Biomedical subjects

D Contreras

Publications and source records attributed to D Contreras.

At least 19 recordsLinked to original sources

PubMed2005

Comparison of responses to electrical stimulation and whisker deflection using two different voltage-sensitive dyes in mouse barrel cortex in vivo.

We examined the spatial structure of noise in optical recordings made with two commonly used voltage-sensitive dyes (RH795 and RH1691) in mouse barrel cortex in vivo, and determined that the signal-to-noise ratio of the two dyes was comparable when averaging over barrel-sized areas, or at single pixels distant from large blood vessels. We examined the spatiotemporal development of whisker- and electrically-evoked optical responses by quantifying the area of activated cortical surface as a function of time. Whisker and electrical stimuli activated cortical areas between 0.2-2.0 mm(2) depending on intensity. More importantly, both types of activation recruited cortical area at similar rates and showed a linear relationship between the maximal activated area and the peak rate of increase of the activated area. We propose a general rule of supragranular cortical activation in which the initial spreading speed of the response determines the total activated area, independent of the type of activation. Finally, despite comparable single-response kinetics, we observed greater paired-pulse depression of whisker-evoked responses relative to electrically-evoked responses.

Animals↗

PubMed2004

Fe(III)-EDTA complex abatement using a catechol driven Fenton reaction combined with a biological treatment.

A combined chemical oxidation (catechol-driven Fenton reaction) followed by a biological treatment was used to degrade Fe(III)-EDTA (1.34 mM). The chemical treatment was inspired in fungal non-enzymatic wood rot mechanisms that use dihydroxybenzens in order to promote the Fenton reaction to breakdown wood structures. This chemical pre-oxidation used catechol (50 microM) and H202 (20 mM) and the reaction products were identified by GC-MS. In addition, a biological treatment was coupled using the baker's yeast Saccharomyces cerevisiae. The combined chemical biological treatment achieved 100% EDTA degradation, 68 % total organic carbon removal and 90% iron removal.

Biodegradation, Environmental↗

PubMed2004

Degradation of recalcitrant compounds by catechol-driven Fenton reaction.

Dihydroxybenzenes are able to reduce Fe(III) and promote the Fenton reaction in the presence of H2O2. The catechol/Fe(III)/H2O2 system has been successfully used to degrade different compounds, being more efficient than the Fe(II)-Fenton reaction. In this paper the possibilities for using the catechol-driven Fenton reaction to degrade recalcitrant compounds such as the Fe(III)-EDTA complex and veratryl alcohol are reviewed.

Benzene Derivatives↗

PubMed2001

Voltage-sensitive dye imaging of neocortical spatiotemporal dynamics to afferent activation frequency.

The spatial and temporal patterns of neocortex activation are determined not only by the dynamic character of the input but also by the intrinsic dynamics of the cortical circuitry. To study the role of afferent input frequency on cortical activation dynamics, the electrical activity of in vitro neocortex slices was imaged during white-matter electrical stimulation. High-speed optical imaging was implemented using voltage-sensitive dyes in guinea pig visual and somatosensory cortex slices concomitantly with intracellular recordings. Single white-matter electrical stimuli activated well-defined cortical sites with a radially oriented columnar configuration. This configuration was followed, over the next few milliseconds, by a lateral spread of excitation through cortical layers 5 and 6 and layers 2 and 3. Much of the optical response was eliminated in low extracellular calcium, indicating that it was primarily synaptically mediated. Repetitive stimuli at 10 Hz reproduced the spatiotemporal pattern observed for single stimuli. In contrast, repetitive stimulation in the gamma frequency range ( approximately 40 Hz) rapidly restrained the area of excitation to a small columnar site directly above the stimulating electrode. Intracellular recordings from cells lateral to the activated column revealed increased inhibitory synaptic activity and/or decreased excitatory responses during the train at 40 Hz, but not during a 10 Hz stimulation. Localized microinjections of GABA(A) antagonist produced a reorganization of the geometrical activity pattern that was dependent on the position of the microinjection site. These findings indicate that the frequency-dependent spatial organization of neocortex activation is determined by inhibitory sculpting attributable to local network dynamics.

Afferent Pathways↗

PubMed2001

On the cellular and network bases of epileptic seizures.

