Effects of black widow spider venom and Ca2+ on quantal secretion at the frog neuromuscular junction

A modification of the classical procedure of fluctuation analysis is used to measure the waveform, w(t), mean amplitude, (h), and mean rate of occurrence, (r), of miniature endplate potentials (MEPPs) at frog cutaneous pectoris neuromuscular junctions treated with black widow spider venom (BWSV). MEPP parameters are determined from the power spectrum of the fluctuating potential and the second (variance), third (skew), and fourth semi-invariants (cumulants) of high-pass-filtered records of the potential. The method gives valid results even when the mean potential undergoes slow changes unrelated to MEPPs and when the MEPP rate is not stationary; it detects changes in the distribution of MEPP amplitudes and corrects for the nonlinear summation of MEPPs. The effects of Ca2+ on BWSV-induced secretion are studied in detail. When Ca2+ is absent, the power spectrum of the fluctuations is shaped like the spectrum of w(t) and secretion is quasi-stationary; (r) rises smoothly to peak values of approximately 1,500/s and then quickly subsides to levels near 10/s. Many relatively small and some "giant" MEPPs occur at the ends of the experiments, and the distribution of MEPP amplitudes broadens. When the effects of this broadening are corrected for, we find that approximately 0.7 X 10(6) MEPPs occurred during the 30 min of intense secretion. Since BWSV depletes nerve terminals of their quanta of transmitter and their synaptic vesicles, this figure is an upper limit for the quantal store in a resting terminal. When Ca2+ is present, the noise spectrum deviates from the spectrum of w(t) and secretion is nonstationary; (r) rises to similar peak values but is sustained at levels near 400/s for up to an hour and at least 1.5 X 10(6) quanta are secreted within this period. Thus, the quantal store must have turned over at least twice under this condition. Data previously obtained at junctions treated with La3+ are corrected for nonlinear summation and for the distribution of MEPP amplitudes. The two corrections roughly compensate each other, and the corrected results confirm the previous conclusion that the number of quanta secreted from La3+-treated terminals during 1 h is not strongly dependent upon the extracellular concentration of Ca2+; approximately 2 X 10(6) quanta are released even when Ca2+ is absent.     

Basketball Shot Types and Shot Success in Different Levels of Competitive Basketball

The purpose of our research was to investigate the relative frequencies of different types of basketball shots (above head, hook shot, layup, dunk, tip-in), some details about their technical execution (one-legged, two-legged, drive, cut, …), and shot success in different levels of basketball competitions. We analysed video footage and categorized 5024 basketball shots from 40 basketball games and 5 different levels of competitive basketball (National Basketball Association (NBA), Euroleague, Slovenian 1^st Division, and two Youth basketball competitions). Statistical analysis with hierarchical multinomial logistic regression models reveals that there are substantial differences between competitions. However, most differences decrease or disappear entirely after we adjust for differences in situations that arise in different competitions (shot location, player type, and attacks in transition). Differences after adjustment are mostly between the Senior and Youth competitions: more shots executed jumping or standing on one leg, more uncategorised shot types, and more dribbling or cutting to the basket in the Youth competitions, which can all be attributed to lesser technical and physical ability of developing basketball players. The two discernible differences within the Senior competitions are that, in the NBA, dunks are more frequent and hook shots are less frequent compared to European basketball, which can be attributed to better athleticism of NBA players. The effect situational variables have on shot types and shot success are found to be very similar for all competitions.     

Fluorescence Decay Time Distribution Analysis Reveals Two Types of Binding Sites for 1,8-Anilinonaphthalene Sulfonate in Native Human Oxyhemoglobin

Binding of 1,8-anilinonaphthalene sulfonate (1,8-ANS) with native human oxyhemoglobin (Hb) in 50 mM potassium phosphate buffer (pH 7.4) was studied by steady state fluorescence spectroscopy and by laser spectrofluorimetry with subnanosecond time resolution. The distribution of fluorescence decay times and parameters of two- and three-exponential deconvolution of the fluorescence kinetics of 1,8-ANS in Hb solution demonstrate that the emission at wavelengths _em of 455-600 nm is not single-exponential and has components with mean decay times <0.5, 3.1-5.5, and 12.4-15.1 nsec with the amplitudes depending on the emission wavelength. Analysis of time-resolved fluorescence spectra shows that the shortest-lived component should be assigned to 1,8-ANS molecules in the aqueous medium, whereas the two longer-lived components are assigned to two types of binding sites for 1,8-ANS in the Hb molecule characterized by different polarity and accessibility to water molecules.     

