Effects of zwitterionic detergents on the electronic structure of the primary donor and the charge recombination kinetics of P+Q A − in native and mutant reaction centers from Rhodobacter sphaeroides

The effects of detergents on the electronic structure of the oxidized primary donor P⁺ and the time constant _AP of the P⁺Q _A ^– charge recombination at ambient temperatures have been investigated in native and mutant reaction centers (RCs) from Rhodobacter sphaeroides. It is shown that N-lauryl-N,N-dimethyl-3-ammonio-1-propane sulfonate (SB12) induces a transition to a second distinct conformation of the RC. In the case of the wild type and the mutant FY(M197), in which a hydrogen bond is introduced to the 2-acetyl group of the dimer half of P that is associated with the M-subunit of the RC, the conformational change causes a more asymmetric spin density distribution between the two bacteriochlorophyll moieties of P⁺ in favor of the L-half. For both types of RCs the time constant _AP depends on the SB12/RC ratio as does the position of the long-wavelength band of P, _max. The increase of _AP by 30 ms and the shift of _max from 866 nm to 851 nm are indicative for the conformational change. In addition, a smaller linear increase of _AP with increasing SB12/RC ratio is superimposed on the variation of _AP due to the conformational change. Similar effects of SB12 on the optical spectra as well as on _AP are also observed for the two heterodimer mutants HL(L173) and HL(M202), in which one of the bacteriochlorophylls of P is replaced by a bacteriopheophytin. There are no clear indications for a correlation of _AP with the localization of the positive charge in P⁺. Furthermore, it is concluded from the dependence of _AP on the SB12/RC ratio that the single-site mutations do not affect the standard free energy difference of the two conformations to a measurable extent.     

Effect of axoplasmic transport blockade on end-plate currents in frog muscle fibers

Evoked and spontaneous end-plate currents (EPC) were studied in normal voltage-clamped frog sartorius muscle fibers and 2 weeks after application of colchicine to the nerve innervating the muscle to block axoplasmic transport in its fibers. Application of colchicine was found to reduce the rate of rise and to prolong decay of EPC without affecting the amplitude of the EPC and miniature EPC, the quantum composition of EPC, and the frequency of miniature EPC. The histogram of distribution of the time constant () of EPC decay under normal conditions follows the normal law, but after application of colchicine to the nerve it is shifted to the right, with separation of two modes (₁ and ₂). Three types of synapses can be distinguished from the character of EPC decay: monoexponential decay with ₁ (44%), biexponential decay with ₁ and ₂ (39%), and monoexponential decay with ₂ (19%). An increase in of EPC decay is accompanied by strengthening of the dependence of this process on the clamping voltage. The current-voltage characteristic and reversal potential of EPC are unchanged. It is suggested that the change in character of EPC decay after application of colchicine to the motor nerve is due to the appearance of acetylcholine-activated ionic channels in the muscle membrane with a longer duration of the open state and with potential-dependence of the open state similar to that taking place after muscle denervation.S. V. Kurashov Medical Institute, Ministry of Health of the RSFSR, Kazan'. Translated from Neirofiziologiya, Vol. 17, No. 2, pp. 204–211, March–April, 1985.     

Potential induced in a spherical cell by an intracellular point source and an extracellular point sink

Summary The potential is calculated for all time, inside and outside a spherical cell for a point source of current inside the cell and a point sink located a finite distance outside the cell. The source and sink are step functions in time. An eigenfunction expansion is obtained, valid for arbitrary =_m a/_i , where _i and _m are the conductivities inside the cell and in the membrane, respectively, a is the cell radius and the membrane thickness. For small , the eigenfunction expansion is expanded in powers of . The time dependence of the potential contains transients with two widely differing time constants =C_m a/_i, where C_m is the membrane surface capacitance, and _m=/. Closed-form expressions are obtained for the two leading terms, for small , after the rapid transient is over. The remaining time dependence is only in the potential inside the cell, and is a simple exponential increase, independent of position within the cell. It is found that the transmembrane potential is insensitive to the location of the extracellular sink at long times, but not at short times. The dependence of the potential on location of source, sink, and observer is studied for long times after the quick transients are over. A uniqueness theorem is derived for the solution to Laplace's equation for the membrane boundary condition.This work was supported in part by NSF Grant No. GB-24965. Dr. Peskoff is the recipient of NIH Special Research Fellowship No. 1F03 GM 55849-01. Mr. Ramirez is a Ford Foundation Pre-doctoral Fellow.     

