Reciprocal inhibition of human motoneurons with different firing frequencies

A study was made of the effect of reciprocal inhibition on individual firing motoneurons in the extensor carpi ulnaris and soleus muscle in human subjects. Peristimulus histograms (PSH) were plotted at different average frequency of motoneuron firing ( ) and the change in duration of interspike intervals (ISI) was analyzed. For reciprocal inhibition, as for other types of inhibition, is a factor in the effectiveness of motoneuron inhibition. The duration of inhibition apparent in the PSH, the sizes of zones of inhibition effectiveness in the ISIs and lengthening of the intervals are dependent on . For all motoneurons, the low range is most favorable for effective inhibition. The dependence of effectiveness of a volley on the time of its arrival within the ISI is also analyzed.Institute for Problems of Information Transmission, Russian Academy of Sciences, Moscow. Translated from Neirofiziologiya, Vol. 24, No. 6, pp. 643–653, November–December, 1992.     

Detection and analysis of recurrent inhibition of firing motoneurons in man

During regular firing of "small" motor units, activated during weak voluntary contraction of the human soleus muscle, thick efferent fibers of n. tibialis were stimulated (a small M response was evoked, in which the small units did not participate). Peristimulus histograms of potentials of single motor units were constructed and the effect of stimulation on interspike interval duration was analyzed. The firing rate of the motor units was 4.5–7.6 spikes/sec. Stimulation of the nerve led to a sharp decrease in probability of their discharge or even complete temporary cessation of firing, i.e., it had a well marked inhibitory effect (lasting 10–20 msec). The latent period of inhibition (35–40 msec) was only a little longer than the latent period of the monosynaptic reflex of the soleus muscle. The effect of an inhibitory volley on duration of the interspike interval of the motor units depended on the time when the volley arrived during the interval. Lengthening of the interval was observed only if the inhibitory volley arrived in the second half or at the end of the interval. It is concluded that inhibition of firing of small motor units is due to Renshaw cells, activated on stimulation of axons of large motoneurons. The efficiency of a short (compared with the duration of the interspike interval) inhibitory volley reaching a motoneuron firing at low frequency characteristic of its adequate activation, is discussed.Institute for Problems of Information Transmission, Academy of Sciences of the USSR, Moscow. Translated from Neirofiziologiya, Vol. 16, No. 1, pp. 88–96, January–February, 1984.     

Testing excitability in human firing motoneurons during the interspike interval

Monosynaptic testing of excitability in firing triceps surae muscle motoneurons activated during volitional contraction was performed using a technique for recording potentials from single motor units and by producing H-reflex. Motoneuronal excitability was assessed according to level of firing index. Motoneuronal firing index decreased during transition from a low background rhythmic firing rate of less than 6 spikes/sec to one of 6–8 spikes/sec. It hardly changed with a further rise in rate to 12 spikes/sec. The dependence between firing index and spike rate are put down to changes occurring in motoneuronal excitability during the interspike interval. Findings indicate that in the low frequency range of motoneuronal firing characteristic of natural muscle contraction, discharge rate may be considered one of the factors determining excitability in the motoneuron and hence its transmission qualities.Institute of Problems in Information Transmission, Academy of Sciences of the USSR. Translated from Neirofiziologiya, Vol. 19, No. 2, pp. 210–216, March–April, 1987.     

Reduction in maximal firing rate of motoneurons after 1-week immobilization of finger muscle in human subjects.

We investigated the effects of immobilization on the maximal motoneuronal firing rate recorded from the first dorsal interosseous (FDI) during voluntary isometric contraction. In five human subjects, the middle finger, index finger, and thumb were immobilized for 1 week in a fiber-glass cast, which kept FDI in a shortened position. During a maximal voluntary contraction, single muscle-fiber action potentials were recorded using a tungsten microelectrode, and mean firing rate was calculated for each action-potential train. Three recording sessions were held: before immobilization (pre), after immobilization (post), and after a 1-week recovery period (recovery). The mean firing rate of FDI motoneurons during maximal voluntary contraction was decreased immediately after the 1-week immobilization (pre: 39.0+/-3.2 Hz, number of detected spike trains (n)=353; post: 33.1+/-1.5 Hz, n=285; p<0.05), and there was a return to control after the recovery period (40.2+/-3.4 Hz, n=236). This suggests that the maximal motoneuronal firing rate achieved during maximal voluntary contraction is reduced after short-term immobilization. The functional implications and the contribution of this phenomenon to the immobilization-induced reduction in maximal voluntary force are discussed.     

