Modified classification of normal lung sounds applying Quantile vectors.

In this paper a novel Lung Sound Automatic Verification (LSAV) system and front-end Quantile based acoustic models to classify Lung Sounds (LS) are proposed. The utilization of Quantiles allowed an easier and objective assessment with smaller computational demand. Moreover, less-complex Gaussian Mixture Models (GMM) were computed than those previously reported. The LSAV system allowed us to reach practically negligible error in healthy (normal) LS verification. LASV system efficiency and the optimal GMM's were evaluated by using Equal Error Rate (EER) and Bayesian Information Criterion (BIC) techniques respectively. These approaches could provide a tool for broader medical evaluation which does not rely, as it is often the case, on a qualitative and subjective description of LS.

Tailoring magnetic field gradient design to magnet cryostat geometry.

Eddy currents induced within a magnetic resonance imaging (MRI) cryostat bore during pulsing of gradient coils can be applied constructively together with the gradient currents that generate them, to obtain good quality gradient uniformities within a specified imaging volume over time. This can be achieved by simultaneously optimizing the spatial distribution and temporal pre-emphasis of the gradient coil current, to account for the spatial and temporal variation of the secondary magnetic fields due to the induced eddy currents. This method allows the tailored design of gradient coil/magnet configurations and consequent engineering trade-offs. To compute the transient eddy currents within a realistic cryostat vessel, a low-frequency finite-difference time-domain (FDTD) method using total-field scattered-field (TFSF) scheme has been performed and validated.

A sensory neuron subpopulation with unique sequential survival dependence on nerve growth factor and basic fibroblast growth factor during development.

We characterized a subpopulation of dorsal root ganglion (DRG) sensory neurons that were previously identified as preferential targets of enkephalins. This group, termed P-neurons after their "pear" shape, sequentially required nerve growth factor (NGF) and basic fibroblast growth factor (bFGF) for survival in vitro during different developmental stages. Embryonic P-neurons required NGF, but not bFGF. NGF continued to promote their survival, although less potently, up to postnatal day 2 (P2). Conversely, at P5, they needed bFGF but not NGF, with either factor having similar effects at P2. This trophic switch was unique to that DRG neuronal group. In addition, neither neurotrophin-3 (NT-3) nor brain-derived neurotrophic factor influenced their survival during embryonic and postnatal stages, respectively. The expression of NGF (Trk-A) and bFGF (flg) receptors paralleled the switch in trophic requirement. No single P-neuron appeared to coexpress both Trk-A and flg. In contrast, all of them coexpressed flg and substance P, providing a specific marker of these cells. Immunosuppression of bFGF in newborn animals greatly reduced their number, suggesting that the factor was required in vivo. bFGF was present in the DRG and spinal cord, as well as in skeletal muscle, the peripheral projection site of P-neurons, as revealed by tracer DiIC(18)3. The lack of requirement of NT-3 for survival and immunoreactivity for the neurofilament of 200 kDa distinguished them from muscle proprioceptors, suggesting that they are likely to be unmyelinated muscle fibers. Collectively, their properties indicate that P-neurons constitute a distinct subpopulation of sensory neurons for which the function may be modulated by enkephalins.

delta opioid receptor modulation of several voltage-dependent Ca(2+) currents in rat sensory neurons.