Application of the Criterion of Biological Time (τ n /τ0) to the Studies of Embryogenesis in Birds

It was shown on the example of chick embryo that the number ₀ ( _n /₀) can be recommended as a measure of biological time and, for this purpose, the duration of the minimal mitotic cycle during synchronous cleavage divisions should be determined (in minutes) in various avian species. Based on the preliminary data, one can propose the comparability and similarity of the temporal programs of gastrulation and somitogenesis in the chick embryo and embryos of some fish and amphibians.     

Photocurrent kinetics of purple-membrane sheets bound to planar bilayer membranes

Summary The kinetics of light-driven proton transport by bacteriorhodopsin has been studied in a model system consisting of a planar lipid bilayer membrane to which purple membrane fragments have been attached. After excitation with a 10-nsec laser flash a fast negative current-transient occurs, followed by a positive transient which decays to zero. The time course of the photocurrent may be represented by a sum of four exponentials with time constants ₁= 1.2sec, ₂= 17sec, ₄= 57sec, ₁= 950sec (at 25°C). In a D₂O medium ₂ and ₃ are increased by a factor of 2.6 and 2.9, respectively, whereas ₁ remains unaffected. The observed components of the photocurrent can be correlated to photochemical reaction steps inferred from flash-photometric experiments on the basis of the observed time constants, the activation energies, and the effects of pH and D₂O. From the photocurrent signals information may be obtained on the magnitude of the charge displacement associated with the elementary transitions of the bacteriorhodopsin molecule.     

Effective diffusivity and tortuosity in a porous glass immobilization matrix

The effective diffusivity of glucose in porous glass beads was determined using a transient method. Predictions for the intraparticle and surface concentrations were made by an analytical solution of the mass balance. The value of the diffusivity was expected to be lower than the value of the corresponding diffusion coefficient in water, but the opposite was observed. This effect results from intraparticle fluid flow, leading to high values of the apparent effective glucose diffusivity. To measure diffusion only and to prevent any internal convection during the diffusion experiment, the pores of the porous glass beads were filled with Ca-alginate gel. For these glass beads (internal porosity, , equal to 0.56), we found an effective glucose diffusivity of 2.2×10^–10 m²/s at 30°C. Using the relationship to effective intraparticle diffusivity (D_eff)=effective diffusivity in 1% Ca-alginate beads (D_gel) / (with the tortuosity factor) this gives =1.7. For known and measuring by the method described, the D_eff can be calculated for other porous materials or diffusing substances. Knowledge of the exact value of the effective diffusivity is a necessity in bioreactor modelling and was demonstrated by prediction of the residence time distribution profiles in a packed-bed bioreactor containing immobilized yeast cells.     

Holling's “hungry mantid” model for the invertebrate functional response considered as a Markov process. Part I: The full model and some of its limits