Excitability of single firing human motoneurones to single and repetitive stimulation (experiment and model)

The activity of single motoneurones of m. flexor carpi ulnaris (FCU) was investigated by recording their motor unit (MU) action potentials during weak and moderate voluntary muscle contractions. The MU firing rate range was 4.5-15 imp/s. The excitability of motoneurones was tested with a number of single stimuli eliciting a monosynaptic H-reflex of low amplitude. Two different indices were defined which relate to motoneuronal excitability: the response index--the ratio of the number of responses of a motoneurone to the total number of stimuli, and the response time--the time after the last background MU discharge at which motoneurone is ready to respond to the excitatory volley. Both the response index and the response time were determined for single motoneurones at different levels of background activity. In the lower range of firing rates, the response index for all motoneurones decreased when increasing the firing rate, but it remained constant in the higher rate range. This kind of response seems to be a typical motoneuronal response to the stimulation with single stimuli. The data on the response time were used to study the excitability of the same single motoneurones to computer simulated repetitive stimulation (stimulation rate 40-100 imp/s). In this case, the excitability of each motoneurone was determined as an increment of its firing rate in response to the stimulation. For the lower firing rate range, the excitability for all motoneurones also decreased when the firing rates increased whereas a variety of slopes was obtained in the higher rate range.(ABSTRACT TRUNCATED AT 250 WORDS)     

The venous territories (venosomes) of the human body: experimental study and clinical implications

The venous architecture of the integument and the underlying deep tissues was studied in six total-body human fresh cadavers and a series of isolated regional studies of the limbs and torso. A radiopaque lead oxide mixture was injected, and the integument and deep tissues were dissected and radiographed. The sites of the venous perforators were plotted and traced to their underlying parent veins that accompany the source (segmental) arteries. A series of cross-sectional studies were made in one subject to illustrate the course of the perforators between the integument and the deep tissues. The veins were dissected under magnification to identify the site and orientation of the valves. Results revealed a large number of valveless (oscillating) veins within the integument and deep tissues that link adjacent valved venous territories and allow equilibration of flow and pressure throughout the tissue. Where choke arteries define the arterial territories, they are matched by boundaries of oscillating veins in the venous studies. The venous architecture is a continuous network of arcades that follow the connective-tissue framework of the body. The veins converge from mobile to fixed areas, and they "hitchhike" with nerves. The venous drainage mirrors the arterial supply in the deep tissues and in most areas of the integument in the head, neck, and torso. In the limbs, the stellate pattern of the venous perforators is modified by longitudinal channels in the subdermal network. However, when an island flap is raised, these longitudinal channels are disconnected, and once again the arterial and venous patterns match. Our venous studies add strength to the angiosome concept. Where source arteries supply a composite block of tissue, we have demonstrated radiologically and by microdissection that the branches of these arteries are accompanied by veins that drain in the opposite direction and return to the same locus. Hence each angiosome consists of matching arteriosomes and venosomes. The clinical implications of these results are discussed with particular reference to the design of flaps, the delay phenomenon, venous free flaps, the pathogenesis of flap necrosis, the "muscle pump," varicose veins, and venous ulceration.     

Selective inhibition of human acetylcholinesterase by xanthine derivatives: In vitro inhibition and molecular modeling investigations

The commonly used beverage and psychostimulant caffeine is known to inhibit human acetylcholinesterase enzyme. This pharmacological activity of caffeine is partly responsible for its cognition enhancing properties. However, the exact mechanisms of its binding to human cholinesterases (acetyl and butyrylcholinesterase; hAChE and hBuChE) are not well known. In this study, we investigated the cholinesterase inhibition by the xanthine derivatives caffeine, pentoxifylline, and propentofylline. Among them, propentofylline was the most potent AChE inhibitor (hAChE IC₅₀ = 6.40 μM). The hAChE inhibitory potency was of the order: caffeine (hAChE IC₅₀ = 7.25 μM) < pentoxifylline (hAChE IC₅₀ = 6.60 μM) ? propentofylline (hAChE IC₅₀ = 6.40 μM). These compounds were less potent relative to the reference agent donepezil (hAChE IC₅₀ = 0.04 μM). Moreover, they all exhibited selective inhibition of hAChE with no inhibition of hBuChE (IC₅₀ > 50 μM) relative to the reference agent donepezil (hBuChE IC₅₀ = 13.60 μM). Molecular modeling investigations indicate that caffeine binds primarily in the catalytic site (Ser203, Glu334 and His447) region of hAChE whereas pentoxifylline and propentofylline are able to bind to both the catalytic site and peripheral anionic site due to their increased bulk/size, thereby exhibiting superior AChE inhibition relative to caffeine. In contrast, their lack of hBuChE inhibition is due to a larger binding site and lack of key aromatic amino acids. In summary, our study has important implications in the development of novel caffeine derivatives as selective AChE inhibitors with potential application as cognitive enhancers and to treat various forms of dementia.     