The highly interconnected networks of the mammalian forebrain can generate a wide variety of synchronized activities, including those underlying epileptic seizures, which often appear as a transformation of otherwise normal brain rhythms. The cerebral cortex and hippocampus are particularly prone to the generation of the large, synchronized bursts of activity underlying many forms of seizures owing to strong recurrent excitatory connections, the presence of intrinsically burst-generating neurons, ephaptic interactions among closely spaced neurons, and synaptic plasticity. The simplest form of epileptiform activity in these structures is the interictal spike, a synchronized burst of action potentials generated by recurrent excitation, followed by a period of hyperpolarization, in a localized pool of pyramidal neurons. Seizures can also be generated in response to a loss of balance between excitatory and inhibitory influences and can take the form of either tonic depolarizations or repetitive, rhythmic burst discharges, either as clonic or spike-wave activity, again mediated both by intrinsic membrane properties and synaptic interactions. The interaction of the cerebral cortex and the thalamus, in conjunction with intrathalamic communication, can also generate spike waves similar to those occurring during human absence seizure discharges. Although epileptic syndromes and their causes are diverse, the cellular mechanisms of seizure generation appear to fall into only two categories: rhythmic or tonic "runaway" excitation or the synchronized and rhythmic interplay between excitatory and inhibitory neurons and membrane conductances.

Animals↗

PubMed2000

Impaired fast-spiking, suppressed cortical inhibition, and increased susceptibility to seizures in mice lacking Kv3.2 K+ channel proteins.

Voltage-gated K(+) channels of the Kv3 subfamily have unusual electrophysiological properties, including activation at very depolarized voltages (positive to -10 mV) and very fast deactivation rates, suggesting special roles in neuronal excitability. In the brain, Kv3 channels are prominently expressed in select neuronal populations, which include fast-spiking (FS) GABAergic interneurons of the neocortex, hippocampus, and caudate, as well as other high-frequency firing neurons. Although evidence points to a key role in high-frequency firing, a definitive understanding of the function of these channels has been hampered by a lack of selective pharmacological tools. We therefore generated mouse lines in which one of the Kv3 genes, Kv3.2, was disrupted by gene-targeting methods. Whole-cell electrophysiological recording showed that the ability to fire spikes at high frequencies was impaired in immunocytochemically identified FS interneurons of deep cortical layers (5-6) in which Kv3.2 proteins are normally prominent. No such impairment was found for FS neurons of superficial layers (2-4) in which Kv3.2 proteins are normally only weakly expressed. These data directly support the hypothesis that Kv3 channels are necessary for high-frequency firing. Moreover, we found that Kv3.2 -/- mice showed specific alterations in their cortical EEG patterns and an increased susceptibility to epileptic seizures consistent with an impairment of cortical inhibitory mechanisms. This implies that, rather than producing hyperexcitability of the inhibitory interneurons, Kv3.2 channel elimination suppresses their activity. These data suggest that normal cortical operations depend on the ability of inhibitory interneurons to generate high-frequency firing.

Action Potentials↗

PubMed2000

The obstetrician-gynecologist's role in vaccine-preventable diseases and immunization.

OBJECTIVE: To assess by survey the immunization role currently played obstetrician-gynecologists in the state of Michigan. METHODS: Masked questionnaires requesting demographic, knowledge-based, practice, and attitudinal data were sent to 850 ACOG-registered fellows. RESULTS: Three hundred sixty-five physicians responded, 313 of whom were in active practice. Most were male (70%) and graduated from medical school between 1970 and 1989 (68%). The majority provided both obstetric and gynecologic services. The minority (47%) specifically identified themselves as primary care providers. Only 15% of respondents considered screening for vaccine-preventable diseases to be outside the realm of routine obstetric-gynecologic care. In practice, however, 19% did not screen their obstetric patients for any vaccine-preventable diseases, and only 10% assessed their patients for all nine vaccine-preventable diseases listed in the questionnaire. In gynecologic patients, almost 40% of physicians did not assess for any vaccine-preventable disease. A wide range in knowledge level was identified concerning vaccine-preventable diseases, immunization recommendations, and vaccine safety. CONCLUSION: These data show a discrepancy between perceived responsibilities and actual practice patterns of obstetrician-gynecologists regarding vaccine-preventable diseases and the immunization of women. Limitations in current knowledge and practical concerns specific to vaccine administration contribute to this disparity.

