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Biomedical subjects

R J Miller

Publications and source records attributed to R J Miller.

At least 163 records · Page 9Linked to original sources
PubMed1993

Calcium buffering properties of calbindin D28k and parvalbumin in rat sensory neurones.

1. We have examined the ability of the Ca(2+)-binding proteins (CABP) calbindin D28k and paravalbumin to modulate increases in the intracellular free Ca2+ concentration ([Ca2+]i), produced by brief depolarizations, in rat dorsal root ganglion (DRG) neurones. 2. In order to obtain good voltage control, we replated DRG neurones prior to performing these experiments. Immunocytochemical staining of these cells revealed that approximately 10% stained for CABPs. 3. Using fluorescently labelled parvalbumin, we demonstrated that in the whole-cell voltage clamp mode the protein freely entered the cell soma with a mean half-life t0.5 of 6 min 22 s +/- 54 s. 4. Analysis of the effects of calbindin D28k (370 microM) and parvalbumin (1 mM) on Ca2+ currents in the whole-cell voltage clamp mode, revealed that neither protein changed the rate of inactivation of the Ca2+ current or its rate of run-down. 5. Introducing either calbindin D28k (370 microM) or parvalbumin (1 mM) into the cell soma did not significantly alter the basal [Ca2+]i when compared to control cells. 6. Compared to control cells, both CABPs significantly reduced the peak [Ca2+]i obtained for a Ca2+ influx of an equivalent charge density, whereas lysozyme (1 mM), a protein with low affinity for Ca2+, failed to do so. 7. Calbindin D28k caused an 8-fold decrease in the rate of rise in [Ca2+]i and altered the kinetics of decay of [Ca2+]i to a single slow component. Parvalbumin also slowed the rate of rise in [Ca2+]i. Parvalbumin selectively increased a fast component in the decay of the Ca2+ signal. 8. These data demonstrate that both calbindin D28k and paravalbumin effectively buffer Ca2+ in a cellular environment and may therefore regulate Ca(2+)-dependent aspects of neuronal function.

Animals↗
PubMed1993

Metabotropic glutamate receptors depress afferent excitatory transmission in the rat nucleus tractus solitarii.

1. We have previously demonstrated that the metabotropic glutamate receptor (mGluR) agonist (1S,3R)-1-aminocyclopentane-1,3-dicarboxylate [(1S;3R)-ACPD] presynaptically inhibits evoked glutamatergic excitatory postsynaptic currents (EPSCs) in patch-clamped rat nucleus tractus solitarius (NTS) neurons recorded in thin slices. The present study investigated the ability of endogenously released glutamate to modulate EPSCs in the NTS. 2. A low-frequency tetanus of the tractus solitarius (TS) resulted in either posttetanic potentiation (PTP) (8 of 21 cells) or depression (13 of 21 cells) of monosynaptic EPSCs recorded in the presence of D(-)2-amino-5-phosphonopentanoic acid (AP5) and bicuculline. 3. The amplitude of the EPSC was not significantly affected by the bath application of the mGluR antagonist (+) alpha-methyl-4-carboxyphenylglycine (MCPG). 4. In the presence of MCPG, a low-frequency tetanus resulted in PTP of the EPSC in all neurons. PTP was significantly enhanced in those cells previously exhibiting PTP. 5. The results suggest that presynaptic mGluRs on TS projections to the NTS may be activated by endogenously released glutamate at physiologically relevant stimulus frequencies and therefore play a role in the modulation of autonomic afferent transmission.

Afferent Pathways↗
PubMed1993

Profiles and predictors of assaultiveness for different psychiatric ward populations.

