Biomedical subjects

G W Miller

Publications and source records attributed to G W Miller.

At least 37 records · Page 2Linked to original sources

PubMed1997

Radioligand binding and immunoautoradiographic evidence for a lack of toxicity to dopaminergic nerve terminals in human cocaine overdose victims.

Radioligand binding to and immunolabeling of transport sites associated with monoamine-containing synaptic vesicles affords a novel approach for mapping the integrity of dopaminergic (DAergic) nerve terminals. The present study used [125I]iodovinyltetrabenazine ([125I]TBZ) and a fusion protein antibody directed at the large intraluminal loop of the neuronal vesicular monoamine transporter (hVMAT2-loop) as probes to assess the effects of chronic cocaine use on the integrity of DAergic nerve terminals in the striatum of cocaine fatalities. Visualization of [125I]TBZ binding in human brain revealed a distinct pattern of labeling throughout the rostral-caudal extent of the striatum. Saturation binding of [125I]TBZ in striatal membranes demonstrated a single high affinity site (Kd = 2.3 +/- 0.9 nM and Bmax = 55.5 +/- 8.1 pmol/g tissue) with a pharmacological profile (tetrabenazine > or = iodovinyltetrabenazine > ketanserin > or = reserpine > haloperidol > GBR 12909) consistent with the specific labeling of hVMAT2. Quantitative in vitro autoradiography demonstrated no significant alteration in the density of [125I]TBZ binding sites in the anterior and posterior sectors of the striatum in cocaine fatalities with and without preterminal excited delirium as compared to drug-free and age-matched control subjects. Similarly, the levels of hVMAT2-loop immunoreactivity were not significantly different across control and cocaine fatality groups. The results demonstrate the lack of an alteration in [125I]TBZ binding sites and hVMAT2 protein in the striatum from a young cohort of cocaine fatalities. Since striatal VMAT2 is primarily associated with DAergic nerve terminals, these results suggest that chronic cocaine use failed to affect the integrity of striatal DAergic nerve terminals.

Adolescent↗

PubMed1997

Immunochemical analysis of dopamine transporter protein in Parkinson's disease.

The plasma membrane dopamine transporter (DAT) is considered to be a reliable marker of presynaptic dopaminergic terminal loss. Previous in vivo imaging and postmortem binding studies have detected a loss in striatal DAT binding in Parkinson's diseased (PD) brain; however, these techniques have poor spatial resolution and may suffer from nonspecific binding of some ligands. In this study, we use novel highly specific monoclonal antibodies to distinct epitopes of human DAT to quantify and localize the protein. Western blot analysis revealed marked reductions in DAT immunoreactivity in putamen, caudate, and nucleus accumbens of PD brain compared with control cases, and the reductions were significantly correlated to disease duration. Immunohistochemistry revealed DAT-immunoreactive fibers and puncta that were dense throughout the striatum of control brains but that were drastically reduced in putamen of PD brains. Caudate from PD brains showed a significant degree of sparing along the border of the ventricle, and the nucleus accumbens was relatively preserved. An unexpected finding was that discrete islands of DAT immunoreactivity were preserved within the matrix of PD putamen. Thus, immunological analysis of DAT protein provides novel and sensitive means for localizing and quantifying DAT protein in PD and other neurological disorders involving dopaminergic systems.

Aged↗

PubMed1997

Knockout of the vesicular monoamine transporter 2 gene results in neonatal death and supersensitivity to cocaine and amphetamine.

Vesicular monoamine transporters are known to transport monoamines from the cytoplasm into secretory vesicles. We have used homologous recombination to generate mutant mice lacking the vesicular monoamine transporter 2 (VMAT2), the predominant form expressed in the brain. Newborn homozygotes die within a few days after birth, manifesting severely impaired monoamine storage and vesicular release. In heterozygous adult mice, extracellular striatal dopamine levels, as well as K+- and amphetamine-evoked dopamine release, are diminished. The observed changes in presynaptic homeostasis are accompanied by a pronounced supersensitivity of the mice to the locomotor effects of the dopamine agonist apomorphine, the psychostimulants cocaine and amphetamine, and ethanol. Importantly, VMAT2 heterozygous mice do not develop further sensitization to repeated cocaine administration. These observations stress the importance of VMAT2 in the maintenance of presynaptic function and suggest that these mice may provide an animal model for delineating the mechanisms of vesicular release, monoamine function, and postsynaptic sensitization associated with drug abuse.

Amphetamine↗

PubMed1997

Neurosteroid inhibition of cell death.

Diverse gamma-aminobutyric acid (GABAA) receptor modulators exhibited novel cytoprotective effects and mechanisms of action in rabbit renal proximal tubules subjected to mitochondrial inhibition (antimycin A) or hypoxia. Cytoprotective potencies (50% effective concentration, EC50) were 0.3 nM allopregnanolone (AP) > 0.4 nM 17 alpha-OH-allopregnanolone (17 alpha-OH-AP) > 30 nM dehydroepiandrosterone sulfate (DHEAS) = 30 nM pregnenolone sulfate (PS) > 500 nM pregnenolone (PREG) > 30 microM muscimol > 10 mM GABA following antimycin A exposure. Maximal protection with AP and 17 alpha-OH-AP was 70%, whereas DHEAS, PS, PREG, and muscimol produced 100% cytoprotection. Experiments with AP, PS, and muscimol revealed the return of mitochondrial function and active Na+ transport following hypoxia/reoxygenation. Muscimol inhibited the antimycin A-induced influx of both extracellular Ca2+ and Cl- that occurs during the late phase of cell injury, whereas the neurosteroids only inhibited influx of Cl-. Radioligand binding studies with AP and PS did not reveal a specific binding site; however, structural requirements were observed for cytoprotective potency and efficacy. In conclusion, we suggest that the GABAA receptor modulators muscimol and neurosteroids are cytoprotective at different cellular sites in the late phase of cell injury; muscimol inhibits Ca2+ and subsequent Cl- influx, whereas the neurosteroids inhibit Cl- influx.