In this paper we give an analytical reformulation of Holling's (1966) simulation model for invertebrate predatory behaviour. To this end we represent a population of predators as a frequency distribution over a space of (physiological) states. The functional response of a predator is calculated from the (stable) equilibrium distribution of its state as a function of prey density.Starting from the general model various other models are obtained by limit processes, some of them new and some of them old. The more interesting of which will be studied in further papers in this series.List of Notation a rate constant of digestion - b maximum of rate constant of prey encounter in the mantid - b maximum pursuit duration in the mantid (_p(0)) - c satiation threshold for search - c satiation threshold for pursuit in the mantid: c=c(b-D_s/v)/b - D _m maximum sighting distance - D _p pursuit distance - D _s strike distance - expectation operator - f, f ₀ rate of change of satiation during search - f ₁ rate of change of satiation during prey handling - F functional response: number of prey eaten per unit of time by one predator - g rate constant of effective prey encounter in the gobbler and sucker - g₀ rate constant of prey encounter - g₁ probability of no prey loss from pursuit - g₂ probability of no prey escaping during pursuit - H Holling secretary correction factor in the sucker: fraction of the time spent searching - k _R density of R - k_T probability density of maximum prey handling time - K probability that maximum prey handling time is _e, i.e. pursuit duration is zero - K _R distribution function of R - N number of prey caught - p (marginal) density of S - p₀ density of S in search - p₁ simultaneous density of S and T - P probability - p ₁ marginal density of S in handling prey - q probability of strike success - R ratio of realized to maximum sighting distance - s, S satiation - satiation axis - t time - handling time axis - u eating speed - U homogeneous(0,1) random variable - v pursuit speed - V exponential(1) random variable - w prey weight - W exponential(_m) random variable - x prey density - ratio of maximum successful pursuit duration to meal duration (_pm/_e) - _pm - relative duration of successful pursuit (_p/_pm) - ratio of shortest to largest sighting distance - x_e - time already spent handling a prey item - rate of prey loss during prey handling - prey escape rate during pursuit - prey biomass density (xw) - , T maximum time still to be spent handling a prey item - _e meal duration - _m maximum handling time ( _e+ _p) - _p duration of successful pursuit - _pm maximum duration of successful pursuit (_p(0)) - hazard rate - _m maximum of hazard rate - scaled functional response (wF) - minimal i-state space     

Conformational dynamics of the anticodon loop in yeast tRNAPhe as sensed by the fluorescence of wybutine

Conformational and dynamic properties of the anticodon loop of yeast tRNA^Phe were investigated by analyzing the time resolved fluorescence of wybutine serving as a local structural probe adjacent to the anticodon GmAA on its 3 side. The influence of Mg²⁺, important for stabilizing the tertiary structure of tRNA, and of the complementary anticodon s²UUC of E. coli tRNA ₂ ^Glu were investigated.Fluorescence lifetimes and anisotropies were measured with ps time resolution using time correlated single photon counting and a mode locked synchronously pumped and frequency doubled dye laser as excitation source. From the analysis of lifetimes () and rotational relaxation times ( _R ) we conclude that wybutine occurs in various structural states: (i) one stacked conformation where the base has no free mobility and the only rotational motion reflects the mobility of the whole tRNA molecule (=6 ns, _R =19 ns), (ii) an unstacked conformation where the base can freely rotate (=100 ps, _R = 370 ps) and (iii) an intermediary state (=2 ns, _R = 1.6 ns).Under biological conditions, i. e. in the presence of Mg²⁺ and neutral salts, wybutine is found in a stacked and immobile state which is consistent with the crystallographic picture. In the presence of the complementary codon however, as exemplified by the E. coli-tRNA ₂ ^Glu anticodon, our analysis indicates that the codon-anticodon complex exists in an equilibrium of structural states with different rotational mobility of wybutine. The conformation with wybutine freely mobile is the predominant one and suggests that this conformation of the codon-anticodon structure differs from the canonical 3–5 stack.     

Electrical characteristics of sensory neurons of Hirudo medicinalis

During intracellular polarization of identified sensory neurons of the leech by square pulses of hyperpolarizing current electrical parameters of the cell membranes were determined: input resistance of the neuron R_n, time constant of the membrane , the ratio between conductance of the cell processes and conductance of the soma , the resistance of the soma membrane r_s, the input resistance of the axon r _a , capacitance of the membrane C_s, and resistivity of the soma membrane R_s. The results obtained by the study of various types of neurons were subjected to statistical analysis and compared with each other. Significant differences for neurons of N- and T-types were found only between the values of , C_s, and R_s (P<0.01). These parameters also had the lowest coefficients of variation. The surface area of the soma of the neurons, calculated from the capacitance of the membrane (the specific capacitance of the membrane was taken as 1 µF/cm²) was 7–10 times (N-neurons) or 4–6 times (T-neurons) greater than the surface area of a sphere of the same diameter. The resistivity of the soma membrane R_s was 35.00 k·cm² for cells of the N-type and 19.50 k·cm² for T-neurons. The reasons for the relative stability of this parameter compared with the input resistance of the cell (coefficient of variation 22–7 and 53–31% respectively) are discussed. The possible effects of electrical characteristics on the properties of repeated discharges in neurons of different types also are discussed.A. A. Zhdanov Leningrad State University. Translated from Neirofiziologiya, Vol.7, No.3, pp.295–301, May–June, 1975.     