Female↗

PubMed2000

Intralesional recombinant interferon alpha-2b in Peyronie's disease.

OBJECTIVE: To evaluate interferon alpha-2b (IFN) in the treatment of Peyronie's disease (PD) since IFN exerts antifibrotic action through collagen synthesis inhibition and fibrolysis stimulation. METHODS: The study comprised 34 patients, aged 31 to 63, with clinical and ultrasonographic (US) diagnosis of PD, who gave their consent to enter the study. They had the disease for 10.1 +/- 5.6 (2-22) months. Ten million IU of IFN were injected intralesionally, twice weekly for 14 weeks or less if there was complete remission. Clinical evaluation included penis angle at erection, sexual dysfunction (pain, possibility of intercourse) and palpable plaque. Plaque size was evaluated by US. Systemic and local adverse reactions, and anti-IFN antibodies were monitored as well. RESULTS: Sexual dysfunction disappeared in 19/24 (79.2%) patients with this disorder, palpable lesions in 21/34 (62%), angle at erection in 15/32 (47%), and pain in 16/17 (94%). Complete clinical response was achieved in 16/34 patients (47%). Ultrasonographic response rate was 88%, (53% complete). Plaque size decreased from 56.7 +/- 42.9 (median: 35.4) before treatment to 12.7 +/- 22.6 mm2 (median: 0) (p < 0.00001; Wilcoxon's paired test). Clinical and US responses correlated. No patient showed progression. Eight of 9 patients in whom other treatments had failed responded to IFN therapy (5 complete). The main systemic adverse reaction in most patients (mild or moderate) was the flu-like syndrome expected for IFN. Local reactions, more related to the administration procedure than to IFN itself, were small hematoma (10 patients), edema (3), cysts that were excised surgically (2), and venous leak (1). No patient developed anti-IFN antibodies. CONCLUSIONS: IFN treatment can be a suitable option for the management of PD. The results appear to be better than those achieved with other procedures. Further work should include comparative studies, long-term follow-up of treated patients, and alternative ways of administration.

Adult↗

PubMed1999

Spatiotemporal analysis of local field potentials and unit discharges in cat cerebral cortex during natural wake and sleep states.

The electroencephalogram displays various oscillation patterns during wake and sleep states, but their spatiotemporal distribution is not completely known. Local field potentials (LFPs) and multiunits were recorded simultaneously in the cerebral cortex (areas 5-7) of naturally sleeping and awake cats. Slow-wave sleep (SWS) was characterized by oscillations in the slow (<1 Hz) and delta (1-4 Hz) frequency range. The high-amplitude slow-wave complexes consisted in a positivity of depth LFP, associated with neuronal silence, followed by a sharp LFP negativity, correlated with an increase of firing. This pattern was of remarkable spatiotemporal coherence, because silences and increased firing occurred simultaneously in units recorded within a 7 mm distance in the cortex. During wake and rapid-eye-movement (REM) sleep, single units fired tonically, whereas LFPs displayed low-amplitude fast activities with increased power in fast frequencies (15-75 Hz). In contrast with the widespread synchronization during SWS, fast oscillations during REM and wake periods were synchronized only within neighboring electrodes and small time windows (100-500 msec). This local synchrony occurred in an apparent irregular manner, both spatially and temporally. Brief periods (<1 sec) of fast oscillations were also present during SWS in between slow-wave complexes. During these brief periods, the spatial and temporal coherence, as well as the relation between units and LFPs, was identical to that of fast oscillations of wake or REM sleep. These results show that natural SWS in cats is characterized by slow-wave complexes, synchronized over large cortical territories, interleaved with brief periods of fast oscillations, characterized by local synchrony, and of characteristics similar to that of the sustained fast oscillations of activated states.

Animals↗

PubMed1999

Cortically-induced coherence of a thalamic-generated oscillation.

