Biocompatibility of new nanostructural materials based on active silicate systems and hydroxyapatite: in vitro and in vivo study.

AIM: To evaluate in vitro cytotoxicity and in vivo inflammatory response to new nanostructural materials based on active calcium silicate systems (CS) and hydroxyapatite (HA-CS). METHODOLOGY: Cytotoxicity of eluates of new nanostructural noncommercial materials CS and HA-CS, and MTA (White MTA, Angelus(®) Soluções Odontológicas, Londrina, Brazil) as a control, were tested using the MTT assay on MRC-5 cells. Eluates of set materials were tested in 100% and 50% concentrations, 24 h, 7 days and 21 days post-elution. The pH values were determined for undiluted eluates of set materials. Polyethylene tubes containing the test materials (CS, HA-CS, MTA) were implanted in subcutaneous tissue of Wistar rats. Histopathological examinations were conducted at 7, 15, 30 and 60 days after the implantation. Data were statistically analyzed using three-way and one-way anova Tukey's post hoc test as well as Kruskall-Wallis test with Dunn's post hoc test at α = 0.05. RESULTS: All materials significantly reduced cell viability; especially when undiluted eluates were used (P < 0.001). After 24 h elution, cell viability was 10 ± 1.8%, 49.5 ± 4.2% and 61 ± 7.4%, for MTA, and HA-CS, respectively. However, CS and HA-CS were significantly less toxic than the control material MTA (P < 0.05). Cytotoxicity could be at least partially attributed to pH kinetics over time. Dilution of eluates of all tested materials resulted in better cell survival. Histopathological examination indicated similar inflammatory reaction, vascular congestion and connective tissue integrity associated with CS, HA-CS and MTA at each observation period (P > 0.05). The only significant difference was found for capsule thickness, that is thicker capsule was associated with HA-CS compared to MTA at 60 days (P = 0.0039). HA-CS induced moderately thick capsules (median score 3, score range 2-3), whereas MTA resulted in thin capsule formation (median score 2, score range 1-3). CONCLUSIONS: Evaluation of cytotoxicity and inflammatory response indicated better biocompatibility of CS and HA-CS, in comparison with MTA (White MTA, Angelus(®) Soluções Odontológicas, Londrina, Brazil).

New type of single chain magnet: pseudo-one-dimensional chain of high-spin Co(II) exhibiting ferromagnetic intrachain interactions.

Two new six-coordinated high-spin Co(II) complexes have been synthesized through the reactions of Co(II) salts with dipyridylamine (dpamH) and 5-nitro-salicylaldehyde (5-NO2-saloH) or 3-methoxy-salicylaldehyde (3-OCH3-saloH) under argon atmosphere: [Co(dpamH)2(5-NO2-salo)]NO3 (1) and [Co(dpamH)2(3-OCH3-salo)]NO3·1.3 EtOH·0.4H2O (2). According to the crystal packing of compound 1, two coordination cations are linked with two nitrate anions into a cyclic dimeric arrangement via N-H···O and C-H···O hydrogen bonds. In turn, these dimers are assembled into (100) layers through π-π stacking interactions between inversion-center related pyridine rings of the dpamH ligands. The crystal packing of compound 2 reveals a 1D assembly consisting solely from the coordination cations, which is formed by π-π stacking interactions between pyridine rings of one of the dpamH along the [010] and another 1D assembly of the coordination cations and nitrate anions through the N-H···O hydrogen-bonding interactions along the [001] direction. All complexes were magnetically characterized, and a new approximation method was used to fit the magnetic susceptibility data in the whole temperature range 2-300 K on the basis of an empirical expression which allows the treatment of each cobalt(II) ion in axial symmetry as an effective spin S(eff) = 1/2. In zero-field, dynamic magnetic susceptibility measurements show slow magnetic relaxation below 5.5 K for compound 2. The slow dynamics may originate from the motion of broad domain walls and is characterized by an Arrhenius law with a single energy barrier Δr/k(B) = 55(1) K for the [10-1488 Hz] frequency range. In order to reveal the importance of the crystal packing in the SCM behavior, a gentle heating process to 180 °C was carried out to remove the solvent molecules. The system, after heating, undergoes a major but not complete collapse of the network retaining to a small percentage its SCM character.