在体大鼠海马CA1区侧枝/联合纤维通路和TA通路的不同EPSP空间整合特性

在麻醉Wistar大鼠上,结合脑室给药,应用双电极刺激技术刺激海马独立的两条侧枝／联合纤维通路、TA通路,并在CA1区放射层记录兴奋性突触后电位（EPSP）,对海马CA1区锥体细胞近、远端树突EPSP的空间整合进行了初步探讨。结果表明,海马CA1区锥体细胞近、远端树突的空间整合都是亚线性的;近端树突的空间整合不受期望值大小的影响,但远端树突的空间整合随期望值增加而减小（更趋于亚线性）。此外,荷包牡丹碱没有影响EPSP的空间整合;但瞬时A型钾通道（IAK^＋）的拮抗剂氨基吡啶-4却使得近端树突的空间整合趋于线性发展。本研究表明,海马CA1锥体细胞近、远端树突不同的被动、主动特征使它们具有了不同的空间整合特性。由于近端树突接受海马内部侧枝／联合纤维投射的信息,远端树突通过TA通路接受内嗅皮层投射的信息,由此提示,CA1区锥体细胞对来自海马内部和直接来自皮层的信息输入采用了不同的整合方式。     

A general rule for sensory cue summation: evidence from photographic, musical, phonetic and cross-modal stimuli

The Euclidean and MAX metrics have been widely used to model cue summation psychophysically and computationally. Both rules happen to be special cases of a more general Minkowski summation rule , where m = 2 and ∞, respectively. In vision research, Minkowski summation with power m = 3-4 has been shown to be a superior model of how subthreshold components sum to give an overall detection threshold. Recently, we have previously reported that Minkowski summation with power m = 2.84 accurately models summation of suprathreshold visual cues in photographs. In four suprathreshold discrimination experiments, we confirm the previous findings with new visual stimuli and extend the applicability of this rule to cue combination in auditory stimuli (musical sequences and phonetic utterances, where m = 2.95 and 2.54, respectively) and cross-modal stimuli (m = 2.56). In all cases, Minkowski summation with power m = 2.5-3 outperforms the Euclidean and MAX operator models. We propose that this reflects the summation of neuronal responses that are not entirely independent but which show some correlation in their magnitudes. Our findings are consistent with electrophysiological research that demonstrates signal correlations (r = 0.1-0.2) between sensory neurons when these are presented with natural stimuli.     

Correlations among parameters that describe low-spin ferric heme complexes

The g values from low-spin ferric hemes can be related through the t2g hole model to rhombic (V/lambda) and tetragonal (delta/lambda) ligand field components and to the lowest Kramer's doublet energy (E/lambda). The latter is also a measure of unpaired electron sharing among the iron 3d (t2g) orbitals. For a series of ligands (X), there is a monotonic increase in myoglobin complex (Mb . X) [E/lambda] values with nonheme hexacoordinate metal complex (M . X6) [eg-t2gPg] orbital separations. As the aqueous solution pKa values of the sulfurous or nitrogenous ligands in model heme complexes increase, values of V/lambda and delta/lambda increase linearly, but those of [E/lambda] decrease linearly. The greater the electron-acceptor ability of the ligand, as suggested by its position in the spectrochemical series or its pKa, the more the unpaired electron sharing among the heme t2g orbitals increases. The rate of change of [E/lambda] with V/lambda and the pKa is different with sulfurous and nitrogenous ligands, and the magnitude of both rates increases with two sulfurs less than sulfur and nitrogen less than two nitrogens bound to the heme. The maximum magnitude of this rate with V/lambda for cytochrome P-450 is four times less than that for myoglobin, which may explain, in part, the differences in ligand binding between these two hemeproteins. The perturbation of [E/lambda], V/lambda, and delta/lambda induced by strain of iron-ligand bonds is quantitated for several hemeproteins and heme models. In addition, energy level comparisons suggest that the largest-magnitude g value falls approximately along the iron-chlorin ring normal. This suggestion implies that the electron distribution of the iron at the catalytic sites of cytochrome P-450 and certain chlorin-containing enzymes is in some way similar, but distinct from that at the transport site of myoglobin.     