Oscillatory patterns in neocortical electrical activity show various degrees of large-scale synchrony depending on experimental conditions, but the exact mechanisms underlying these variations of coherence are not known. Analysis of multisite local field potentials revealed that the coherence of spindle oscillations varied during different states. During natural sleep, the coherence was remarkably high over cortical distances of several millimeters, but could be disrupted by artificial cortical depression, similar to the effect of barbiturates. Possible mechanisms for these variations of coherence were investigated by computational models of interacting cortical and thalamic neurons, including their intrinsic firing patterns and various synaptic receptors present in the circuitry. The model indicates that modulation of the excitability of the cortex can affect spatiotemporal coherence with no change in the thalamus. The highest level of coherence was obtained by enhancing the excitability of cortical pyramidal cells, simulating the action of neuromodulators such as acetylcholine and noradrenaline. The underlying mechanism was due to cortex-thalamus-cortex loops in which a more excitable cortical network generated a more powerful and coherent feedback onto the thalamus, resulting in highly coherent oscillations, similar to the properties measured during natural sleep. In conclusion, these experiments and models are compatible with a powerful role for the cortex in triggering and synchronizing oscillations generated in the thalamus, through corticothalamic feedback projections. The model suggests that intracortical mechanisms may be responsible for synchronizing oscillations over cortical distances of several millimeters through cortex-thalamus-cortex loops, thus providing a possible cellular mechanism to explain the genesis of large-scale coherent oscillations in the thalamocortical system.

Anesthesia↗

PubMed1999

Pathomorphometrical characteristics of atherosclerosis in youth. A multinational investigation of WHO/World Heart Federation (1986-1996), using atherometric system.

BACKGROUND AND AIM: From 1986 to 1996, 1339 autopsies were performed on children and young adults, aged 5-34 years, in 18 countries of five continents in the course of the multinational investigation of the World Health Organization/International Society and Federation Cardiology (WHO/ISFC), "Pathobiological Determinants of Atherosclerosis in Youth" (PBDAY). A set of 966 left-half thoracic and 947 left-half abdominal aortae and 958 right coronary arteries were processed in the Center of Investigations and References of Atherosclerosis of Havana (CIRAH), i.e., one of the Reference Centers of the PBDAY. Pathomorphological and morphometrical analyses were carried out by a well-established method, the Atherometric System (AS). METHODS AND RESULTS: By qualitative analysis AS permitted the identification of each type of atherosclerotic lesions (AL). The quantitative analysis, using a digitizer (MYPAC-Japan, a PC-Pentium 200 Mhz-32 MB RAM), and the software Atherosoft, allowed the measurement of the intima surface occupied by any kind of AL, and estimation of the volume occupied and thus the degree of obstruction and stenosis of the lumen. The autopsy data were divided into three age groups: a) 5 to 14 years; b) 15 to 24 years and c) 25 to 34 and processed by age and sex. The commercial package NCSS was utilized for statistical analysis of the data. CONCLUSIONS: Of particular interest were the following findings: a) Atherosclerosis increases with age; b) Fatty streaks (FS) were always present already at 5 years of age, independent of the country, climate, state of nourishment, type and amount of foods and the habits and lifestyle of the population studied. FS progressed most rapidly from 15 to 24 years. The fibrous plaque began to appear slowly at the end of the second and rapidly progressed after the third decades. The severe plaque was rarely observed before 30 years of age; it appeared in the fourth decade and then progressed slowly, but steadily.

Adolescent↗

PubMed1998

The neuronal basis for consciousness.

Attempting to understand how the brain, as a whole, might be organized seems, for the first time, to be a serious topic of inquiry. One aspect of its neuronal organization that seems particularly central to global function is the rich thalamocortical interconnectivity, and most particularly the reciprocal nature of the thalamocortical neuronal loop function. Moreover, the interaction between the specific and non-specific thalamic loops suggests that rather than a gate into the brain, the thalamus represents a hub from which any site in the cortex can communicate with any other such site or sites. The goal of this paper is to explore the basic assumption that large-scale, temporal coincidence of specific and non-specific thalamic activity generates the functional states that characterize human cognition.

Animals↗

PubMed1998

Mechanisms underlying the synchronizing action of corticothalamic feedback through inhibition of thalamic relay cells.