Effects of moisture stress on the multiplication and expansion of cells in leaves of sugar beet

Summary Sugar beets were subjected to moisture stress by decreasing the water potential of the culture solution osmotically with polyethylene glycol by a known amount, , and, alternatively by applying matric potential, , at the plant roots. Lowering the water potential at the root surface less than 200 millibars by either method resulted in significant decreases in the rate of cell multiplication. The final number of cells per leaf at = -372 mb the final was 165% of that at = -473 mb ( = –101 mb); similarly at = –15 mb the final cell number was 198% of that at = –196 mb ( = –181 mb). The mean cell volume of leaves was not significantly affected by these levels of moisture stress.     

Modelling endplate currents: dependence on quantum secretion probability and decay of miniature current

Quantification of the time course and amplitude of endplate currents (EPC) was made with respect to dispersion of quanta secretion and to changes in the exponential decay of miniature endplate currents (_mepc). The relationship between RPC amplitude and _mepc follows a double-exponential curve with ₁= 0.3 ms and ₂ = 6 ms. If the amplitude of fully synchronised EPC is taken as 100%, then the loss of EPC amplitude is already 42% with physiological parameters of dispersion (the half-rise and decay constant of distribution of secretion probability = 0.5 ms, _mepc =1 ms). This loss is even more substantial if secretion is more dispersed or miniature endplate currents decay faster. Correspondence to: F. Vyskocil     

Photodynamic action inSpirostomum ambiguum

Summary The photodynamic action inSpirostomum ambiguum has been studied measuring the time lag between a flash of visible light and the induced contraction of spirostomum. This quantity shows an exponential dependence on the flash strength and on the dye concentration. These measurements indicate that there is in spirostomum a natural content of photodynamic substances and that there is a natural rhythm of contraction of about 900 sec (at the temperature of 20 °C) even in darkness and without added photodynamic substances. Also the temperature dependence of has an exponential behaviour with an activation energy of 0.37 eV.The effect of the partial pressure of oxygen on has also been investigated and a hyperbolic dependence is shown to occur. The action of the oxygen on the photodynamic effect can be described by a relationship which is similar to that employed to describe the biological damage by X-rays.The results are analyzed in terms of a model which accounts for the action of two kinds of molecules activated by light: molecules which are always active (n ₂) and molecules which are active only when reacting with oxygen (n ₁). This model suggests that the photodynamic action in spirostomum is regulated by an autocatalytic process.     

"Velocity leakage" in the pigeon vestibulo-ocular reflex

The transfer characteristics of the vestibuloocular reflex (VOR), and of the semicircular canal primary afferents (SCPAs) that drive it, have been studied in several species. In monkeys and cats, the dominant time constant describing horizontal VOR dynamics ( _hu ) is longer than that ( _c ) of horizontal SCPAs. This lengthening of the time constant has been attributed to a velocity storage mechanism that has been modeled as a positive feedback loop in the VOR pathways. We have studied the transfer characteristics of horizontal and vertical VOR and SCPAs in unanesthetized pigeons. In this species the dominant time constants of both the horizontal and vertical VOR ( _hv and _vv ) are shorter than _c . This finding indicates that time constants characterizing the lower frequency response of the VOR can be lengthened or shortened depending on the species. We propose that in the pigeon the velocity leakage mechanism can be modeled by substituting negative feedback for positive feedback in the model of the VOR pathways. Negative feedback can also account for the further shortening of _hu and _vv as VOR gain increases with arousal. Additionally, making the negative feedback loop nonlinear can model the dependency of lower frequency VOR phase on amplitude, and skew in VOR waveforms. Pigeon VOR and SCPA dynamics also differ in their adaptive properties and higher frequency behavior. A predominance of input from highly adaptive SCPAs is proposed to account for the increased adaptation of the vertical VOR as compared with SCPAs overall. A pure time-delay associated with VOR operation can explain the phase lag of the VOR relative to SCPAs at higher frequencies.     