Clinical reproducibility of three electronic apex locators.

AIM: To compare the reproducibility of three electronic apex locators (EALs), Dentaport ZX, RomiApex A-15 and Raypex 5, under clinical conditions. METHODOLOGY: Forty-eight root canals of incisors, canines and premolars with or without radiographically confirmed periapical lesions required root canal treatment in 42 patients. In each root canal, all three EALs were used to determine the working length (WL) that was defined as the zero reading and indicated by 'Apex', '0.0' or 'red square' markings on the EAL display. A new K-file of the same size was used for each measurement. The file length was fixed with a rubber stop and measured to an accuracy of 0.01 mm. Measurements were undertaken by two calibrated operators. Differences in zero readings between the three EALs in the same root canal were statistically analysed using paired t-tests with the Bonferroni correction, Bland-Altman plot and Linn's concordance correlation coefficients at α = 0.05. RESULTS: Mean and standard deviation values measured by the three EALs showed no statistically significant differences. Identical readings by all three EALs were found in 10.4% of root canals. Forty-three per cent of readings differed by less than ± 0.5 mm and 31.3% exceeded a difference of ± 1 mm. CONCLUSIONS: The clinical reproducibility of Dentaport ZX, RomiApex A-15 and Raypex 5 was confirmed with the majority of readings within the ± 1.0 mm range. However, the small number of identical zero readings suggests that EALs are not reliable as the sole means of WL determination under clinical conditions.

Identification of distinct C3b and C4b recognition sites in the human C3b/C4b receptor (CR1, CD35) by deletion mutagenesis.

Complementary DNA clones encoding the NH2-terminal region of human CR1 have been isolated and sequenced. The deduced complete amino acid sequence of the F allotype of human CR1 contains 2,039 residues, including a 41-residue signal peptide, an extracellular domain of 1,930 residues, a 25-amino acid transmembrane domain, and a 43-amino acid cytoplasmic region. The extracellular domain is composed exclusively of 30 short consensus repeats (SCRs), characteristic of the family of C3/C4-binding proteins. The 28 NH2-terminal SCRs are organized as four long homologous repeats (LHRs) of seven SCRs each. The newly sequenced LHR, LHR-A, is 61% identical to LHR-B in the NH2-terminal two SCRs and greater than 99% identical in the COOH-terminal five SCRs. Eight cDNA clones were spliced to form a single construct, piABCD, that contained the entire CR1 coding sequence downstream of a cytomegalovirus promoter. COS cells transfected with piABCD transiently expressed recombinant CR1 that comigrated with the F allotype of erythrocyte CR1 on SDS-PAGE and that mediated rosette formation with sheep erythrocytes bearing C4b and C3b. Recombinant CR1 also had factor I-cofactor activity for cleavage of C3(ma). Analyses of six deletion mutants expressed in COS cells indicated that the NH2-terminal two SCRs of LHR-A contained a site determining C4 specificity and the NH2-terminal two SCRs of LHR-B and -C each had a site determining C3 specificity. The presence of these three distinct sites in CR1 may enable the receptor to interact multivalently with C4b/C3b and C3b/C3b complexes generated during activation of the classical and alternative pathways.

Complement-immunoglobulin interactions.

Normal circulating immunoglobulin may control complement binding to targets and thereby the manifestations of autoimmune disease. Molecular analysis of IgG and IgM mutants suggests that C1q binding by IgG utilizes a core Glu-X-Lys-X-Lys motif (where X is any amino acid). Additional amino acids, particularly homologous proline residues at position 331 in IgG and 436 in IgM, appear critical for classical pathway initiation. Glycosylation of IgG heavy chain is important in C1q binding, as well as glycosylation of IgA heavy chain for alternative pathway initiation. Additional recent evidence suggests an important role for C3 in antigen presentation. The data also raise the possibility that C3 plays a significant role in the intracellular antigen processing pathway.

