On the density of intracellular water

The density of individualArtemia cysts has been determined by sedimentation velocity measurements at unit gravity. Dried cyst (< 0.02 g H₂O/g dry weight) densities, _s were obtained by successive sedimentation in two nonpenetrating organic solvents. This removes geometric terms from the equation relating density to sedimentation velocity. Hydrated cysts ( 1.68 g H₂O/g dry weight) were sedimented in 0.0750 m NaCl to obtain their density ( _c). Values of _s, _c, and their ratios were found to be independent of cyst volume; therefore, the weight fraction of water in hydrated cysts is very nearly the same in cysts of greatly different size. It can be concluded that measurement of the water content of large populations of these cysts accurately reflects the water content of individual cysts, a point which has been assumed in previous work on this system. If _s does not change appreciably when dried cysts are fully hydrated then the density of their water, _w, can be calculated to be 1.022 g/cm³ (±0.0011 ). That value is significantly higher than the density of pure water and is very close to estimates of _w in skeletal muscle and amphibian oocytes obtained by others. However, the assumption that _s is independent of hydration is open to serious criticism, for all these studies. Consequently, conclusions and interpretations derived from such measurements must be considered to be tentative and uncertain.     

Modelling of predator–prey trophic interactions. Part I: two trophic levels

A class of lumped parameter models to describe the local dynamics in a controlled environment of a two-trophic chain is considered. The class is characterized by a trophic function (functional response of predator to the abundance of prey) depending on the ratio of prey biomass x and a linear function of predator biomass y: f(qx/[(1-)k+y]), where q is the efficiency of the predation process, k is a reference biomass, and (01) specifies the predation model. The trophic function is defined only by some properties determining its shape. A stability analysis of the models has been performed by taking the parameters q and as bifurcation parameters: the regions in the (,q) plane of existence and stability of nonnegative equilibrium states and limit cycles are determined. This analysis shows that the behaviour of the models is qualitatively similar for 0<1 (in particular the null state is always a saddle point), while the value =1 gives rise to some kind of structural instability of the system (in particular the null state becomes an attractor for sufficiently high predation efficiency).     

A mathematical theory of the functional dynamics of cortical and thalamic nervous tissue