Endogenous enkephalins and delta opiates affect sensory function and pain sensation by inhibiting synaptic transmission in sensory circuits via delta opioid receptors (DORs). DORs have long been suspected of mediating these effects by modulating voltage-dependent Ca(2+) entry in primary sensory neurons. However, not only has this hypothesis never been validated in these cells, but in fact several previous studies have only turned up negative results. By using whole-cell current recordings, we show that the delta enkephalin analog [D-Ala(2), D-Leu(5)]-enkephalin (DADLE) inhibits, via DORs, L-, N-, P-, and Q-high voltage-activated Ca(2+) channel currents in cultured rat dorsal root ganglion (DRG) neurons. The percentage of responding cells was remarkably high (75%) within a novel subpopulation of substance P-containing neurons compared with the other cells (18-35%). DADLE (1 microM) inhibited 32% of the total barium current through calcium channels (I(Ba)). A delta (naltrindole, 1 microM), but not a mu (beta-funaltrexamine, 5 microM), antagonist prevented the DADLE response, whereas a DOR-2 subtype (deltorphin-II, 100 nM), but not a DOR-1 (DPDPE, 1 microM), agonist mimicked the response. L-, N-, P-, and Q-type currents contributed, on average, 18, 48, 14, and 16% to the total I(Ba) and 19, 50, 26, and 20% to the DADLE-sensitive current, respectively. The drug-insensitive R-type current component was not affected by the agonist. This work represents the first demonstration that DORs modulate Ca(2+) entry in sensory neurons and suggests that delta opioids could affect diverse Ca(2+)-dependent processes linked to Ca(2+) influx through different high-voltage-activated channel types.

Stable expression and regulation of a rat brain K+ channel.

The Shaw-type K+ channel Kv3.1 was stably transfected in human embryonic kidney cells. Voltage dependence of activation, K+ permeability, sensitivity to external tetraethylammonium, and unitary conductance were similar to Kv3.1 channels expressed transiently in Xenopus oocytes. Kv3.1 channels appear to be regulated because the protein kinase C activator phorbol 12,13-dibutyrate decreased Kv3.1 currents. Based on these results, we find that the stable expression of voltage-gated K+ channels in human embryonic kidney cells appears to be well suited for analysis of both biophysical and biochemical regulatory processes.

Neuromodulation.

The ability of the nervous system to respond to the environment and to learn depends upon the tuning of neuronal electrical activity, loosely called neuromodulation. The substrates for electrical activity and, therefore, neuromodulation are ion channels which may be either synaptic or extrasynaptic. Neuromodulation is dynamic and most frequently involves neurotransmitters and hormones acting via G-protein-coupled pathways.

Kinetics of G protein-mediated modulation of the potassium M-current in bullfrog sympathetic neurons.

The inhibition of the voltage-dependent, K+ M-current (IM) following receptor-independent G protein activation with controlled intracellular perfusion of nonhydrolyzable GTP analogs had an exponential time course, with rates hyperbolically dependent on GTP analog concentration, and a limiting value of 0.53 min-1. The inhibitory agonist muscarine caused a concentration-dependent acceleration of the rate of nucleotide-induced inhibition, with a plateau of about 20 min-1 and an exponential time course. In neurons not treated with nucleotide analogs the IM recovery rate following agonist removal was 3-7 min-1. It is proposed that the overall kinetics of the transduction pathway for IM modulation is governed by the agonist-dependent kinetics of nucleotide interaction with G proteins. A simple model of IM modulation based on G proteins' kinetics has been developed. These data suggest a possible cellular process responsible for the time course of slow synaptic potentials caused by IM inhibition in sympathetic neurons.

Correlation between G protein activation and reblocking kinetics of Ca2+ channel currents in rat sensory neurons.

Membrane depolarization relieves the G protein-mediated inhibition or block of high threshold Ca2+ channel currents. We found that the net rate of reblocking depended on the extent of G protein activation. With low intracellular concentrations of GTP gamma S reblocking rates resembled inactivation rates; with higher concentrations reblocking rates increased progressively. Reblocking kinetics were fit with a sum of two exponential functions having time constants (in ms) tau F greater than or equal to 10 and tau S greater than or equal to 30. Unblock during depolarization was fit by a single exponential function with time constant tau A similar to tau F. A model was developed in which unblocking followed dissociation of a blocking molecule, possibly the G protein itself, from Ca2+ channels, and reblocking occurred at rates that depended on the concentration of the blocking molecule. The time course of Ca2+ entry and thus presynaptic Ca2+ levels can be regulated by both the concentration of the G-protein-dependent blocking particle and membrane potential.