Evaluation of Staining-Dependent Colour Changes in Resin Composites Using Principal Component Analysis.

Colour changes in Gradia Direct™ composite after immersion in tea, coffee, red wine, Coca-Cola, Colgate mouthwash, and distilled water were evaluated using principal component analysis (PCA) and the CIELAB colour coordinates. The reflection spectra of the composites were used as input data for the PCA. The output data (scores and loadings) provided information about the magnitude and origin of the surface reflection changes after exposure to the staining solutions. The reflection spectra of the stained samples generally exhibited lower reflection in the blue spectral range, which was manifested in the lower content of the blue shade for the samples. Both analyses demonstrated the high staining abilities of tea, coffee, and red wine, which produced total colour changes of 4.31, 6.61, and 6.22, respectively, according to the CIELAB analysis. PCA revealed subtle changes in the reflection spectra of composites immersed in Coca-Cola, demonstrating Coca-Cola's ability to stain the composite to a small degree.

Bulbospinal serotoninergic pathways in the frog Rana pipiens.

We combined retrograde fluorescent tracing with rhodamine immunofluorescence to identify the origin of serotoninergic neurons with descending projections to the spinal cord of frogs. After injections of Fluoro-gold into the spinal cord, retrogradely labeled immunoreactive serotoninergic neurons were detected in the caudal part of the brainstem from the level of the obex through the level of the VIII nerve. These doubly labeled cells were distributed along the midline throughout the rostrocaudal extent of the dorsal portion of the raphe nuclear region. Doubly labeled neurons were more numerous in the rostral than in the caudal part of the raphe area. The fluorescent tracer 1,1'-dioctadecyl-3,3,3'3'-tetramethylindocarbocyanine perchlorate (DiI) was then placed in and around the middle and rostral raphe nuclear region. Anterogradely labeled fibers could be traced bilaterally in the lateral portion of the dorsal funiculus and the lateral and ventral funiculi. These fibers were seen terminating in the dorsal and ventral horns, as well as in the intermediate grey matter. After placement of DiI in the caudal raphe area, labeled fibers were found only in the intermediate grey and ventral horn. These findings suggest that the organization of bulbospinal serotoninergic pathways in the frog is similar to that of mammals, and that an isolated amphibian spinal cord preparation could be a useful model for pharmacological and physiological studies of the action of serotonin (5HT) in the spinal cord.

Retrograde tracing of projections between the nucleus submedius, the ventrolateral orbital cortex, and the midbrain in the rat.

The fluorescent tracers fluoro-gold and 1,1'-dioctadecyl-3,3,3,3-tetramethyl indocarbocyanine perchlorate were used as retrograde markers to examine reciprocal connections between the rat nucleus submedius and the ventrolateral orbital cortex. In addition, midbrain projections to each of these regions were examined. In the prefrontal cortex, we found that input from the nucleus submedius terminates rostrally within the lateral and ventral areas of the ventrolateral orbital cortex. Conversely, the cortical input to the nucleus submedius originates from the medial and dorsal parts of the ventrolateral orbital cortex. Our data also demonstrated that neurons from the ventrolateral periaqueductal gray and the raphe nuclei project to the midline nuclei of the thalamus, including a small projection to the nucleus submedius. We further determined that regions within the ventrolateral periaqueductal gray and raphe nuclei project to the ventrolateral orbital cortex, and that these regions overlap with those that project to the nucleus submedius. These findings suggest that the nucleus submedius might be part of a neural circuit involved in the activation of endogenous analgesia.

A morphological study of cat dorsal spinocerebellar tract neurons after intracellular injection of horseradish peroxidase.