Effects of isolated and combined exposures to whole-body vibration and noise on auditory-event related brain potentials and psychophysical assessment

Auditory event-related brain potentials (ERP) in response to two different tone stimuli (1.1 kHz or 1 kHz, 80 dB, 50 ms; given by headphones at a regular interstimulus interval of 5 s with a probability distribution of 70:30) were recorded from 12 healthy male subjects (Ss) during four different conditions with two repetitions: A-60 dBA white noise (wN), no whole-body vibration (WBV); B-60 dBA wN plus sinusoidal WBV in the az-direction with a frequency of 2.01 Hz and acceleration of 2 m.s-2 root mean square; C-80 dBA wN, no WBV; D-80 dBA wN plus WBV. Each condition consisted of two runs of about 11 min interrupted by a break of 4 min. During the break with continuing exposure, but without auditory stimuli, Ss judged the difficulty of the tone-detection task and intensity of noise by means of cross-modality matching (CMM). Vibration-synchronous activity in the electrocardiogram was eliminated by a subtraction-technique. Noise caused an attenuation of the N1 and P2 amplitudes and prolongation of P3 latencies. The WBV did not cause systematic ERP effects. Condition B was associated with higher N1 and smaller P3 amplitudes. The factor "condition" had a significant effect on the peak latencies of P3 to target stimuli and the task difficulty judged by CMM. Both effects exhibited significant linear increases in the sequence of conditions A, B, C, D. For the evaluation of exposure conditions at work, it can be suggested that noise has a strong systematic effect which can be enhanced by WBV.(ABSTRACT TRUNCATED AT 250 WORDS)     

Response histogram shapes and tuning curves: The predicted responses of several cortical cell types to drifting gratings stimuli

The responses to visual stimuli of simple cortical cells show linear spatial summation within and between their receptive field subunits. Complex cortical cells do not show this linearity. We analyzed the simulated responses to drifting sinusoidal grating stimuli of simple and of several types of complex cells. The complex cells, whose responses are seen to be half-wave rectified before pooling, have receptive fields consisting of two or more DOG (difference-of-Gaussians) shaped subunits. In both cases of stimulation by contrast-reversal gratings or drifting gratings, the cells' response as a function of spatial frequency is affected by the subunit distances 2 and the stimulation frequency . Furthermore, an increased number of subunits (a larger receptive field) yields a narrower peak tuning curve with decreased modulation depth for many of the spatial frequencies. The average and the peak response tuning curves are compared for the different receptive field types.     

Interspike Interval Fluctuations in the Crayfish Stretch Receptor

Trains of nerve impulses from the crayfish stretch receptor under steady conditions are found to be extremely regular with a standard deviation of S = 0.7 x 10(-2)tau when tau, the mean interval, is in the 15 to 80 msec range. Such a small fluctuation increases the problems of film recording, accurate measurement of intervals, and especially, drifts. Experimental and mathematical techniques are described to obviate these problems. Evidence is found for a serial correlation coefficient of about -0.2 in the range, tau = 35 msec to tau = 70 msec. Shot noise and Johnson noise in a long axon are evaluated in detail and are shown to be comparable in size. It is also shown that neither shot noise nor Johnson noise is large enough to explain simply the observed interval fluctuations. Other types of membrane noise are discussed.     

The equivalence of fluctuation analysis and chemical relaxation measurements: a kinetic study of ion pore formation in thin lipid membranes

(1) Autocorrelation measurements were made of the current fluctuations due to ion-conducting channels produced by gramicidin A in black lipid membranes. (2) Relaxation experiments using a voltage jump were made on the same system. (3) Reciprocal time constants were determined by both methods over a 10⁸ fold range in membrane conductivity. starting from the single channel level. (4) The mean squared amplitudes of the fluctuations were determined from the autocorrelation functions. (5) The data were tentatively rationalized on the basis of a hypothetical dimerization reaction, assuming that gramicidin A dimers form conducting channels. The same forward rate constants are obtained, via (3), by both methods (1) and (2). The backward rate constant agrees excellently with direct measurements of the mean life time of a conducting channel. (6) The unit channel conductance and - assuming again a dimerization - the equilibrium constant can be obtained from the fluctuation amplitude distribution.     