OBJECTIVE: The authors examined associations between patient-related characteristics and assaultiveness on six different psychiatric wards to determine 1) the relative contributions of demographic, disorder-related, and diagnostic variables to prediction of assaultiveness and 2) how ward composition and type of victim affect prediction of assaultiveness. METHOD: Hospital records of 1,025 inpatients residing in psychiatric wards within a 6-month time frame were reviewed for evidence of assaultiveness. Data on all 260 assaultive patients and a sample of 136 of the nonassaultive patients were analyzed with multiple regression to predict assaultiveness scores for each ward and each victim category. RESULTS: The findings indicated high rates of assaultive patients and assaults on fellow patients. Multiple regression results produced different predictors of assaultiveness for different wards but not for different categories of assault victim within each ward. Overall, age and sex consistently failed to predict assaultiveness, whereas greater assaultiveness was significantly associated with a greater proportion of time hospitalized since first admission. The most powerful unique predictors of assaultiveness scores were diagnostic distinctions derived from data on coexistent diagnoses. The most assaults were by acute patients whose diagnoses excluded organic mental disorder but included either bipolar disorder or personality disorder and longer-stay patients whose exclusive diagnosis was organic mental disorder. CONCLUSIONS: This identification, albeit modest, of risk factors for assaultiveness on different wards nevertheless provides information fundamental to the management policies of psychiatric institutions. The findings caution against aggregating different ward populations for research on assaultiveness and endorse the usefulness of coexistent diagnoses for predicting assaultiveness.

Bipolar Disorder↗
PubMed1993

Activation of metabotropic glutamate receptors produces reciprocal regulation of ionotropic glutamate and GABA responses in the nucleus of the tractus solitarius of the rat.

Whole-cell voltage-clamp recordings were made in thin transverse slices from neurons of the dorsomedial subdivision of the nucleus of the tractus solitarius (NTS) of the rat. Cells were exposed to either the ionotropic glutamate receptor agonist (R, S)-alpha-amino-3-hydroxy-5-methylisoxazole-4-propionate (AMPA) or the GABAA receptor agonist muscimol via pressure ejection directed at the cell soma. The metabotropic glutamate receptor agonist 1S,3R-1-aminocyclopentane-1,3-dicarboxylate (1S,3R-ACPD; 2-100 microM) reversibly depressed muscimol-evoked currents. Conversely, 1S,3R-ACPD reversibly potentiated AMPA-evoked currents. High-frequency stimulation of the tractus solitarius in the presence of 6,7-dinitroquinoxaline-2,3-dione and D-2-amino-5-phosphonopentanoic acid also produced a reversible depression of muscimol-evoked currents that was occluded in the presence of 100 microM 1S,3R-ACPD. 8-Br-cGMP or brain-derived natriuretic peptide mimicked the effects of 1S,3R-ACPD on AMPA and muscimol currents. However, agents that mimicked the actions of cAMP or diacylglycerol did not. These findings indicate that metabotropic glutamate receptors may mediate multiple components of excitatory transmission in the NTS including modulation of glutamate and GABA-activated ion channels.

Animals↗
PubMed1993

Selective opioid agonists modulate afferent transmission in the rat nucleus tractus solitarius.

We examined the effects of agonists at mu, delta and kappa opioid receptors on neurons located in the nucleus tractus solitarius of the rat using whole-cell patch-clamp recordings in brainstem slices. The mu selective opioid agonist DAMGO hyperpolarized most neurons tested. This effect was associated with the activation of a K(+)-conductance. The effect of DAMGO tended to desensitize and was blocked by naloxone. Dynorphin A also produced this effect. However, the kappa-1-selective opioid agonist U-69593 and two delta-selective opioid agonists did not. DAMGO also depressed glutamate-mediated excitatory postsynaptic potentials and GABA-mediated evoked by stimulation of the tractus solitarius. Dynorphin A, U-69593 and delta-opioid agonists also reduced the excitatory postsynaptic potential, although they were less effective than DAMGO. The presynaptic inhibitory effects of DAMGO were also blocked by naloxone, but did not desensitize. These actions may help to explain the ability of opiates to modulate a variety of autonomic reflexes.