This work represents an attempt to elucidate structural features of electrophysiologically characterized, individual cat dorsal spinocerebellar tract (DSCT) neurons by using intracellular application of horseradish peroxidase (HRP). Intracellular recordings and HRP injections were made in DSCT neurons of the Clarke's column in cat lumbar (L3) spinal cord. The units were identified by antidromic invasion following electrical stimulation of the ipsilateral dorsolateral funiculus at C1. In addition, sensory inputs to the DSCT neurons were determined by natural (adequate) stimuli applied to the hind limb with intact innervation. The morphological analysis is based on data obtained from 19 well-stained electrophysiologically identified neurons located in Clarke's column. Thirteen of these units received excitatory sensory inputs from muscle receptors, two were activated by cutaneous afferents only, and four had a convergent (muscle + cutaneous) input. The DSCT--muscle cells were equivalent to the large Clarke cells (class C of Leowy, '70). Their dendrites were oriented primarily in the rostro--caudal direction (up to 2500 micron) and appeared generally smooth except for some branchlets. In four of these cells, the axon was traced into the lateral funiculus. In light microscopic analysis there was no evidence that axon collaterals arose from these axons during the initial trajectory through the spinal grey matter. The four DSCT--convergent neurons were similar in shape to the DSCT--muscle units although they appeared to have somewhat smaller cell bodies. Of the two DSCT--cutaneous neurons one was found to be of the B type, with the dendritic tree having fewer branches and oriented mainly in the medio--lateral direction. The other cell, however, turned out to be similar in appearance to the C type Clarke neurons.

Serotoninergic axonal contacts on identified cat spinal dorsal horn neurons and their correlation with nucleus raphe magnus stimulation.

This study examined the distribution of serotoninergic (5-HT) immunoreactive axonal contacts on spinal laminae I and II neurons by combining the intracellular horseradish peroxidase (HRP) method with immunocytochemistry. In addition, the 5-HT distribution was correlated with effects produced by electrical stimulation within the nucleus raphe magnus (NRM). Responses of lamina I neurons and lamina II stalked cells to noxious stimulation were markedly suppressed during NRM stimulation. In contrast, responses of nociceptive lamina IIa islet or non-nociceptive lamina IIb islet cells remained unchanged during nucleus raphe magnus stimulation. These inhibitory influences were positively correlated with the distribution of 5-HT immunoreactive contacts on these neurons. Nociceptive lamina I neurons and lamina II stalked cells received a significantly greater number of contacts (average of 74 and 63, respectively) than either nociceptive lamina IIa islet or non-nociceptive lamina IIb islet cells (average of 25 and eight contacts, respectively). Irrespective of cell type, most 5-HT contacts occurred on dendritic shafts rather than spines. These data reveal a differential distribution of 5-HT contacts on neurons in spinal laminae I and II, and indicate that at least a portion of the NRM modulation of dorsal horn neuronal activity is serotoninergic and concentrated on the dendritic shafts of nociceptive lamina I neurons and lamina II stalked cells.

Immunoglobulin in the control of complement action.

Complement is a critical element of innate immunity, protecting individuals from a wide variety of microbial infections. This group of proteins is responsible for many features of inflammation and tissue damage. Because of its ability to mediate autoimmune tissue damage and to destroy host tissues, it is under tight regulation with many circulating and cell-membrane-bound complement regulatory proteins. The function of much of the circulating immunoglobulin has never been defined. We have advanced the hypothesis that one function of circulating immunoglobulin is to down-regulate complement attack on host tissues in the presence of anti-self antibody. The data to support this hypothesis are reviewed. The data are consistent with the suggestion that one mechanism of action of intravenous immunoglobulin, used to treat patients with a variety of autoimmune diseases, is prevention of complement-mediated attack on host tissues.

Long-term changes in sciatic-evoked A-fiber dorsal horn field potentials accompany loose ligation of the sciatic nerve in rats.