It is proposed that distinct anatomical regions of cerebral cortex and of thalamic nuclei are functionally two-dimensional. On this view, the third (radial) dimension of cortical and thalamic structures is associated with a redundancy of circuits and functions so that reliable signal processing obtains in the presence of noisy or ambiguous stimuli.A mathematical model of simple cortical and thalamic nervous tissue is consequently developed, comprising two types of neurons (excitatory and inhibitory), homogeneously distributed in planar sheets, and interacting by way of recurrent lateral connexions. Following a discussion of certain anatomical and physiological restrictions on such interactions, numerical solutions of the relevant non-linear integro-differential equations are obtained. The results fall conveniently into three categories, each of which is postulated to correspond to a distinct type of tissue: sensory neo-cortex, archior prefrontal cortex, and thalamus.The different categories of solution are referred to as dynamical modes. The mode appropriate to thalamus involves a variety of non-linear oscillatory phenomena. That appropriate to archior prefrontal cortex is defined by the existence of spatially inhomogeneous stable steady states which retain contour information about prior stimuli. Finally, the mode appropriate to sensory neo-cortex involves active transient responses. It is shown that this particular mode reproduces some of the phenomenology of visual psychophysics, including spatial modulation transfer function determinations, certain metacontrast effects, and the spatial hysteresis phenomenon found in stereopsis.List of Symbols (t) Post-synaptic membrane potential (psp) - Maximum amplitude of psp - t Time - The neuronal membrane time constant - Threshold value of membrane potential - r Absolute refractory period - Synaptic operating delay - v Velocity of propagation of action potentil - x Cartesian coordinate - _jj (x) The probability that cells of class j are connected with cells of class j a distance x away - b _jj The mean synaptic weight of synapses of the jj-th class at x - _jj The space constant for connectivity - _e Surface density of excitatory neurons in a one-dimensional homogeneous and isotropic tissue - _i Surface density of inhibitory neurons in a one-dimensional homogeneous and isotropic tissue - E(x, t) Excitatory Activity, proportion of excitatory cells becoming active per unit time at the instant t, at the point x - I(x, t) Inhibitory Activity, proportion of inhibitory cells becoming active per unit time at the instant t, at the point x - x A small segment of tissue - t A small interval of time - P(x, t) Afferent excitation or inhibition to excitatory neurons - Q(x, t) Afferent excitation or inhibition to inhibitory neurons - N _e (x, t) Mean integrated excitation generated within excitatory neurons at x - N _i (x, t) Mean integrated excitation generated within inhibitory neurons at x - _e [N _e ] Expected proportion of excitatory neurons receiving at least threshold excitation per unit time, as a function of N _e - _i [N _i ] Expected proportion of inhibitory neurons receiving at least threshold excitation per unit time, as a function of N _i - G( _e ) Distribution function of excitatory neuronal thresholds - G( ₁ ) Distribution function of inhibitory neuronal thresholds - ₁ A fixed value of neuronal threshold - h(N _e ; ₁) Proportion per unit time of excitatory neurons at x reaching ₁ with a mean excitation N _e - 1[ ] Heaviside's step-function - R _e (x, t) Number of excitatory neurons which are sensitive at the instant t - R _i (x, t) Number of inhibitory neurons which are sensitive at the instant t - R _e Refractory period of excitatory neurons - r _i Refractory period of inhibitory neurons - E(x, t) Time coarse-grained excitatory activity - I(x, t) Time coarse-grained inhibitory activity - Spatial convolution - Threshold of a neuronal aggregate - v Sensitivity coefficient of response of a neuronal aggregate - E(t) Time coarse-grained spatially localised excitatory activity - I(t)> Time coarse-grained spatially localised inhibitory activity - L ₁,L ₂,L,Q See § 2.2.1, § 2.2.7, § 3.1 - Velocity with which retinal images are moved apart - Stimulus width - E _o, I _o Spatially homogeneous steady states of neuronal activity - k _e ,k _ij S _e S _ij See § 5.1     

A cone disk atomizer for production of biocompatible magnetic microcapsules for culture of anchorage-dependent mammalian cells

A cone disk atomizer has been designed for preparation of gelatin/polystyrene magnetic microcapsules for use as microcarriers in anchorage-dependent mammalian cell culture. This apparatus facilitates continuous production of microcapsules and ignores surfactants or organic solvents in traditional emulsion. Compared with the flat-disk atomizer, the feeding flow is subjected to a normal pressure provided by the special cone-shaped disk and liquid slide is dramatically decreased in the cone-disk atomizer. A flow-rate-limitation is observed in both flat-disk atomization and cone-disk atomization. A 36% increase in flow-rate limitation was observed in cone-disk atomization than in flat-disk atomization indicating the potential of this method for large-scale preparations. Nomenclature d _p average diameter of solid cores (m)D disk diameter (m)g gravity acceleration (9.8 m s^–2) Ga Galileo number defined by Equation (3) Q flow rate of feeding slurry (m³ s^–1) Re Q flow rate Reynolds number defined by Equation (2)Re rotating speed Reynolds number defined by Equation (1) angular speed of rotating disk (rad s^–1) _l density of liquid (kg m^–3) _s density of the mixing slurry (kg m^–3) density difference between solid cores and liquid (kg m^–3) viscosity of the mixing slurry (Pa s)     