Determination of Pre- and post-tetanic quantum values in excitatory synapses of CA1 area of hippocampal slices: Application of the deconvolution method

The deconvolution method, i.e., reconstruction of noise-free discrete amplitude distributions of excitatory postsynaptic potentials (EPSPs), has been tested in computer experiments. A generalized quantal model without constraints on distribution of transmitter release probabilities (p) was used. Complex binomial amplitude distributions with a number of discrete components n between 2 and 10 and with different p were simulated. The distance between the discrete components (i.e., the quantum value, v) was determined from simulated distributions with the number of amplitudes N equal to 1000 or 100. For samples with N=100, mean v values were calculated from 10 distributions. The solutions obtained by deconvolution were close to (within ±20% of) the simulated v values when the standard deviation of the noise Sn0.5v. For Sn=0.5–2.0v, v grew in proportion to Sn and was overestimated. A similar correlation was found for v calculated from amplitudes of minimal EPSPs recorded from CA1 area neurons of guinea pig hippocampal slices. However, no significant correlation between v and Sn was found for v>135 µV, i.e., for estimates exceeding the total mean value. A substantial increase in the mean quantal content (m) with a relatively slight increase in v during longterm potentiation (LTP) was confirmed for those cases. Methods of elimination of erroneous v estimates were considered. Rejection of cases with v>2.5Sn (when N=1000) or v>3Sn (when N=100) eliminated erroneous estimates with a high probability but demanded very low noise levels. A new criterion for recognition of correct estimates is suggested. It is based on artificial contamination of the measured amplitudes by the Gaussian noise (noise addition) and on the double-step dependence of v on Sn described above. The solutions selected with this criterion also support a predominant increase of m during LTP with a slight growth of v.Translated from Neirofiziologiya, Vol. 25, No. 2, pp. 84–91, March–April, 1993.     

Lesions in thighs from hunted Brown Hares (<Emphasis Type="Italic">Lepus europaeus</Emphasis>) and microflora under vacuum-packaging storage

In two surveys, thighs of a total of 137 hunted hares were tested for the presence of intramuscular shots and femur fractures, which were detected in 42.7% and 29.2% of 274 thighs, respectively. Femur fractures were significantly associated with the presence of intramuscular shots. In the second survey (46 hares), 92 thighs were grouped into three categories, “A” (no fractures, no intramuscular shot), “B” (one intramuscular shot), and “C” (multiple shots and hematoma), with 49.0%, 33.6%, and 17.4%, respectively. Category “C” was found unfit for human consumption. During 7-day storage of vacuum-packed “A” and “B” thighs, total aerobic counts increased from initially 3.3 ± 0.3 (mean ± SD) and 4.1 ± 0.6 log cfu/g by ca. 2 log units when stored at 3–4°C, whereas the increase was clearly <1 log unit at 0°C. In comparison to temperature, differences between “A” and “B” category were less pronounced. Similar dynamics were observed for Enterobacteriaceae. In all categories, muscle pH values (mean = 5.83) were similar. It is concluded that storage at temperatures of ca. 4°C, although in compliance with EU legislation, does not afford keeping microbial contaminants in check, and thus will not preserve microbiological quality of vacuum-packed hare meat.     

The problem of shot selection in basketball

In basketball, every time the offense produces a shot opportunity the player with the ball must decide whether the shot is worth taking. In this article, I explore the question of when a team should shoot and when they should pass up the shot by considering a simple theoretical model of the shot selection process, in which the quality of shot opportunities generated by the offense is assumed to fall randomly within a uniform distribution. Within this model I derive an answer to the question “how likely must the shot be to go in before the player should take it?” and I show that this lower cutoff for shot quality depends crucially on the number of shot opportunities remaining (say, before the shot clock expires), with larger demanding that only higher-quality shots should be taken. The function is also derived in the presence of a finite turnover rate and used to predict the shooting rate of an optimal-shooting team as a function of time. The theoretical prediction for the optimal shooting rate is compared to data from the National Basketball Association (NBA). The comparison highlights some limitations of the theoretical model, while also suggesting that NBA teams may be overly reluctant to shoot the ball early in the shot clock.     