Torsion of the human left ventricle: experimental analysis and computational modeling

We set a twofold investigation: we assess left ventricular (LV) rotation and twist in the human heart through 3D-echocardiographic speckle tracking, and use representative experimental data as benchmark with respect to numerical results obtained by solving our mechanical model of the LV. We aim at new insight into the relationships between myocardial contraction patterns and the overall behavior at the scale of the whole organ. It is concluded that torsional rotation is sensitive to transmural gradients of contractility which is assumed linearly related to action potential duration (APD). Pressure-volume loops and other basic strain measures are not affected by these gradients. Therefore, realistic torsional behavior of human LV may indeed correspond to the electrophysiological and functional differences between endocardial and epicardial cells recently observed in non-failing hearts. Future investigations need now to integrate the mechanical model proposed here with minimal models of human ventricular APD to drive excitation-contraction coupling transmurally.     

Leptin counteracts the hypoxia-induced inhibition of spontaneously firing hippocampal neurons: a microelectrode array study

Besides regulating energy balance and reducing body-weight, the adipokine leptin has been recently shown to be neuroprotective and antiapoptotic by promoting neuronal survival after excitotoxic and oxidative insults. Here, we investigated the firing properties of mouse hippocampal neurons and the effects of leptin pretreatment on hypoxic damage (2 hours, 3% O(2)). Experiments were carried out by means of the microelectrode array (MEA) technology, monitoring hippocampal neurons activity from 11 to 18 days in vitro (DIV). Under normoxic conditions, hippocampal neurons were spontaneously firing, either with prevailing isolated and randomly distributed spikes (11 DIV), or with patterns characterized by synchronized bursts (18 DIV). Exposure to hypoxia severely impaired the spontaneous activity of hippocampal neurons, reducing their firing frequency by 54% and 69%, at 11 and 18 DIV respectively, and synchronized their firing activity. Pretreatment with 50 nM leptin reduced the firing frequency of normoxic neurons and contrasted the hypoxia-induced depressive action, either by limiting the firing frequency reduction (at both ages) or by increasing it to 126% (in younger neurons). In order to find out whether leptin exerts its effect by activating large conductance Ca(2+)-activated K(+) channels (BK), as shown on rat hippocampal neurons, we applied the BK channel blocker paxilline (1 μM). Our data show that paxilline reversed the effects of leptin, both on normoxic and hypoxic neurons, suggesting that the adipokine counteracts hypoxia through BK channels activation in mouse hippocampal neurons.     

Ethanol utilization by sulfate-reducing bacteria: an experimental and modeling study

A mixed culture of sulfate-reducing bacteria containing the species Desulfovibrio desulfuricans was used to study sulfate-reduction stoichiometry and kinetics using ethanol as the carbon source. Growth yield was lower, and kinetics were slower, for ethanol compared to lactate. Ethanol was converted into acetate and no significant carbon dioxide production was observed. A mathematical model for growth of sulfate-reducing bacteria on ethanol was developed, and simulations of the growth experiments on ethanol were carried out using the model. The pH variation due to sulfate reduction, and hydrogen sulfide production and removal by nitrogen sparging, were examined. The modeling study is distinct from earlier models for systems using sulfate-reducing bacteria in that it considers growth on ethanol, and analyzes pH variations due to the product-formation reactions.     