Measurement of proton relaxation rates in proteins

Summary Five different types of experiment are described which make it possible to measure various relaxation rates of selected protons in crowded spectra of macromolecules such as proteins: longitudinal spin-lattice relaxation rates =1/T₁, transverse relaxation rates =1/T₂ measured under conditions of free precession, transverse relaxation rates ¹ _LOCK=1/T₁ measured under conditions of spin-locking, and transverse relaxation rates ^DQC=1/T₂ ^DQC and ^ZQC=1/T₂ ^ZQC of double- and zero-quantum coherences. The surprisingly large discrepancy between the transverse rates ^t and ^t is discussed in detail. To separate overlapping proton signals, the experimental schemes involve one or several magnetization transfer steps, using a doubly selective homonuclear Hartmann-Hahn method. Numerous variants of the basic ideas can be conceived, depending on the extent of signal overlap and on the topology of the networks of scalar couplings. Applications are shown to H and H of Tyr²³, to H, H and H of Cys³⁰, and to H and H of Ala²⁴ in bovine pancreatic trypsin inhibitor (BPTI).     

Globally asymptotic properties of proliferating cell populations

This paper presents a general model for the cell division cycle in a population of cells. Three hypotheses are used: (1) There is a substance (mitogen) produced by cells which is necessary for mitosis; (2) The probability of mitosis is a function of mitogen levels; and (3) At mitosis each daughter cell receives exactly one-half of the mitogen present in the mother cell. With these hypotheses we derive expressions for the and curves, the distributions of mitogen and cell cycle times, and the correlation coefficients between mother-daughter (_md) and sister-sister (_ss) cell cycle times.The distribution of mitogen levels is shown to be given by the solution to an integral equation, and under very mild assumptions we prove that this distribution is globally asymptotically stable. We further show that the limiting logarithmic slopes of (t) and (t) are equal and constant, and that _md0 while _ss0. These results are in accord with the experimental results in many different cell lines. Further, the transition probability model of the cell cycle is shown to be a simple special case of the model presented here.     

Alternative pathway activities in aged potato tuber slices measured by three methods

To find a simple and reliable oxygen electrode-based method to estimate the values of alternative pathway activity (V _alt) and its contribution to total respiration V _alt/V _t) in aged potato (Solanum tuberosum L.) tuber slices, we compared conventional hydroxamate-inhibiting method, improved hydroxamate-inhibiting method with 2,6-dichlorophenol indophenol (DCPIP), and the oxygen isotope discrimination (OID) method. The values of V _alt and V _alt/V _t obtained with an improved hydroxamate-inhibiting method with DCPIP in 12-h- and 24-h-aged slices were about twice higher than those with the conventional hydroxamate-inhibiting method. Only a relatively small difference in the values of V _alt and V _alt/V _t obtained by the OID method and the improved hydroxamate-inhibiting method with DCPIP in 12-h and 24-h-aged slices was observed. These results indicated that the improved hydroxamate-inhibiting method with DCPIP could be considered as a new, simple, and reliable technique for the noninvasive assay of the AP activity.From Fiziologiya Rastenii, Vol. 52, No. 2, 2005, pp. 311–315.Original English Text Copyright © 2005 by Hou, Zhou, Kong, Liang, Zhang.This article was submitted by the authors in English.This revised version was published online in April 2005 with a corrected cover date.     

Interaction of Phomopsin A with Normal and Subtilisin-Treated Bovine Brain Tubulin

Tubulin, the major component of microtubules, has a tendency to lose its ability to assemble or to bind to ligands in a time-dependent process known as decay. The decay process also causes tubulin to expose sulfhydryl groups and hydrophobic areas. The antimitotic drug phomopsin A strongly protects the tubulin molecule from decay. Here we have studied the interaction of phomopsin A with tubulin and tubulin which has been treated with subtilisin to remove selectively the C-termini of the and chains (_ss). The binding of phomopsin A to tubulin decreases the sulfhydryl titer by approximately 1.0 mol/mol. Selective removal of the peptides from the C-terminal ends does not affect phomopsin A's interaction with tubulin. Moreover, the _ss tubulin–phomopsin A complex appears to be more stable than the tubulin–phomopsin A complex as determined by the time-dependent increase in exposure of sulfhydryl groups and hydrophobic areas on tubulin. In fact, phomopsin A inhibits the decay process of _ss tubulin completely. This observation raises the possibility of determining the conformtion of this configuration of tubulin.     