The goal of the present study was to examine whether loose ligation of the sciatic nerve was associated with long-term changes in neuronal excitability in the spinal dorsal horn in urethane-anesthetized rats. The sciatic nerve was stimulated with 0. 1 ms long pulses at 1 stimulus/5 min, and the evoked dorsal horn field potentials remained stable in the absence of tetanic stimulation. In one set of control and ligated animals, high-frequency tetanic stimulation was applied to the nerve at 50 Hz (one 400 ms train of twenty 0.1 ms pulses), and the field potentials were recorded again (1 stimulus/5 min) for up to 4 h post-tetanus. In control animals, this protocol produced significant increases in field potential amplitudes at 15, 30 and 60 min post-tetanus. Interestingly, after this time the evoked field potentials began to decrease, and attained less than 50% of their pre-tetanic values at 240 min post-tetanus. In contrast, in ligated rats the pattern of post-tetanic potentiation was significantly different as the increases in amplitude persisted, and at 240 min post-tetanus the field potentials were almost twice their baseline values. In another set of control and ligated animals, low-frequency tetanic stimulation was applied at 5 Hz (one 400 ms train of two 0.1 ms pulses). Again a differential pattern of post-tetanic responses between control and ligated rats was seen. In control animals, a significant decrease in amplitude was evident within 30 min, and the depression became progressively more pronounced as the field potentials attained about a quarter of their baseline values at 180 min, and remained at these low levels at 240 min post-tetanus. On the other hand, in ligated animals, the depression was not significant, and at 240 min post-tetanus the field potentials were still at about 80% of their baseline values. These data demonstrate that long-term changes in spinal dorsal horn neuronal excitability accompany sciatic ligation to perhaps contribute to the development of neuropathic pain. These changes may result from a lessening of normally strong inhibitory processes in the spinal dorsal horn to generate conditions which favor post-tetanic potentiation over depression of dorsal horn neuronal responses.

Contribution of GABA-A receptors to metaplasticity in the spinal dorsal horn.

Metaplasticity is a higher-order form of synaptic plasticity that is induced by synaptic or cellular activity, which by itself may not produce changes in synaptic strength, but which modifies subsequent changes in synaptic efficacy. In this description of metaplasticity in the spinal dorsal horn, we report that a 50 Hz high-frequency tetanus, previously shown to elicit a potentiation of sciatic-evoked A-fiber spinal dorsal horn potentials, caused a depression when coupled with a more rapid rate of repetitive stimulation. This depression appeared to be dependent upon GABA(A) receptor activation because the 50 Hz tetanus elicited a persistent potentiation when the GABA(A) antagonist bicuculline was injected at 1 mg/kg (but not at 0.5 mg/kg) prior to tetanic stimulation. These data suggest the presence of strong inhibitory inputs in the spinal dorsal horn that are activated by an increased rate of primary afferent firing. The activation of these inputs may be necessary to prevent prolonged bursts of afferent activity from modifying synaptic strength because the latter may contribute to the development of persistent pain following peripheral nerve injury.

A new qualitative method for detecting IgD in unconcentrated cerebrospinal fluid.

We report a method which is capable of demonstrating the isoelectric focusing (IEF) pattern of immunoglobulin D in unconcentrated cerebrospinal fluid (CSF) samples containing as little as 0.1-0.5 ng of total IgD. The method used was an immuno-sandwich technique, with alkaline phosphatase enzyme amplification. Oligoclonal and polyclonal IgD patterns were seen in CSF samples. No cross-reactivity with other immunoglobulins (IgG, IgA and IgM) was detected.

Diverse actions of 5-hydroxytryptamine on frog spinal dorsal horn neurons in vitro.