Early studies have shown that spindle oscillations are generated in the thalamus and are synchronized over wide cortical territories. More recent experiments have shown that this large-scale synchrony depends on the integrity of corticothalamic feedback. Previously proposed mechanisms emphasized exclusively intrathalamic mechanisms to generate the synchrony of these oscillations. In the present paper, we propose a cellular mechanism in which the synchrony is dependent of a mutual interaction between cortex and thalamus. This cellular mechanism is tested by computational models consisting of pyramidal cells, interneurons, thalamic reticular (RE) and thalamocortical (TC) relay cells, on the basis of voltage-clamp data on intrinsic currents and synaptic receptors present in the circuitry. The model suggests that corticothalamic feedback must operate on the thalamus mainly through excitation of GABAergic RE neurons, therefore recruiting relay cells essentially through inhibition and rebound. We provide experimental evidence for such dominant inhibition in the lateral posterior nucleus. In these conditions, the model shows that cortical discharges optimally evoked thalamic oscillations. This feature is essential to the present cellular mechanism and is also consistently observed experimentally. The model further shows that, with this type of corticothalamic feedback, cortical discharges recruited large areas of the thalamus because of the divergent cortex-to-RE and RE-to-TC axonal projections. Consequently, the thalamocortical network generated patterns of oscillations and synchrony similar to in vivo recordings. The model also emphasizes the important role of the modulation of the Ih current by calcium in TC cells. This property conferred a relative refractoriness to the entire network, a feature also observed experimentally, as we show here. Further, the same property accounted for various spatiotemporal features of oscillations, such as systematic propagation after low-intensity cortical stimulation, local oscillations, and more generally, a high variability in the patterns of spontaneous oscillations, similar to in vivo recordings. We propose that the large-scale synchrony of spindle oscillations in vivo is the result of thalamocortical interactions in which the corticothalamic feedback acts predominantly through the RE nucleus. Several predictions are suggested to test the validity of this model.

Animals↗

PubMed1998

Spike-wave complexes and fast components of cortically generated seizures. I. Role of neocortex and thalamus.

We explored the relative contributions of cortical and thalamic neuronal networks in the generation of electrical seizures that include spike-wave (SW) and polyspike-wave (PSW) complexes. Seizures were induced by systemic or local cortical injections of bicuculline, a gamma-aminobutyric acid-A (GABAA) antagonist, in cats under barbiturate anesthesia. Field potentials and extracellular neuronal discharges were recorded through arrays of eight tungsten electrodes (0.4 or 1 mm apart) placed over the cortical suprasylvian gyrus and within the thalamus. 1) Systemic injections of bicuculline induced SW/PSW seizures in cortex, whereas spindle sequences continued to be present in the thalamus. 2) Cortical suprasylvian injection of bicuculline induced focal paroxysmal single spikes that developed into full-blown seizures throughout the suprasylvian cortex. The seizures were characterized by highly synchronized SW or PSW complexes at 2-4 Hz, interspersed with runs of fast (10-15 Hz) activity. The intracellular aspects of this complex pattern in different types of neocortical neurons are described in the following paper. Complete decortication abolished the seizure, leaving intact thalamic spindles. Injections of bicuculline in the cortex of athalamic cats resulted in similar components as those occurring with an intact thalamus. 3) Injection of bicuculline in the thalamus decreased the frequency of barbiturate spindles and increased the synchrony of spike bursts fired by thalamocortical and thalamic reticular cells but did not induce seizures. Decortication did not modify the effects of bicuculline injection in the thalamus. Our results indicate that the minimal substrate that is necessary for the production of seizures consisting of SW/PSW complexes and runs of fast activity is the neocortex.

Animals↗

PubMed1997

Block of transmitter release by botulinum C1 action on syntaxin at the squid giant synapse.

Electrophysiological, morphological, and biochemical approaches were combined to study the effect of the presynaptic injection of the light chain of botulinum toxin C1 into the squid giant synapse. Presynaptic injection was accompanied by synaptic block that occurred progressively as the toxin filled the presynaptic terminal. Neither the presynaptic action potential nor the Ca2+ currents in the presynaptic terminal were affected by the toxin. Biochemical analysis of syntaxin moiety in squid indicates that the light chain of botulinum toxin C1 lyses syntaxin in vitro, suggesting that this was the mechanism responsible for synaptic block. Ultrastructure of the injected synapses demonstrates an enormous increase in the number of presynaptic vesicles, suggesting that the release rather than the docking of vesicles is affected by biochemical lysing of the syntaxin molecule.

Animals↗

PubMed1997

Spatiotemporal patterns of spindle oscillations in cortex and thalamus.