Rotational diffusion tensor of nucleic acids from 13C NMR relaxation

Rotational diffusion properties have been derived for the DNA dodecamer d(CGCGAATTCGCG)₂ from ¹³C R₁ and R₁ measurements on the C₁, C₃, and C₄ carbons in samples uniformly enriched in ¹³C. The narrow range of C-H bond vector orientations relative to the DNA axis make the analysis particularly sensitive to small structural deviations. As a result, the R₁/R₁ ratios are found to fit poorly to the crystal structures of this dodecamer, but well to a recent solution NMR structure, determined in liquid crystalline media, even though globally the structures are quite similar. A fit of the R₁/R₁ ratios to the solution structure is optimal for an axially symmetric rotational diffusion model, with a diffusion anisotropy, D_||/D, of 2.1±0.4, and an overall rotational correlation time, (2D_||+4D)^–1, of 3.35 ns at 35 °C in D₂O, in excellent agreement with values obtained from hydrodynamic modeling.     

The taxonomic significance of the trunk limbs of the chydoridae (Cladocera)

Summary The differences in the structure of the trunk limbs allow to outline three sections of Chydoridae (see table I and fig. 1), coinciding with the sections distinguished according to the structure of the head pores.

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Chydoridae (Cladocera)

Chydoridae (. ), , .

     

Bacteriophage phi 1 as a gene-cloning vector in Bacillus subtilis

Summary We attempted to use Bacillus subtilis phage 1 as a gene-cloning vector since the 1 genome was found to have few cleavage sites upon digestion with several kinds of restriction endonucleases. A 1 stock supplied by J. Ito (University of Arizona, Tucson, USA) consisted of two phages, 1E1 and 1E2, having one and two EcoRI-cleavage sites in their genomes respectively. From the latter isolate a deletion mutant 1E21 was induced to increase the size range of DNA segments to be cloned. It was demonstrated, by in vitro recombination experiments with phage 11 DNA, that 1E21 can be used for cloning EcoRI fragments of various sizes. We analyzed the DNAs of ten 1 clones isolated from independent transfectants and found that six of them carried 11 DNA fragments inserted at either of the two EcoRI-cleavage sites. Some of the hybrid phage DNAs were found to be cleaved with BamHI and HaeIII endonucleases at the 11 DNA portion, whereas the parental 1E21 DNA was insensitive to any of these enzymes. These hybrid phages would therefore be useful vectors for cloning foreign DNA fragments generated by cleavage with BamHI or HaeIII endonucleases.     

Changes of fluorescence lifetime and rotational correlation time of bovine serum albumin labeled with 1-dimethylaminonaphthalene-5-sulfonyl chloride in guanidine and thermal denaturations

The nanosecond fluorescence depolarization method was applied to measure the fluorescence lifetime () and the rotational correlation time () of bovine serum albumin (BSA) labeled with 1-dimethylaminonaphthalene-5-sulfonyl chloride (dansyl-Cl). Changes of and of dansyl BSA in the guanidine denaturation and in the thermal denaturation were examined. In parallel, the secondary structural change of dansyl BSA was followed by circular dichroism measurements. The magnitude of was almost unchanged between 1 and 2 M guanidine, where the secondary structure of the protein was predominantly disrupted; whereas that of began to increase before the disruption of secondary structure in the guanidine denaturation. In the thermal denaturation, in contrast, changes of both and occurred in a temperature range where the secondary structure was predominantly disrupted. The volume of equivalent sphere (V _e ) and the axial ratio () for the BSA were 3.6–3.8×10^–19 cm³ and 3.6 at 2M guanidine as against 2.1×10^–19 cm³ and 2.2 in the absence of guanidine (25°C), respectively. The magnitudes ofV _e and were 4.9×10^–19 cm³ and 4.5 at 65°C, respectively. Although the secondary structural change of dansyl BSA was irreversible in the thermal denaturation,V _e and were reversible.     