Associative synaptic potentiation and depression: quantification of dissociable modifications in the hippocampal dentate gyrus favors a particular class of synaptic modification equations.

This report further characterizes associative long-term synaptic modification of the ipsilateral and contralateral synapses formed by the bilateral entorhinal cortical (EC) projection to the dentate gyrus (DG). The experimental model is the anesthetized hooded rat. The quantitative results qualify this system as a model for studying the rules of associative synaptic modification formulated in terms of individual synapses. Bilateral DG microelectrodes recorded both ipsilateral and contralateral EC-DG responses before and after brief, high-frequency EC conditioning stimulation. The weak contralateral pathway received high-frequency conditioning before, during, or after similar conditioning of the strong, converging ipsilateral pathway. Statistical analyses revealed two types of significant, dissociated synaptic modifications, which depend on the relationship of the ipsilateral and contralateral afferents. First, contralateral EC-DG responses potentiated or depressed when the converging ipsilateral responses concurrently either potentiated or remained unchanged. Second, contralateral EC-DG responses potentiated, depressed, or showed no change when the collateral ipsilateral responses concurrently either potentiated or remained unchanged. Correlation and contingency table analyses indicated that changes in the contralateral synaptic responses are not well predicted by changes at either neighboring synapses of the converging ipsilateral pathway or at synapses of the collateral ipsilateral pathway. The contingencies of associated pre- and postsynaptic activation determined by the conditioning paradigm, however, accurately predicted the altered synaptic responses of both ipsilateral and contralateral EC-DG pathways. The results imply that associative synaptic modification in the EC-DG system is specific to individual synapses and requires both appropriate presynaptic and postsynaptic activation. Because this system provides suitable controls for nonspecific effects of conditioning stimulation and because modification of neighboring synapses is dissociable, the EC-DG system can be used to study further those rules of activity-dependent associative modification that are formulated in terms of individual synapses. The discussion briefly considers published rules of synaptic modification, pointing out several rules that are not consistent with the experimental observations and one that agrees with the present results.

A G Protein Mediates the Inhibition of the Voltage-Dependent Potassium M Current by Muscarine, LHRH, Substance P and UTP in Bullfrog Sympathetic Neurons.

The involvement of G proteins in the transduction mechanism of M current (Im) inhibition by extracellular ligands in bullfrog sympathetic neurons was examined using the hydrolysis resistant nucleotide analogues GTPgammaS and GDPbetaS. Im was recorded in large (40 - 60 microm) isolated neurons using the patch-clamp technique in the whole-cell configuration, as well as in neurons from the intact ganglion impaled with conventional microelectrodes. In whole-cell recordings Im could be recorded without significant loss for 1 h or more provided ATP was present in the patch pipette. Muscarine, D-Ala6-LHRH, substance P and UTP reversibly inhibited Im in isolated control neurons, with full and rapid recovery of the current following agonist washout. Dialysis of isolated neurons with various concentrations of GTPgammaS (1 - 100 microM) affected, in a dose-dependent manner, the recovery of Im after its inhibition by brief agonist application. With 50 microM GTPgammaS, Im inhibition became completely irreversible. Similarly, the reversibility of Im inhibition by muscarine was reduced or abolished by the iontophoretic injection of GTPgammaS through a second microelectrode into neurons of the intact ganglion. GTPgammaS by itself caused a slow, agonist-independent suppression of Im in dialysed neurons, thus mimicking agonist action. Dialysis of isolated neurons with GDPbetaS (100 - 500 microM) attenuated by half or more the magnitude of Im inhibition by agonist as compared to control neurons. In addition, GDPbetaS attenuated the response of a given neuron to muscarine and D-Ala6-LHRH, and caused slow increase of Im, as a function of dialysis time. Incubation (2 - 72 h, 4 - 36 degrees C) of isolated neurons or intact ganglions with activated pertussis toxin had no effect on the response to muscarine. Toxin injections to experimental animals were equally ineffective. In contrast to Im, the additional inward current with increase in conductance induced by muscarine and D-Ala6-LHRH reversed with agonist washout in GTPgammaS-dialysed neurons, although more slowly than in control neurons. The results in this study indicate that a G protein, possibly pertussis toxin-insensitive, provides a common coupling step linking muscarinic, substance P, D-Ala6-LHRH and UTP receptors to the inhibition of M current.