Dispersal distance of corn pollen under fluctuating diffusion coefficient

I develop a mechanistic model for pollen dispersal by a generalization of the Brownian motion model. This model provides an explanation as to why the distribution of the dispersal distance is leptokurtic in most organisms. The pollen is assumed to move in a period between and + to the circumference of a circle of radius r that has a center at the current position. The angle of movement is chosen at random. Unlike the conventional Brownian motion model, the step length, r, fluctuates in a stochastic manner, obeying a generalized gamma distribution. The convection flow, i.e., the directional movement of pollen, is also considered. I show that this model is identical to the diffusion model where the dispersal duration weighted by the diffusion coefficient obeys a gamma distribution. Hence, the model is called the gamma model. The solution is given in an explicit form. The model is fitted to six data sets obtained from the literature by maximizing the quasi-likelihood. Another model (the inverse gamma model), which is not a mechanistic model but a phenomenological model, is also fitted to the data to evaluate the validity of the gamma model.     

Existence of a non-constant periodic solution of a non-linear autonomous functional differential equation representing the growth of a single species population

Summary We consider an integro-differential equation for the densityn of a single species population where the birth rate is constant and the death rate depends on the values ofn in an interval of length — 1 > 0. We prove the existence of a non-constant periodic solution under the conditions birth rate b > /2 and - 1 small enough. The basic idea of proof (due to R. D. Nussbaum) is to employ a theorem about non-ejective fixed points for a translation operator associated with the solutions of the equation.A proof of existence was also announced by G. Dunkel in [1].     

Growth analysis of the self-thinning stands of Pinus densiflora Sieb. et Zucc

The competition-density (C-D) effect for self-thinning Pinus densiflora Sieb. et Zucc. stands was analyzed. The relationship between biological time and physical time t followed a hyperbolic curve. The coefficients A_t and B included in the reciprocal equation of the C-D effect in self-thinning stands (i.e. 1/w=A_t+B), where w and , respectively, represent the mean stem volume and the realized stand density, were calculated at each time. With increasing , the coefficient A_t increased abruptly up to a maximum value, and then decreased gradually to a constant level, whereas the coefficient B decreased exponentially. The relationship between the realized stand density and the initial stand density _i was confirmed to follow the equation: 1/=1/_i+, where 1/ represents the asymptotic stand density at a given time. The - relationship was represented by the equation: =p(e–1), where p and are constants. The density in the self-thinning stands tended to converge to the same density level after a sufficient lapse of time, irrespective of the difference in initial stand density. The time-trajectory of the mean stem volume and asymptotic stand density on logarithmic coordinates moved gradually toward the self-thinning line with a slope of approximately –3/2, whereas the time-trajectory of the mean stem volume and full stand density moved initially along the self-thinning line with a slope of approximately –3/2, and then changed to move along the maximum yield line with a slope of –1.0.     

Activation kinetics of single high-threshold calcium channels in the membrane of sensory neurons from mouse embryos

Summary Activation kinetics of single high-threshold inactivating (HTI orN-type) calcium channels of cultured dorsal root ganglion cells from mouse embryos was studied using a patchclamp method. Calcium channels displayed bursting activity. The open-time histogram was single exponential with an almost potential-independent mean open time _op. The closed-time histogram was multicomponent; at least three of the components were associated with the activation process. The fast exponential component with the potential-independent time constant _cl ^f included all intraburst gaps, while two slower ones with potential-dependent time constants _cl ^vs described shut times between bursts and between clusters of bursts. The burst length histogram was biexponential. The fast component with a relatively potential-independent time constant _bur ^f described short, isolated channel openings while the slow component characterized real bursts with a potential-dependent mean life time. The waiting-time histogram could be fitted by a difference of two exponentials with time constants being the same as _cl ^s and _cl ^vs . The data obtained were described in the frame of a 4-state sequential model of calcium channel activation, in which the first two stages are formally attributed to potential-dependent transmembrane transfer of two charged gating particles accompanying the channel transitions between three closed states, and the third one to fast conformational changes in channel protein leading to the opening of the channel. The rate constants for all transitions were defined. The validity of the proposed model for both low-threshold inactivating (LTI orT-type) and high-threshold noninactivating (HTN orL-type) calcium channels is discussed.     