Die Aktivit?tsperiodik bei V?geln mit und ohne dunklem Schlafkasten

Zusammenfassung In 2 Versuchsserien wurden Kohlmeisen(Parus major) und japanische Möwchen(Lonchura striata var.domestica) einzeln und schallisoliert gehalten. In der ersten Versuchsserie, in der alle Vögel einen dunklen Schlafkasten hatten, wurde der Einfluß der Beleuchtungsstärke auf die Periode () der Hüpfaktivität und auf das Verhältnis von Aktivitätszeit zu Ruhezeit ( : -Verhältnis) untersucht. Sowhol Kohlmeisen als auch japanische Möwchen folgen der Regel, daß mit wachsender Beleuchtungsstärke die Periode kürzer und das : -Verhältnis größer wird.In der 2. Serie wurde der Einfluß Ruhe im dunklen Schlafkasten auf die Periodenlänge und auf das : -Verhältnis untersucht. Es wurden die Messungen aus Bedingung 1 (der Vogel hat einen dunklen Schlafkasten zur Verfügung) mit den Messungen aus Bedingung 2 (der Vogel hat keinen oder einen hellen Schlafkasten zur Verfügung) verglichen. Das Ergebnis bei Kohlmeisen entspricht den Befunden bei konstantem Licht verschiedener Intensität. Unter Bedingung 1 ist länger und das : -Verhältnis kleiner als in Bedingung 2. Das Ausmaß der Änderung von nach Fortnahme des dunklen Kastens ist unabhängig von der Periodenlänge in Bedingung 1. Das Ausmaß der änderung von : ist unabhängig von a : in Bedingung 1, jedoch schwach negativ korreliert mit der Periodenlänge in Bedingung 1.Bei japanischen Möwchen entsprechen die Ergebnisse dieser Versuchsserie nicht der Regel für tagaktive Vögel. Mit Benützen des dunklen Schlafkastens ist kürzer als ohne den Schlafkasten. Ohne den Schlafkasten ist etwa 24 Std. Das : -Verhältnis ist in Bedingung 1 unter bestimmten Voraussetzungen kleiner als in Bedingung 2. Das Ausmaß der Änderung von nach Fortnahme des Kastens ist mit der dazugehörigen Periode in Bedingung 1 hochsignifikant korreliert (Regressionskoeffizient b=-1.01, Korrelationskoeffizient r=0.89). Ebenfalls ist das Ausmaß der Änderung von : nach Fortnahme des Kastens mit : aus Bedingung 1 korreliert; es scheint, als würde ein bevorzugtes : -Verhältnis von etwa 2.0 eingeregelt.Die Ergebnisse werden im Hinblick auf 4 Punkte diskutiert: 1) Das circadiane System arbeitet innerhalb eines engen Bereiches von - und : -Werten optimal. 2) Der Optimalbereich wird bevorzugt unter ungünstigen Bedingungen angestrebt. 3) Der Entzug des dunklen Schlafkastens belastet japanische Möwchen mehr als Kohlmeisen. 4) Bei japanischen Möwchen wird in Bedingung 1 durch fortplfanzungsphysiologischen Einfluß verkürzt.

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Circadian activity rhythms of birds with and without a dark nest box