Evidence for peripeduncular neurons having a role in the control of feminine sexual behavior in the rat.

In order to test the hypothesis that peripeduncular nucleus (PPN) cells are essential for normal sexual behavior, synaptic blockade in the chronically implanted awake animal was attempted by means of the local injection of pentobarbital (PB) at a concentration which may interfere with synaptic transmission without affecting conduction in fibers. Ovariectomized, female rats were chronically implanted with cannulae directed to the PPN or the dorsal midbrain. After adequate estrogen priming, rats were injected with 22 mM pentobarbital (PB) or artificial cerebrospinal fluid (ACSF). Seven min after PB injection in PPN lordosis quotient was significantly lowered, whereas PB in dorsal midbrain or ACSF in PPN had no effect. It is concluded that PPN neurons themselves may have a functional role in the control of lordosis and that loss of PPN neurons may account for impairment of reproductive behavior observed after lesions in the ventrolateral midbrain.

Further studies on peripeduncular-hypothalamic pathways involved in sexual behavior in the female rat.

A series of preliminary experiments demonstrated that injection of 22 mM sodium pentobarbital in the brain of the rat blocked synaptic transmission at the site of injection; the same concentration of pentobarbital did not block fiber conduction. Based on the latter information, 22 mM pentobarbital was applied to different parts of the peripeduncular-hypothalamic pathways responsible for the conduction and generation of potentials evoked in the ventromedial nucleus (VMN) by stimuli applied to the peripeduncular nucleus (PPN), to determine whether participation of the amygdala and bed nucleus of the stria terminalis involves the transynaptic activation of neuron somas at these places or the operation of passing fibers only. We determined that potentials evoked in the VMN by PPN stimulation involves synaptic activity in both the lateral amygdaloid nucleus and the bed nucleus of the stria terminalis. Both structures receive PPN-originated activity independently, and both structures contribute to the generation of PPN-VMN evoked responses, presumably through temporal or spatial summation of inputs in the VMN. We also showed that activity in the lateral amygdaloid nucleus is conducted toward the VMN along fibers in the stria terminalis. We propose that the synaptic interactions thus demonstrated serve as integrating relays for different sensory modalities and hormone actions regulating sexual behavior.

Investigation of peripeduncular-hypothalamic pathways involved in the control of lordosis in the rat.

Single shock stimuli applied to the peripeduncular nucleus (PPN) elicited a complex evoked response in the hypothalamic ventromedial nucleus (VMN). An early component and a late component could be distinguished in the evoked response on the basis of their different latency, threshold, site of maximal amplitude, frequency-response characteristics and also because restricted lesions eliminated specifically the short-or the long-latency components. Transection of the dorsal supraoptic pathway immediately in front of the PPN suppressed all the VMN-evoked responses. The same lesion eliminated lordotic responses in ovariectomized rats treated with estradiol benzoate and progesterone. Similar lesions placed more dorsally had no effect on either the sexual behavior or the evoked response. Evidence was obtained suggesting that the short-latency component is generated by activity that reaches the VMN directly through the ventral supraoptic commissure, while the long-latency response involves substations in the amygdala and the bed nucleus of the stria terminalis. The effect of lesions on the performance of lordosis may be attributed to the disruption of ascending and/or descending neural impulses circulating between the PPN and the VMN, with relay stations in the amygdala and the bed nucleus of the stria terminalis.