Spindle oscillations (7-14 Hz) appear in the thalamus and cortex during early stages of sleep. They are generated by the combination of intrinsic properties and connectivity patterns of thalamic neurons and distributed to cortical territories by thalamocortical axons. The corticothalamic feedback is a major factor in producing coherent spatiotemporal maps of spindle oscillations in widespread thalamic territories. Here we have investigated the spatiotemporal patterns of spontaneously occurring and evoked spindles by means of multisite field potential and unit recordings in intact cortex and decorticated animals. We show that (1) spontaneous spindle oscillations are synchronized over large cortical areas during natural sleep and barbiturate anesthesia; (2) under barbiturate anesthesia, the cortical coherence is not disrupted by transection of intracortical synaptic linkages; (3) in intact cortex animals, spontaneously occurring barbiturate spindle sequences occur nearly simultaneously over widespread thalamic territories; (4) in the absence of cortex, the spontaneous spindle oscillations throughout the thalamus are less organized, but the local coherence (within 2-4 mm) is still maintained; and (5) spindling propagation is observed in intact cortex animals only when elicited by low intensity cortical stimulation, applied shortly before the initiation of a spontaneous spindle sequence; propagation velocities are between 1 and 3 mm/sec, measured in the anteroposterior axis of the thalamus; increasing the intensity of cortical stimulation triggers spindle oscillations, which start simultaneously in all leads. We propose that, in vivo, the coherence of spontaneous spindle oscillations in corticothalamic networks is attributable to the combined action of continuous background corticothalamic input initiating spindle sequences in several thalamic sites at the same time and divergent corticothalamic and intrathalamic connectivity.

Anesthesia↗

PubMed1997

State-dependent fluctuations of low-frequency rhythms in corticothalamic networks.

We have studied the variations in the degree of correlated firing within the low-frequency sleep rhythms (< 15 Hz) between cortical, thalamic reticular and thalamocortical neurons during changes in the amplitude and frequency of brain electrical activity in anaesthetized cats. Extracellular discharges of neuronal groups of two to five physiologically identified cortical and thalamic units were recorded simultaneously with independent microelectrodes. The firing patterns and the temporal correlation between spike-trains were evaluated by auto- and crosscorrelograms. Although the animals were under deep anaesthesia, additional doses of the same or different anaesthetics were able to alter the electroencephalographic pattern, inducing waves with higher amplitude. Similar transitions occurred spontaneously. We found that the presence of rhythmic behaviour in cells of corticothalamic networks, as well as their degree of correlated firing, was extremely sensitive to even slight alterations in the state of the electroencephalogram. Cells belonging to the same functional system, but located distantly, became highly synchronized upon the increased amplitude of brain waves. Thus, an electroencephalogram characterized by slow waves corresponds to a state of rhythmic and correlated firing among cortical and thalamic neurons. The highly coherent activity during sleep patterns transcends the borders which limit the functioning during the waking brain.

Anesthetics, General↗

PubMed1997

Synchronization of low-frequency rhythms in corticothalamic networks.

We have investigated the degree of synchronization between cortical, thalamic reticular and thalamocortical neurons of cats during low-frequency (< 15 Hz) sleep-like oscillations, as they appear under anaesthesia. We have also studied the effects exerted by cortical stimulation on the synchronization among thalamic units. Parallel experiments [Steriade et al. (1996) J. Neurosci. 16, 392-417] in this laboratory have demonstrated the similarity between the slow oscillation (< 1 Hz) under ketamine-xylazine anaesthesia and that occurring during the natural state of resting sleep. Spontaneous activity was recorded simultaneously, with independent microelectrodes, from groups of two to five physiologically identified neurons. The rhythmicity of spontaneous activity and the temporal relations between cellular discharges were statistically evaluated by auto- and crosscorrelation techniques. We have found no topography in the distribution of synchronization between thalamic reticular and thalamocortical cells. Only the slow, cortical-generated oscillation (< 1 Hz) displayed a stable frequency and correlation among groups of cortical and thalamic cells. The other two sleep oscillations (thalamic-generated spindles at 7-14 Hz and clock-like delta at 1-4 Hz) fluctuated in frequency and the degree of correlation between neurons varied. Cortical volleys entrained and synchronized thalamic cells, and triggered synchronized spindling in the thalamus. These results extend for large populations of cortical and thalamic neurons the phase relations found in intracellular recordings.

Animals↗