Summary Perch-hopping activity of Great tits(Parus major) and Bengales finches(Lonchura striata domestica), housed individually in soundproof boxes, was studied in two series of experiments. In the first series all birds had access to a dark nest box, in which they retired during their subjective night. In this experiment the effect of light intensity on the freerunning circadian activity rhythm was investigated. Both Great tits and Bengalese finches obey the circadian rule by responding to an increase in light intensity with shortening the circadian period () and with an increase of the ratio of activity time and rest time ( : ).In the second series of experiments the influence of sleeping in the dark nest box on both circadian period and : -ratio was studied. The results of two experimental conditions — without and with access to a dark nest box — were compared. In the Great tits, the results are in agreement with the effect of light intensity: when a dark nestbox is available, is longer and the : -ratio is smaller than in the absence of a nest box. The magnitude of the change in free-running period after removal of the nest box is independent of the original value of ; the amount of change : -ratio is likewise independent of the original : -ratio, but is weakly correlated to the original .InLonchura striata var. domestica, removal of the dark nest box leads to a lenghtening of the free-running period to about 24 hours; the : -ratio is smaller in the presence of a dark nestbox, if certain other conditions are fulfilled. The magnitude of the change in after removal of the nest box is highly correlated to the original free-running period (r=-0.89) in such a way that, without nest box, the period approaches a value of 24 hours. Also, the amount of change in : -ratio due to nest box removal is negatively correlated to the original : -ratio. A probably preferred : -ratio of 2.0 is adopted.These results are discussed in the view of 4 points: 1) The circadian system operates at its optimum within a narrow range of - and : -values. 2) This optimal range is especially adopted when conditions become adverse. 3) Removal of the dark nest box results in a more stressful situation for Bengalese finches than for Great tits. 4) In the Bengalese finches, is shortened in the presence of a nest box due to effects on reproductive physiology.

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Herrn Prof. Dr. JürgenAschoff zum 60. Geburtstag gewidemt.     

Sodium channel opening as a precursor to inactivation

The time constant of the process producing the delay in Na inactivation development as determined by the two pulse method (_delay) was extracted and compared to that of the slowest Na activation process ₃ for the I _Na during the conditioning pulse of that same determination. _delay and two pulse inactivation _c values were computer generated using a nonlinear least squares algorithm. _h and single pulse inactivation _h values were independently generated for each determination also with the aid of the computer using the same non-linear least squares algorithm. In one determination at 2 mV, _c was 4.68 and _delay 0.494 ms while _h was 4.70 and ₃ 0.491 ms for a _c/_h of 0.996 and a _delay/₃ of 1.006. Mean _delay/₃ from five determinations in four axons, both Cs and K perfused, and spanning a potential range of-27 to 2mV was 1.068. The precursor process to inactivation is channel opening. Some fraction of channels presumably inactivate via another route where prior channel opening is not required.     

Variable generation times and Darwinian fitness measures

Summary Reproductive value (RV) and net reproductive output (R _o) are frequently used fitness measures. We argue that they are only appropriate when intervals between reproductive events are fixed, as they are dimensionless generation-to-generation scalings with units offspring per parent. A fitness measure should account for two different effects of a decrease in generation time: (1) increased survival due to shorter exposure to mortality agents and (2) increased frequency of reproduction.R _o andRV deal with the first of these two effects, while a measure with a physical dimensionper time [T^–1] is needed to account for the second. The Malthusian growth parameter,r, meets this requirement and in situations where time to reproduction is variable, we propose, the instantaneous rate of spread of descendants (from an individual) be used instead ofR _o. As an alternative toRV, we suggest using the instantaneous difference = –r, wherer is the population rate of increase. WhileRV andR _o are dimensionless ratios, , and areper time rates which are appropriate in accounting for alterations in generation time.     

Substitutions of the Conserved Thr42 Increased the Roles of the ε-Subunit of Maize CF1 as CF1 Inhibitor and Proton Gate

The conserved residue Thr42 of -subunit of the chloroplast ATP synthase of maize (Zea mays L.) was substituted with Cys, Arg, and Ile, respectively, through site-directed mutagenesis. The over-expressed and refolded -proteins were purified by chromatography on DEAE-cellulose and FPLC on mono-Q column, which were as biologically active (inhibiting Ca²⁺-ATPase activity and blocking proton gate) as the native subunit isolated from chloroplasts. The T42C and T42R showed higher inhibitory activities on the soluble CF₁(–) Ca²⁺-ATPase than the WT. The T42I inhibited the Ca²⁺-ATPase activity of soluble CF₁ and restored photophosphorylation activity of membrane-bound CF₁ deficient in the most efficiently. Far-ultraviolet CD spectra showed that the portions of -helix and -sheet structures of the three mutants were somewhat different from WT. Thus the conserved residue Thr42 may be important for maintaining the structure and function of the -subunit and the basic functions of the -subunit as far as an inhibitor of Ca²⁺-ATPase and the proton gate are related.     