A computational modeling and molecular dynamics study of the Michaelis complex of human protein Z-dependent protease inhibitor (ZPI) and factor Xa (FXa)

Protein Z-dependent protease inhibitor (ZPI) and antithrombin III (AT3) are members of the serpin superfamily of protease inhibitors that inhibit factor Xa (FXa) and other proteases in the coagulation pathway. While experimental structural information is available for the interaction of AT3 with FXa, at present there is no structural data regarding the interaction of ZPI with FXa, and the precise role of this interaction in the blood coagulation pathway is poorly understood. In an effort to gain a structural understanding of this system, we have built a solvent equilibrated three-dimensional structural model of the Michaelis complex of human ZPI/FXa using homology modeling, protein–protein docking and molecular dynamics simulation methods. Preliminary analysis of interactions at the complex interface from our simulations suggests that the interactions of the reactive center loop (RCL) and the exosite surface of ZPI with FXa are similar to those observed from X-ray crystal structure-based simulations of AT3/FXa. However, detailed comparison of our modeled structure of ZPI/FXa with that of AT3/FXa points to differences in interaction specificity at the reactive center and in the stability of the inhibitory complex, due to the presence of a tyrosine residue at the P1 position in ZPI, instead of the P1 arginine residue in AT3. The modeled structure also shows specific structural differences between AT3 and ZPI in the heparin-binding and flexible N-terminal tail regions. Our structural model of ZPI/FXa is also compatible with available experimental information regarding the importance for the inhibitory action of certain basic residues in FXa. Figure Solvent equilibrated models for protein z-dependent protease inhibitor and its initial reactive complex with coagulation factor Xa (show here) are developed. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users. V.C. and C.J.L. contributed equally to this work. The solvent-equilibrated PDB structure of the ZPI/FXa will be made available upon request. Conflict of interest statement  The authors state that they have no conflict of interest.     

Coerulospinal enhancement of repetitive firing with correlative changes in postspike afterhyperpolarization of cat spinal motoneurons

The present study was aimed at determining if inputs from the locus coeruleus (LC) have any effect on repetitive firing of ventral horn motoneurons in cats. In hindlimb flexor and extensor motoneurons stimulated intrasomatically with current below the threshold for repetitive discharges, added LC-evoked excitatory post-synaptic potentials (EPSPs) were consistently found to produce repetitive firing, suggesting a lowering in the repetitive firing threshold attributable to excitatory LC inputs. With those extensor motoneurons showing episodic, self-sustained firing, LC-EPSPs introduced during the quiescent period were capable of starting a continuous discharge with rhythmic frequencies higher than the spontaneous activity. In some of these cells, intracellularly applied hyperpolarizing current was able to stop the spontaneous discharges. Subsequently, LC stimuli were found to reinitiate repetitive discharges, thus counteracting the ongoing suppression of the motoneurons. Quantitative analysis of the single-spike afterhyperpolarization (AHP) indicated a consistent reduction in its amplitude and time course (duration, time-to-peak, half-decay time) for flexor and extensor motoneurons in response to LC conditioning stimuli. Present results suggest an excitatory LC action on the repetitive discharges of cat motoneurons accompanied by a concurrent decrease in the amplitude and time course of the single-spike AHPs.     

Ribonucleotide reductase inhibition by metal complexes of Triapine (3-aminopyridine-2-carboxaldehyde thiosemicarbazone): a combined experimental and theoretical study

Triapine (3-aminopyridine-2-carboxaldehyde thiosemicarbazone, 3-AP) is currently the most promising chemotherapeutic compound among the class of α-N-heterocyclic thiosemicarbazones. Here we report further insights into the mechanism(s) of anticancer drug activity and inhibition of mouse ribonucleotide reductase (RNR) by Triapine. In addition to the metal-free ligand, its iron(III), gallium(III), zinc(II) and copper(II) complexes were studied, aiming to correlate their cytotoxic activities with their effects on the diferric/tyrosyl radical center of the RNR enzyme in vitro. In this study we propose for the first time a potential specific binding pocket for Triapine on the surface of the mouse R2 RNR protein. In our mechanistic model, interaction with Triapine results in the labilization of the diferric center in the R2 protein. Subsequently the Triapine molecules act as iron chelators. In the absence of external reductants, and in presence of the mouse R2 RNR protein, catalytic amounts of the iron(III)–Triapine are reduced to the iron(II)–Triapine complex. In the presence of an external reductant (dithiothreitol), stoichiometric amounts of the potently reactive iron(II)–Triapine complex are formed. Formation of the iron(II)–Triapine complex, as the essential part of the reaction outcome, promotes further reactions with molecular oxygen, which give rise to reactive oxygen species (ROS) and thereby damage the RNR enzyme. Triapine affects the diferric center of the mouse R2 protein and, unlike hydroxyurea, is not a potent reductant, not likely to act directly on the tyrosyl radical.