Action Potentials↗
PubMed1993

Neuropeptide Y and pancreatic polypeptide reduce calcium currents in acutely dissociated neurons from adult rat superior cervical ganglia.

We examined the effects of neuropeptide Y (NPY) and pancreatic polypeptide on calcium currents (ICa) in acutely dissociated neurons from the adult rat superior cervical ganglion. We found that NPY inhibited the ICa with an estimated IC50 value of 140 nM. This inhibitory effect appeared to be restricted to a subset of cells which were smaller in diameter than the general population. The effect of NPY on the ICa was prevented by pretreatment with pertussis toxin, suggesting the involvement of a GTP-binding protein of the Gi or Go subtype. omega-conotoxin GVIA also occluded the effects of NPY, which suggests that these were directed toward N-type Ca++ channels. The effects of NPY were mimicked by the fragment NPY (13-36) but not by peptide YY, indicating that a receptor distinct from a Y1- or a Y2-like NPY receptor was involved. Finally, we also observed that pancreatic polypeptide inhibited the ICa, suggesting that a pancreatic polypeptide receptor is also present on superior cervical ganglion neurons.

Animals↗
PubMed1993

Zinc protoporphyrin-IX blocks the effects of metabotropic glutamate receptor activation in the rat nucleus tractus solitarii.

The effects of the metabotropic glutamate receptor agonist (1S,3R)-1-aminocyclopentane-1,3-dicarboxylic acid [(1S,3R)-ACPD] on ionic current responses produced by ionotropic glutamate and gamma-aminobutyric acid (GABA)A receptor activation in the nucleus of the tractus solitarius (NTS) were examined. Recordings were made in the dorsomedial subdivision of the NTS adjacent to the area postrema in transverse brainstem slices of the rat. (1S,3R)-ACPD produced a small inward current (IACPD) associated with a decrease in conductance in approximately 50% of recordings. Monosynaptic excitatory postsynaptic currents (EPSCs) evoked by electrical stimulation in the region of the tractus solitarius in the presence of D-amino-5-phosphonopentanoic acid and bicuculline were reversibly reduced by (1S,3R)-ACPD in > 90% of cells. The inward current evoked by pressure application of alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) (IAMPA) was potentiated in the presence of (1S,3R)-ACPD, whereas the outward current evoked by the GABAA receptor agonist muscimol (IMUSC) was inhibited. We have previously demonstrated that these effects may involve the activation of soluble guanylate cyclase. The diffusible second messengers nitric oxide and carbon monoxide are known to activate soluble guanylate cyclase. The nitric oxide synthase inhibitor L-omega-nitroarginine failed to inhibit responses to (1S,3R)-ACPD. The selective heme oxygenase inhibitor Zn-protoporphyrin-IX, which would be expected to block the production of carbon monoxide, antagonized the effects of (1S,3R)-ACPD on EPSCs, IAMPA, and IMUSC. However, IACPD was not blocked. A relatively inactive metalloprotoporphyrin, Cu-protoporphyrin-IX was ineffective. A cell-permeant form of cGMP, 8-Br-cGMP inhibited EPSCs, IAMPA, and IMUSC in the presence of Zn-protoporphyrin-IX but did not induce an inward current. These results further support the hypothesis that multiple metabotropic glutamate receptors exist in the NTS, and they suggest that one of these may be coupled to the activation of a soluble guanylate cyclase via the liberation of an easily diffusible second messenger such as carbon monoxide.

Action Potentials↗
PubMed1992

Picosecond phase grating spectroscopy of hemoglobin and myoglobin: energetics and dynamics of global protein motion.