A method for the estimation of gene flow parameters from a population structure caused by restricted gene flow and genetic drift

Summary A method has been developed which enables the estimation of the plant gene flow parameters _p (pollen dispersal), _s (seed dispersal) and t (outcrossing rate) from a selection-free continuously structured population in equilibrium. The method uses Wright's F-coefficients and introduces a new F-function which describes the genetic similarity as a function of the spatial distance. The method has been elaborated for wind pollinated plant species but can be modified for insect pollination and for animal species. In practice allozymes will provide for the necessary neutral genetic variation. The more loci used and the more intermediate the gene frequencies, the more reliable the results. For the estimation of _p and t together (when the outcrossing rate is not known) at least two chromosomally unlinked loci are required. The method for estimating _s depends on whether the plant species is annual or perennial. The mechanism of selfing has been analysed by the explanation of the value of t by three components: population density (d), pollen flow (_p) and relative fertilization potential of own pollen (Z). The concepts of neighbourhood size and isolation by distance, developed by Wright, who used a single gene flow parameter , have been extended to the situation which is realistic for seed plants, using all three parameters _p, _s and t. When _p is large with respect to _s, _s largely determines the value of the neighbourhood size, whereas _p is the most dominating factor in isolation by distance. The use of local effective population size and mean gene transport per generation instead of neighbourhood size and neighbourhood area, respectively, is proposed to avoid confusion. Computer simulations have been carried out to check the validity and the reliability of the method. Populations of 200 plants, using two or three loci with intermediate allele frequencies, gave good results in the calculation of _p with known value of t and of _s and N_e. With unknown t, especially with lower values of t, larger populations of at least 1,000 plants are necessary to obtain reasonably accurate results for _p and mean gene transport per generation M.Grassland Species Research Group Publication No. 81     

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.     

Identification of Slow Motions in the Reduced Recombinant High-potential Iron Sulfur Protein I (HiPIP I) from Ectothiorhodospira Halophila via 15N Rotating-frame NMR Relaxation Measurements

Rotating-frame 15N relaxation rate (R1) NMR experiments have been performed in order to study the dynamic behavior of the reduced recombinant high-potential iron-sulfur protein iso I (HiPIP I) from Ectothiorhodospira halophila, in the s to ms time range. Measurements of R1 were performed as a function of the effective spin-lock magnetic field amplitude by using both on and off-resonance radio frequency irradiation. The two data sets provided consistent results and were fit globally in order to identify possible exchange processes in an external loop of the reduced HiPIP I. The loop consists of residues 43-45 and the correlation time of the exchange process was determined to be 50 ± 8 s for the backbone nitrogen of Gln 44.     

Inhibition of DNA-ethidium bromide intercalation due to free radical attack upon DNA

Summary The fluorescent intercalation complex of ethidium bromide (ETB) with DNA was used as a probe to compare the effects of various radicals with respect to impairment of the DNA base-pair region.OH radicals inhibit up to 0.7 dye intercalations perOH at low salt concentration, and for various oxidizing species the effect decreases in the orderOH > Br ₂ ^– > N ₃ > $$ " align="middle" border="0"> (SCN) ₂ ^–. DNA impairment by theOH product of Met-Gly is comparable to that of N ₃ , but no effect was found due to the interaction between DNA and Lys-Tyr-Lys phenoxyl radicals. The reducing speciese _aq ^– , H, O ₂ ^–, and CO ₂ ^– hardly affect the DNA-ETB intercalation.