Responsiveness of cardiac Na+ channels to antiarrhythmic drugs: The role of inactivation

Summary Elementary Na⁺ currents were recorded at 9°C in inside-out patches from cultured neonatal rat heart myocytes. In characterizing the sensitivity of cooled, slowly inactivating cardiac Na⁺ channels to several antiarrhythmic drugs including propafenone, lidocaine and quinidine, the study aimed to define the role of Na⁺ inactivation for open channel blockade.In concentrations (1–10 mol/liter) effective to depressNP _o significantly, propafenone completely failed to influence the open state of slowly inactivating Na⁺ channels. With 1 mol/liter, _open changed insignificantly to 96±7% of the control. Even a small number of ultralong openings of 6 msec or longer exceeding _open of the whole ensemble several-fold and attaining _open (at –45 mV) in cooled, (-)-DPI-modified, noninactivating Na⁺ channels proved to be drug resistant and could not be flicker-blocked by 10 mol/liter propafenone. The same drug concentration induced in(-)-DPI-modified Na⁺ channels a discrete block with association and dissociation rate constants of 16.1 ± 5.3 × 10⁶ mol^–1 sec^–1 and 675 ± 25 sec^–1, respectively. Quinidine, known to have a considerable affinity for activated Na⁺ channels, in lower concentrations (5 mol/liter) left _open unchanged or reduced, in higher concentrations (10 mol/liter) _open only slightly to 81% of the predrug value whereasNP _o declined to 30%, but repetitive blocking events during the conducting state could never be observed. Basically the same drug resistance of the open state was seen in cardiac Na⁺ channels whose open-state kinetics had been modulated by the cytoplasmic presence of F^– ions. But in this case, propafenone reduced reopening and selectively abolished a long-lasting open state. This drug action is unlikely related to the inhibitory effect onNP _o since hyperpolarization and the accompanying block attenuation did not restore the channel kinetics. It is concluded that cardiac Na⁺ channels cannot be flicker-blocked by antiarrhythmic drugs unless Na⁺ inactivation is removed.     

Energy migration in purple bacteria. The criterion for discrimination between migration- and trapping-limited photosynthetic units

A criterion has been evolved for distinguishing between migration- and trapping-limited photosynthetic units (PSUs). Its application to purple bacteria has proved their PSUs to be of trapping-limited type. It means that any improvements of the molecular structure of their PSUs cannot noticeably increase the overall rate constant of excitation delivery from antenna BChls to reaction centers (RCs).Abbreviations PSUs photosynthetic units - RCs reaction centers - Chl chlorophyll - BChl bacteriochlorophyll - R intermolecular distance, _e - quantum yields of the primary excitation trapping and wasteful losses respectively - _fl excitation and fluorescence lifetimes respectively     

Inactivation of calcium currents in the soma of spinal ganglia by removing the membrane potential depolarization shift

Kinetic and voltage-dependent characteristics of the inactivating action of incoming calcium currents were investigated in the somatic membrane of rat spinal ganglia neurons using an intracellular dialysis technique. It was shown that the "tail" of low-threshold calcium current could be reliably described by one exponent with a time constant of =1.2–1.8 msec at a repolarization potential of –90 mV. The "tail" of the high-threshold calcium current represented the sum of several exponents. The time constant of the main component which expressed inactivation of the high threshold calcium current was ^h=250–350 µsec. It was also shown that and ^h remained virtually unchanged for repolarization potentials in the subthreshold region; they increase, however, if the repolarizing potential is close to those potentials at which the corresponding component of calcium current is initially activated. A dependence was observed between the levels and ^h and duration of the depolarizing shift. Findings are discussed in the context of a three-tier model of calcium channels.A. A. Bogomolets Institute of Physiology, Academy of Sciences of the Ukrainian SSR, Kiev. Translated from Neirofiziologiya, Vol. 17, No. 5, pp. 682–691, September–October, 1985.