretion.     

Most of the homoeologous pairing at metaphase I in wheat-rye hybrids is not chiasmatic

The use of telomeric C-bands in wheat-rye hybrids has made it possible to distinguish three types of wheat-wheat (1B^L) and wheat-rye associations (a, end-to-end extremely distal; b, end-to-ed distal; and c, interstitial) between homoeologous chromosomes at different metaphase I stages (early, middle and late) and also to estimate the actual recombination frequencies for such associations at anaphase I. There was a decrease of the a and b association frequencies during the different metaphase I stages, whereas the c type remained without variation in all stages. A good fit between the frequencies of c associations at metaphase I and the number of recombinant chromosomes at anaphase I, assuming a maximum of one chiasma per bond, was found; however, there was no correspondence between metaphase I and anaphase I data when all associations (a + b + c) were considered. In addition, rye-rye homologous pairing was observed at metaphase I, but no evidence for rye-rye recombination was found at anaphase I. The results indicate that most of end-to-end (a and b) homoeologous and nonhomologous associations are actually nonchiasmatic and are a remnant of prophase pairing.     

Dopamine release and molecular modeling study of some coumarin derivatives

4-aryl-2-amino-6-(4-hydroxy-2-oxo-2H-chromen-3-yl)-pyridin-3-carbonitrile (1), 4-aryl-2-oxo-6-(4-hydroxy-2-oxo-2H-chromen-3-yl)-pyridin-3-carbonitriles (2a-2c), 3-(6-aryl-1,2,5,6- tetrahydro-2-thioxopyrimidin-4-yl)-4-hydroxy-2H-chromen-2-one (3a, 3b) and pyrazol-3-yl-4-hydroxycoumarin derivatives (4a-4c, 5, 6a, 6b, 7a, 7b, and 8a-8c) were prepared in order to measure their % change dopamine release in comparison to amphetamine as reference, using PC-12 cells in different concentrations. In addition, the molecular modeling study of the compounds into 3BHH receptor was also demonstrated. The calculated inhibition constant (k_i) implemented in the AutoDock program revealed identical correlation with the experimental results to that obtained binding free energy (ΔG_b) as both parameters revealed reasonable correlation coefficients (R²) being 0.51 involving 10 compounds; (1, 2b, 2c, 3a, 3b, 4a, 4b, 6a, and 8c).     

Association between Psychopathic Disorder and Serum Antibody to Herpes Simplex Virus (Type 1)

The sera of a small of patients has been examined for herpes simplex virus antibody. Three clinically-defined groups of patients were compared: (a) aggressive psychopaths, (b) psychiatric controls, and (c) general hospital patients. The first group had an unusually high average kinetic neutralization constant against type 1 herpes simplex virus.     

The Kinetics of Distribution of the Fat-Soluble Inert Gas Cyclopropane in the Body

A kinetic analysis is made of the experimentally measured time course of respiratory uptake of the highly fat-soluble, inert gas cyclopropane by normal human subjects. The analysis is based on the well-known perfusion-limited model in which a number of body compartments are arranged in parallel with the lungs via the circulating blood. Three distinct body compartments are derived from the data. These are tentatively identified as: (a) adipose tissue (b) fat-poor tissue of low perfusion such as resting muscle, skin, and connective tissue (c) fat-poor tissue of high perfusion such as brain, heart, gut, liver, and kidney. Blood flow rates to the several compartments are also derived from the data. The rates to compartments (a) and (b) are each approximately 10 per cent of the estimated total cardiac output. The derived perfusion (blood flow rate/compartment weight) of the three compartments are in the range, respectively, (a) 2 to 4, (b) 1 to 2.5, (c) 25 to 75 ml/min/100 gm. Uncertainties arising from the experimental data and from simplifications of the model (neglect of lung fill-up phase of uptake and gross diffusion of cyclopropane from one tissue into another) are discussed. The present type of uptake experiment is significant for the problems of total body fat determination, of gross body composition in relation to weight change, of gross shunting of blood flow from one compartment to another, of anesthesia by fat-soluble substances, and of decompression sickness.     

The effects of air ions of the living mammalian trachea

Studies on the effects of air ions on the functional efficiency of the extirpated tracheal strip have been extended to the trachea of the living rabbit, rat, and mouse. Animals exposed to high mobility (+) air ions administered via a tracheotomy aperture displayed: (a) Decreased ciliary activity. (b) Decline in mucus flow rate, sometimes reversed by prolonged exposure to (+) ions; a frequent drop in the volume of mucous secretion. (c) Contraction of the membranous posterior tracheal wall. (d) Increased vulnerability to trauma of cilia and mucosal blood vessels. Similar treatment with (-) air ions reversed (+) ion effects on ciliary activity, mucus flow, contraction of the tracheal smooth muscle. Continued (-) ion treatment raised the ciliary rate (invariably) and the mucus flow rate (often) above their initial levels. (+) Air ions administered to unoperated resting mice and rats increased the respiratory rate; (-) ions reversed this effect. Long exposure of unoperated ambulatory mice to (+) air ions produced: (a) Decreased ciliary activity. (b) No clear cut effect on mucus flow. (c) Contraction of the posterior tracheal wall. (d) Increased vulnerability of the mucosa to trauma. (-) Air ions increased ciliary activity but had no clear-cut effect on the mucus flow rate.     