Electrophysiological correlates preceding and following the movement onset in man

The cortical potential changes associated with unilateral voluntary self-paced hand movements were detected over the surface of the scalp by the summation method of EEG activity in 20 young subjects. A typical complex wave form of average movement potential (AMP): N1, P1, N2, P2, were discerned in all subjects in our records. This paper presents the results of the topographical distribution of the second potential of the AMP (Premotion Positivity, P1) and the last potential of the AMP (Positive Postmovement onset Potential, P2). Our results indicate a bilateral symmetrical presence of both positive components precentrally and parietally. They also indicate that both these potentials are bilaterally large posterior to the rolandic fissure, and laterality effects in amplitudes occurred only in the second positive wave parietally during right-hand responses in right handers.     

Timescales of population rarity and commonness in random environments

This is a mathematical study of the interactions between non-linear feedback (density dependence) and uncorrelated random noise in the dynamics of unstructured populations. The stochastic non-linear dynamics are generally complex, even when the deterministic skeleton possesses a stable equilibrium. There are three critical factors of the stochastic non-linear dynamics; whether the intrinsic population growth rate (lambda) is smaller than, equal to, or greater than 1; the pattern of density dependence at very low and very high densities; and whether the noise distribution has exponential moments or not. If lambda < 1, the population process is generally transient with escape towards extinction. When lambda > or = 1, our quantitative analysis of stochastic non-linear dynamics focuses on characterizing the time spent by the population at very low density (rarity), or at high abundance (commonness), or in extreme states (rarity or commonness). When lambda >1 and density dependence is strong at high density, the population process is recurrent: any range of density is reached (almost surely) in finite time. The law of time to escape from extremes has a heavy, polynomial tail that we compute precisely, which contrasts with the thin tail of the laws of rarity and commonness. Thus, even when lambda is close to one, the population will persistently experience wide fluctuations between states of rarity and commonness. When lambda = 1 and density dependence is weak at low density, rarity follows a universal power law with exponent -3/2. We provide some mathematical support for the numerical conjecture [Ferriere, R., Cazelles, B., 1999. Universal power laws govern intermittent rarity in communities of interacting species. Ecology 80, 1505-1521.] that the -3/2 power law generally approximates the law of rarity of 'weakly invading' species with lambda values close to one. Some preliminary results for the dynamics of multispecific systems are presented.     

The effect of binocular stimulation on each component of transient and steady-state VEPs

We recorded the monocular and binocular VEPs to the alternation of sinusoidal gratings in order to evaluate the binocular interaction in each component of transient and steady-state VEPs in 13 normal subjects. Three spatial frequencies (1.3, 2.6 and 5.3 c/deg) with a 90% contrast were used as visual stimuli. The latencies and amplitudes of N70 and P100 of the transient VEPs were measured. The steady-state VEPs were Fourier analyzed, and both the phase and amplitude of the second (2F) and fourth (4F) harmonic responses were obtained. Binocular interaction was influenced by spatial frequency such that a binocular summation or even an inhibition occurred. For the transient VEPs, a binocular summation was more pronounced in the amplitude of N70 than in that of P100 at all spatial frequencies. There were no significant effects of binocular stimulation on latencies of N70 or P100. However, the latencies of N70 and P100 showed different spatial frequency characteristics. For the steady-state VEPs, the amplitude of 2F revealed a binocular summation that was more pronounced at 5.3 c/deg, whereas the 4F amplitude showed binocular inhibition at 2.6 and 5.3 c/deg. The 2F phase showed binocular inhibition at all spatial frequencies, whereas no such inhibition was observed in the 4F phase. These results suggest that individual components of transient and steady-state VEPs are physiologically distinct and may therefore be generated from different neuronal populations in striate cortex.     