Phase grating spectroscopy has been used to follow the optically triggered tertiary structural changes of carboxymyoglobin (MbCO) and carboxyhemoglobin (HbCO). Probe wavelength and temperature dependencies have shown that the grating signal arises from nonthermal density changes induced by the protein structural changes. The material displaced through the protein structural changes leads to the excitation of coherent acoustic modes of the surrounding water. The coupling of the structural changes to the fluid hydrodynamics demonstrates that a global change in the protein structure is occurring in less than 30 ps. The global relaxation is on the same time scale as the local changes in structure in the vicinity of the heme pocket. The observed dynamics for global relaxation and correspondence between the local and global structural changes provides evidence for the involvement of collective modes in the propagation of the initial tertiary conformational changes. The energetics can also be derived from the acoustic signal. For MbCO, the photodissociation process is endothermic by 21 +/- 2 kcal/mol, which corresponds closely to the expected Fe-CO bond enthalpy. In contrast, HbCO dissipates approximately 10 kcal/mol more energy relative to myoglobin during its initial tertiary structural relaxation. The difference in energetics indicates that significantly more energy is stored in the hemoglobin structure and is believed to be related to the quaternary structure of hemoglobin not present in the monomeric form of myoglobin. These findings provide new insight into the biomechanics of conformational changes in proteins and lend support to theoretical models invoking stored strain energy as the driving force for large amplitude correlated motions.

Carboxyhemoglobin↗
PubMed1992

5-Hydroxytryptamine-3 receptors modulate synaptic activity in the rat nucleus tractus solitarius in vitro.

Whole-cell patch clamp recordings were made from neurons in the rat nucleus tractus solitarius (NTS) in transverse brainstem slices. 5-Hydroxytryptamine (5-HT, 100 microM) and the selective 5-HT3 receptor agonist 2-methyl-5-HT (2-CH3-5-HT, 100 microM) depolarized 86% of NTS neurons at resting membrane potential (Vm). This response was resistant to tetrodotoxin (TTX) and Co2+ application. In addition, 2-CH3-5-HT (500 nM-100 microM) increased the amplitude and frequency of both excitatory and inhibitory spontaneous synaptic potentials. This effect was also TTX-resistant, but was abolished by Co2+. The effects of 2-CH3-5-HT on EPSPs and IPSPs evoked by electrical stimulation of the tractus solitarius (TS) were analyzed separately in the presence of bicuculline or 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX), respectively. Concentrations of 2-CH3-5-HT between 500 nM and 1 microM decreased the amplitude of evoked EPSPs and IPSPs with similar potency. The selective 5-HT3 receptor antagonists ICS 205-930 (10 nM) and MDL 72222 (10 microM) reversibly blocked the effects of 2-CH3-5-HT at all doses examined. It is concluded that 5-HT3 receptors can mediate both pre- and postsynaptic responses in the NTS.

Animals↗
PubMed1992

Activation of endothelin receptors by sarafotoxin regulates Ca2+ homeostasis in cerebellar astrocytes.

We carried out experiments designed to investigate the effects of sarafotoxin-6B (SFTx) on [Ca2+]i in cerebellar astrocytes using the Ca2+ indicator fura-2. Both endothelin-1 and sarafotoxin-6B increased [Ca2+]i in individual cerebellar astrocytes in cell culture. The shape of the response was variable but usually consisted of an initial peak of [Ca2+]i followed by an extended plateau increase in [Ca2+]i. In Ca(2+)-free medium only the initial peak was observed. If Ca2+ was subsequently readmitted to the external medium a plateau was now formed. When external Ca2+ was removed during a plateau, [Ca2+]i rapidly declined; replacing the external Ca2+ reversed this decline. The plateau was also reversibly reduced by addition of Ni2+ (5 mM) to the external medium. Addition of 50 mM K+ produced a small increase in [Ca2+]i in most cells. This response was blocked by nimodipine. However, nimodipine only slightly blocked the plateau increase in [Ca2+]i that was formed following activation of endothelin receptors. Furthermore, perfusion of cells with 50 mM K+ during the plateau portion of a response to SFTx reduced [Ca2+]i. In some cells addition of a phorbol ester produced a sustained increase in [Ca2+]i that was blocked by nimodipine. In conclusion, activation of endothelin receptors by SFTx in cerebellar astrocytes produces both Ca2+ mobilization and Ca2+ influx. The pathway for Ca2+ influx is predominantly a non-voltage-dependent one, although some entry through a dihydropyridine-sensitive pathway also appears to occur. Furthermore, activation of protein kinase C in cerebellar astrocytes activates voltage-sensitive Ca2+ channels.