The effects of 5-hydroxytryptamine on the membrane potential and input resistance of 86 dorsal horn neurons were studied using intracellular recordings in isolated, hemisected spinal cords of adult frogs (Rana pipiens). Bath application of serotonin (5-100 microM) caused membrane depolarizations in 58 (67%) neurons, hyperpolarizations in 12 (14%) cells, biphasic responses in nine (11%) neurons, and no detectable change in seven (8%) cells. In some neurons depolarized by serotonin, the amine's responses could be mimicked by the selective 5-HT2 agonist (+/-)-1(2,5-dimethoxy-4-iodophenyl)-2-aminopropane hydrochloride and the 5-HT1C/2 agonist alpha-methyl-5-hydroxytryptamine, and blocked by the 5-HT1C/2 antagonists ketanserin and mianserin. In other neurons depolarized by serotonin, the 5-HT3 agonist 2-methyl-5-hydroxytryptamine mimicked, and the 5-HT3 antagonist, 3-tropanyl-3,5-dichlorobenzoate, blocked the serotonin-induced responses. Depolarizing responses due to activation of 5-HT1C/2 receptors were generally accompanied by increases in the membrane input resistance, whereas depolarizations mediated by 5-HT3 receptors were associated with a decreased membrane input resistance. Superfusion with tetrodotoxin or low-Ca2+/high-Mg(2+)-containing media abolished about half of the depolarizing responses. Hyperpolarizations caused by serotonin were associated with a decrease in membrane input resistance, and might have been due to activation of a potassium conductance. These responses persisted in bathing solutions containing tetrodotoxin or low-Ca2+/high-Mg2+. The 5-HT1A agonist 8-hydroxy-2-(di-N-propylamine)tetralin hydrobromide mimicked, whereas the 5-HT1A antagonist spiroxatrine blocked, these hyperpolarizing responses. Other antagonists selective for 5-HT1C/2 or 5-HT3 receptors were without effect. Serotonin-produced biphasic responses consisted of either an initial depolarization followed by a hyperpolarization or the reverse. The selective 5-HT2 agonist (+/-)-1(2,5-dimethyoxy-4-iodophenyl)-2-aminopropane hydrochloride could only mimic the depolarizations, whereas the 5-HT1A agonist 8-hydroxy-2-(di-N-propylamine)tetralin hydrobromide produced only the hyperpolarizations. Spiroxatrine, a 5-HT1A antagonist, blocked only the hyperpolarizations without affecting the depolarizations, and methysergide, a non-specific 5-HT receptor antagonist, depressed both the depolarizations and hyperpolarizations. Serotonin also appeared to affect spinal dorsal horn neurons indirectly because it produced excitatory postsynaptic potentials, inhibitory postsynaptic potentials, and a mixture of both.(ABSTRACT TRUNCATED AT 400 WORDS)

Responses of type A cat spinal ganglion neurons to repetitive stimulation of their central and peripheral processes.

Intracellular recordings were made from L7-S1 type A spinal ganglion neurons of anesthetized cats while electrical stimulation was delivered repetitively to their associated dorsal root and the sciatic nerve. The general response pattern of these neurons changed during stimulation at progressively higher rates. The changes were observable as jitter in onset latency of the evoked spikes, inability of evoked responses to follow electrical stimuli in a 1:1 manner (spike failure), reduction in action potential amplitude, and decomposition of the full spike into its non-myelinated and myelinated components. The frequency following ability of these spike components was in the order of full spike less than non-myelinated less than myelinated. In jitter in onset latency and inability to follow high frequency stimulation was determined only for the full spike, as is typical for antidromicity criteria, a wide frequency following spectrum was obtained for our sample of spinal ganglion neurons. Less than a third of the cells were able to follow stimulation rates in excess of 200 Hz, and about a fifth of the neurons failed to follow any rates greater than 20 Hz. Most of the neurons activated from both the dorsal root and sciatic nerve responded with the same pattern of stimulus-evoked responses. However, some of these cells exhibited strikingly different patterns to dorsal root and sciatic stimulation, including the presence of prepotentials following stimulation of one, but not the other, process. These prepotentials occurred in the depolarizing direction, at threshold stimulation were all-or-none in nature, generated spikes that varied in onset latency, and failed to occur at even low-to-moderate rates of stimulation. The results indicate that the frequency following spectrum of cat type A ganglion neurons is wide, and that it is their somata that are most vulnerable to high frequency stimulation. It is possible that some of the observed prepotentials are functional manifestations of synaptic contacts in spinal ganglia.