Switch to non-toxic shot in the Camargue,France: effect on waterbird contamination and hunter effectiveness

The use of lead shot for wildfowling is a severe threat to waterbirds through ingestion of toxic lead pellets. Consequently, lead shot has been banned in many countries since the late 1990s and on Tour du Valat estate since 1994 (Camargue, France). An experimental study was undertaken to check if hunters would habituate to this new type of cartridge and consequently improve their effectiveness (average number of shots per bagged animal), assess the factors influencing effectiveness, and assess the trend of contamination in shot waterbirds and the estate sediment. From 1995 to 2005, we monitored hunting bags, spent cartridges, and the gizzards of shot ducks. Using generalized mixed effect models, we assessed the factors influencing hunter effectiveness. Instances of non-toxic pellets in duck gizzards increased, probably as a result of rapid accumulation in the sediments. We estimated that between 1995 and 2005, the lead shot ban spared 456 kg of lead from entering 403 ha of temporary marshes and avoided the contamination of 8 % of the ducks foraging on Tour du Valat. After 11 years of hunting with non-toxic shot, there was unexpectedly no clear pattern in trends of individual effectiveness among hunters. Hunter effectiveness was instead positively influenced by game abundance and hunter assiduity and negatively influenced by wind and number of shots, suggesting a lassitude effect. Our results suggest crippling loss can be reduced through regular practice, self-limitation of shooting intensity to below 20 shots to avoid lassitude effects, and self-limitation of shooting distance under strong wind conditions.     

Properties of miniature postsynaptic currents during depolarization-induced release at a cholinergic neuroneuronal synapse

1. Miniature postsynaptic currents were analyzed at an inhibitory cholinergic neuroneuronal synapse in the buccal ganglion of Aplysia. Under double voltage-clamp, it was possible to induce postsynaptic currents by long-duration depolarizations of the presynaptic neuron and to analyze these as the linear summation of individual miniature postsynaptic currents (MPSCs). The amplitude of these miniature currents (imin) was calculated from the ratio of the variance of the noise (E2) to the mean of the postsynaptic current (Im), according to Campbell's theorem, with imin = 2E2/Im. Their decay time (tau min) was obtained from the cutoff frequencies of the power spectra obtained from the noise. 2. Neither the conductance nor the decay time of MPSCs was voltage dependent. However, imin appeared to decrease when the quantal content of the response increased. Meanwhile, tau min increased slightly with Imin. 3. Carbamylcholine was injected into the neuropile and this led to a decrease in imin and a slight increase in tau min. 4. Power spectra obtained after the application of inhibitors of acetylcholinesterase (AChE), with or without curare, suggested that acetylcholine (ACh) does not accumulate during large depolarizations. 5. The possible origin of the nonlinear relationship between the variance and the mean of the postsynaptic currents is discussed.     

Fabrication of Gold Nanostructures and Studies of Their Morphological and Surface Plasmonic Properties

We report fabrication of gold nanostructures on glass and indium tin oxide (ITO)-coated glass substrates using high fluence and highly energetic gold ions generated by hot, dense, and strongly non-equilibrium plasma. Nanodots and nanorods are observed in scanning electron microscopy (SEM) of nanostructures grown on glass substrate with single and double shots of gold ions which is in conformity with the transmission electron microscopy image. SEM images for single and double shots of gold ions on ITO-coated glass substrate show only nanodots. The mean diameter of nanodots obtained on both glass and ITO-coated glass is found to increase with increase in the number of gold ions shot from one to two. The gold nanostructures exhibit red shift in surface plasmon resonance with increased interaction which is in agreement with other reported work.     

Folding kinetics of phage 434 Cro protein

Folding kinetics for phage 434 Cro protein are examined and compared with those reported for lambda(6-85), the N-terminal domain of the repressor of phage lambda. The two proteins have similar all-helical structures consisting of five helices but different stabilities. In contrast to lambda(6-85), sharp and distinct aromatic (1)H NMR signals without exchange broadening characterize the native and urea-denatured 434 Cro forms at equilibrium at 20 degrees C, indicating slow interconversion on the NMR time scale. Stopped-flow fluorescence data using the single 434 Cro tryptophan indicate strongly urea-dependent refolding rates and smaller urea dependencies of the unfolding rates, suggesting a native-like transition state ensemble. Refolding rates are slower and unfolding rates considerably faster at pH 4 than at pH 6. This accounts for the lower stability of 434 Cro at pH 4 and suggests the existence of pH-dependent, possibly salt bridge interactions that are more stabilizing at pH 6. At <2 M urea, decreased folding amplitudes and nonlinear urea dependencies that are apparent at pH 6 indicate deviation from two-state behavior and suggest the formation of an early folding intermediate. The folding behavior of 434 Cro and why it folds 2 orders of magnitude slower than lambda(6-85) are rationalized in terms of the lower intrinsic helix stabilities and putative charge interactions in 434 Cro.