Animals↗
PubMed1992

Inhibition of tumor implantation at sites of trauma by Arg-Gly-Asp containing proteins and peptides.

We report on the inhibition of wound implantation by TA3Ha mammary carcinoma cells by Arg-Gly-Asp containing proteins and peptides using a hepatic wedge resection model. Intravenously injected TA3Ha cells rarely form tumor in the liver of syngeneic mice, but after hepatic wedge resection, 45% (107/240) of the mice develop tumors in the hepatic wound. Hepatic wound implantation is significantly (P = 0.01) inhibited by pretreating the cells with whole mouse plasma, but not with fibrinogen-depleted plasma or serum. Tumor inhibition is also achieved by pretreatment of cells with fibrinogen (P = 0.05-0.0004), fibronectin (P = 0.007) and laminin, but not by albumin. The active domain appears to be the RGDS sequence since the deca- and tetrapeptides containing RGDS inhibit wound implantation (P less than 0.05). However, the tetrapeptide Arg-Gly-Glu-Ser has no such activity. None of these agents affects ascites tumor formation by the intraperitoneally injected cells, suggesting that anchorage independent growth of cells is not affected. We propose that proteins and peptides containing RGD occupy the binding sites and prevent the cells from interacting with cell adhesion proteins in healing wounds. Proteins and/or peptides containing RGD may be useful for preventing local recurrence in postsurgical cancer patients.

Animals↗
PubMed1992

The prevalence of carpal tunnel syndrome in patients with basal joint arthritis of the thumb.

Basal joint arthritis of the thumb and carpal tunnel syndrome are common conditions with an acknowledged coexistence. This article attempts to quantify the prevalence of carpal tunnel syndrome in patients with basal joint arthritis and to examine some of the etiologic factors that affect the coexistence of the two disorders. Of 246 patients who had surgery about the basal joint, 95 patients (39%) were identified by chart review as having carpal tunnel syndrome. Eleven of 122 remaining patients contacted had symptomatic carpal tunnel syndrome confirmed by nerve-conduction studies, bringing the total to 106 (43%). The prevalence was higher in worker's compensation patients and those with diabetes mellitus. The prevalence was lower in men than in women, and patients with inflammatory joint disease were at less risk than those with osteoarthritis. Given this high association, great care should be taken to diagnose or exclude coexistent carpal tunnel syndrome in patients scheduled for basal joint surgery so that, if present, it can be treated at the same time, diminishing the risk of postoperative morbidity and delayed symptoms.

Arthritis↗
PubMed1992

Inhibition of quantal transmitter release in the absence of calcium influx by a G protein-linked adenosine receptor at hippocampal synapses.

Spontaneous miniature excitatory postsynaptic currents (MEPSCs) were recorded by whole-cell voltage-clamp techniques in cultured rat hippocampal pyramidal neurons. The specific adenosine A1 receptor agonist cyclopentyladenosine (CPA) reduced the frequency of MEPSCs without affecting their amplitude distribution or kinetic properties. This action was blocked by pretreatment of the cells with pertussis toxin. In the presence of divalent cation Ca2+ channel blockers, CPA was still effective in reducing the frequency of MEPSCs. It was shown that this effect cannot be explained by changes in basal Ca2+ influx. These results suggest that neurotransmitters that produce presynaptic inhibition at hippocampal synapses utilize several mechanisms, one of which may involve inhibition of some component of the quantal release apparatus that occurs independently of inhibition of Ca2+ influx.

Adenosine↗