A laser nephelometric method for measuring alternative pathway complement activity of human and murine serum.

A kinetic method for the evaluation of alternative pathway complement activity in man and mice is presented. A laser nephelometer was employed for detection of non-sensitized rabbit erythrocyte lysis based on the observation that the intensity of laser scatter (LS) is proportional to the number of erythrocytes in suspension. During erythrocyte lysis a continuous decline in LS is observed since lytic products do not evoke LS. Utilizing the indirect Coombs test and cross-electrophoresis it was shown that rabbit erythrocyte lysis causes activation of the alternative complement pathway. This method is modified for room temperature, is independent of sample hemoglobin content and, in its micro version, it can be done with 10 microliter of human serum, i.e. 50 microliters of murine serum.

Antibodies against rabbit red blood cells in murine serum and their effect on the activity of complement alternative pathway.

Antibody content against rabbit red blood cells (anti-RaRBC) in murine sera of different strains (Swiss, CBA, C57BL/6, AKR, BALB/c) and activity of complement alternative pathway (AP) were investigated. In contrast to the CBA and C57BL/6, random-bred Swiss strain and inbred BALB/c and AKR strains are good producers of these natural antibodies. There is no correlation between AP activity and anti-RaRBC content. Isolated human anti-RaRBC antibodies, IgM and IgG classes, lead to the enhancement of APhu and APmo activity, contrary to the murine anti-RaRBC which belong solely to IgM class, and do not express this capability.

Increased spinal monoamine concentrations after chronic thoracic dorsal rhizotomy in goats.

In goats, bilateral thoracic dorsal rhizotomy (TDR) causes severe ventilatory failure during exercise, followed by progressive functional recovery. We investigated spinal neurochemical changes associated with TDR and/or functional recovery by measuring spinal concentrations of the monoamines serotonin (5-HT), norepinephrine, and dopamine via HPLC. Changes in 5-HT and calcitonin gene-related peptide were visualized with immunohistochemistry. Goat spinal cords were compared 4-15 mo after TDR from T(2) to T(12) (n = 7) with sham-operated (n = 4) or unoperated controls (n = 4). TDR increased the concentration of cervical 5-HT (C(5)-C(6); 122% change), caudal thoracic norepinephrine (T(7)-T(11); 53% change), and rostral thoracic dopamine (T(3)-T(6); 234% change). TDR increased 5-HT-immunoreactive terminal density (dorsal and ventral horns) and nearly eliminated calcitonin gene-related peptide immunoreactivity in the superficial laminae of the dorsal horn in rostral thoracic segments; both effects became less pronounced in caudal thoracic segments. Thus TDR elevates monoamine concentrations in discrete spinal regions, including possible compensatory changes in descending serotonergic inputs to spinal segments not directly affected by TDR (i.e., cervical) but associated with functionally related motor nuclei (i.e., phrenic nucleus).

Symposium overview: chemical modulation of neuroreceptors and channels via intracellular components.

Whereas the roles of G proteins and protein kinases in various neuroreceptors and ion channels have been studied extensively, their roles in the actions of drugs and toxicants on these receptors and channels remain to be elucidated. Almost all drugs and toxicants exert multiple actions on multiple target sites, and there is no reason to assume that a chemical modulates a receptor/channel via a single mechanism. In fact, experimental evidence is slowly but steadily being accumulated to indicate that certain drugs and toxicants modulate neuroreceptor/channel functions through interactions with intracellular components such as G proteins and protein kinases. Multiple actions of a toxicant on various receptors/channels may be explained on the basis of its interaction with the G protein/kinase system that is a common denominator of the target sites. This is a virgin field that promises a quantum leap in the coming years. Each presentation and discussion will focus on expected future developments and potential significance in the field of neurotoxicology.