Phosphate-dependent Substrate Transport into Mitochondria: Oxidative Studies

The oxidation of malate and citrate by isolated plant mitochondria can be stimulated by the addition of inorganic phosphate. This stimulation (a) is not inhibited by oligomycin or uncouplers; (b) can not be duplicated by addition of adenine nucleotide; (c) is inhibited by 2-n-butylmalonate; and (d) is not evident in detergent-treated mitochondria. Phosphate was required to elicit uncoupler-stimulated respiration. It is concluded that these effects of phosphate are attributable to a stimulated rate of substrate penetration into the mitochondria, and do not involve the oxidative phosphorylation process.     

Diheteroarylmethyl substituted titanocenes: A novel class of possible anti-cancer drugs

From the reaction of tert-butyl lithium or n-butyl lithium with N-methylpyrrole (1a), furan (1b) or 2-bromo-thiophen (1c), 2-N-methylpyrrolyl lithium (2a), 2-furyl lithium (2b) or 2-thiophenyl lithium (2c), respectively, was obtained. When reacted with 6-(2-N-methylpyrrolyl) fulvene (3a), 6-(2-furyl) fulvene (3b) or 6-(2-thiophenyl) fulvene (3c), the corresponding lithiated intermediates were formed (4a-c). Titanocenes (5a-c) were obtained through transmetallation with titanium tetrachloride. When these titanocenes were tested against pig kidney epithelial (LLC-PK) cells, inhibitory concentrations (IC₅₀) of 32 μM, 140 μM, and 240 μM, respectively, were observed. These values represent improved cytotoxicity against LLC-PK, compared to their ansa-analogues.     

Analysis of the effects of calcium or magnesium on voltage-clamp currents in perfused squid axons bathed in solutions of high potassium

Isolated axons from the squid, Dosidicus gigas, were internally perfused with potassium fluoride solutions. Membrane currents were measured following step changes of membrane potential in a voltage-clamp arrangement with external isosmotic solution changes in the order: potassium-free artificial seawater; potassium chloride; potassium chloride containing 10, 25, 40 or 50, mM calcium or magnesium; and potassium-free artificial seawater. The following results suggest that the currents measured under voltage clamp with potassium outside and inside can be separated into two components and that one of them, the predominant one, is carried through the potassium system. (a) Outward currents in isosmotic potassium were strongly and reversibly reduced by tetraethylammonium chloride. (b) Without calcium or magnesium a progressive increase in the nontime-dependent component of the currents (leakage) occurred. (c) The restoration of calcium or magnesium within 15–30 min decreases this leakage. (d) With 50 mM divalent ions the steady-state current-voltage curve was nonlinear with negative resistance as observed in intact axons in isosmotic potassium. (e) The time-dependent components of the membrane currents were not clearly affected by calcium or magnesium. These results show a strong dependence of the leakage currents on external calcium or magnesium concentration but provide no support for the involvement of calcium or magnesium in the kinetics of the potassium system.     

ELECTROKINETIC PHENOMENA : VI. RELATIONSHIP BETWEEN ELECTRIC MOBILITY,CHARGE, AND TITRATION OF PROTEINS

1. By combining the theories of Smoluchowski, Debye and Hückel, and Henry it is possible to state explicitly (making necessary assumptions) under what conditions the following simple rule should be valid for proteins: In solutions of the same ionic strength, the electric mobilities of the same protein at different hydrogen ion activities should be proportional to the number of hydrogen (hydroxyl) ions bound. 2. Data of Tiselius and of the writer confirm this rule for (a) egg albumin, (b) serum albumin, (c) deaminized gelatin and gelatin, and (d) casein. 3. On the basis of the confirmed theory the titration curves of certain proteins are predicted from their mobilities. 4. It is shown that when certain proteins are adsorbed by quartz the apparent dissociation constant of the adsorbed protein is practically unchanged. The mass law must also be valid at the phase boundary. 5. The facts of paragraphs (1) to (4) are discussed in connection with the mechanism of (a) protein adsorption, (b) enzyme activity, (c) immune reactions, (d) the calculation of the electric charge of cells, and (e) criteria of surface similarity.     

Model of calcium movements in the mammalian myocardium: interval-strength relationship

A model is proposed to describe the interval-strength relationship in mammalian cardiac muscle in terms of "discrete" calcium movements associated with each cycle. The sarcoplasmic reticulum is assumed to be comprised of three functional sub-compartments: (1) The "main calcium store" which contains most of the calcium (predominantly bound) and is considered, due to its large buffering capacity, to account for the "long-term memory" lasting 7-10 beats. (2) The "releasable terminal" which contains the calcium readily available for release (all or most of it free) and accounts for the "short-term memory" which affects the subsequent beat. (3) The longitudinal network of the SR recirculating the myofibrillar calcium to the "main calcium store". The total content of calcium in the main store is determined by the transsarcolemmal influx and efflux. While influx occurs only during depolarization, efflux occurs during the whole cardiac cycle. The amount of free calcium in the main store is determined by an equilibrium equation. The release of calcium from the "releasable terminal" is governed by a "concentration-dependent" mechanism. This implies that when the concentration in the "releasable terminal" increases, the fraction released increases and the residual calcium left for the subsequent contraction decreases. The model predicts the following interval-strength relationships: steady state peak tension; changes from one steady rate to another; restitution curves; post-stimulation potentiation; paired stimulation; premature beats; post-extrasystolic potentiation following interpolated, basal or complimentary interval.