Alkaloids↗

PubMed1995

Involvement of an axonal reflex in IgE-mediated inflammation in mouse skin.

A neurogenic component of IgE-mediated inflammation was demonstrated in mice by footpad denervation. Footpad swelling was reduced 26% following sciatic nerve transection, but unaffected by rhizotomy or spinal nerve transection. These data provide in vivo evidence that an axonal reflex is involved in IgE-mediated inflammation and completed distal to the cell bodies of the sensory neurons located in the lumbar spinal ganglia. Furthermore, depletion of neuropeptides with capsaicin also reduced IgE-mediated swelling by 26%, indicating that unmyelinated axons are involved in the neurogenic component of IgE-mediated inflammation.

Animals↗

PubMed1995

Inhibitors of renal chloride transport do not block toxicant-induced chloride influx in the proximal tubule.

We have previously demonstrated that chloride influx occurs during the late stages of mitochondrial inhibitor-induced renal proximal tubule (RPT) cell injury. The purpose of this study was to determine if chloride influx is a common pathway in toxicant-induced cell injury and if inhibitors of renal chloride transport block the chloride influx. Chloride influx occurred in the late stages of RPT cell injury induced by the diverse toxicants mercuric chloride, t-butyl hydroperoxide, bromohydroquinone, and tetrafluoroethyl-L-cysteine. Specific inhibitors of known renal chloride transport did not prevent antimycin A-induced chloride influx. Toxicant-induced chloride influx occurred prior to cell swelling and decreasing the extracellular chloride concentration diminished toxicant-induced cell death. Thus, chloride influx is a common pathway in the late stages of toxic cell injury and does not occur through known mechanisms of renal chloride transport. Further, we propose that toxicant-induced chloride influx is mediated by a novel receptor related to the neuronal strychnine-sensitive glycine receptor and that chloride influx is a key step in cell swelling and lysis.

Animals↗

PubMed1995

Immunocytochemical detection of tumor cells in bone marrow and peripheral blood stem cell collections from patients with ovarian cancer.

High-dose chemotherapy (HDC) followed by autologous hematopoietic reconstitution is an experimental treatment option for patients with epithelial ovarian cancer. However, the incidence of occult ovarian tumor cell involvement in autologous bone marrow (BM) or peripheral blood stem cell (PBSC) autografts has not been widely investigated. We used a highly sensitive immunocytochemical (ICC) procedure that detects occult blood-borne tumor micrometastases. We analyzed 24 BM specimens (15 obtained during therapy and 9 harvest samples) and seven PBSC specimens from 22 patients with ovarian cancer. Overall, ICC analysis detected immunostained tumor cells in 10 of 23 evaluable BM specimens (43%) from 9 of 19 patients (47%). One of 9 (11%) harvest samples contained tumor cells. Only one of the 10 ICC-positive BM specimens had tumor cells detected by routine histopathological analysis. ICC-detectable tumor cells were cleared from the marrow of two patients during chemotherapy. None of the seven PBSC specimens contained tumor cells. We conclude that ovarian cancer micrometastases have the potential to contaminate BM, as is also the case in patients with other epithelial malignancies. In the limited number of specimens analyzed, PBSC harvests appeared to provide a less tumor-contaminated source of hematopoietic stem cells for autologous transplantation.

Antibodies, Monoclonal↗

PubMed1994

Strychnine and glycine protect renal proximal tubules from various nephrotoxicants and act in the late phase of necrotic cell injury.

We have previously shown that strychnine mimics the cytoprotective properties of glycine in renal proximal tubule (RPT) suspensions exposed to antimycin A (AA). The aims of this study were to determine whether the cytoprotective properties of strychnine applied to various types of nephrotoxicants and to examine the temporal aspects of the cytoprotection of glycine and strychnine. Tubular release of LDH activity was used as a marker of cell death. Glycine (2 mM) or strychnine (1 mM) added 5 min prior to the toxicant decreased LDH release in rabbit RPT suspensions exposed to 25 microM tetrafluoroethyl-L-cysteine (TFEC), 10 microM HgCl2, 0.5 mM t-butyl hydroperoxide (TBHP), or 0.2 mM bromohydroquinone (BHQ) for 4 hr, or 2 mM sodium cyanide (NaCN) for 2 hr. The relative rank order of effectiveness of glycine and strychnine was NaCN = TFEC > BHQ > DCVC >> TBHP > HgCl2. The temporal aspects of strychnine and glycine protection were examined by exposing RPT to either AA or TFEC for 1 or 3 hr, respectively, and then adding either 1 mM glycine or 1 mM strychnine. Glycine and strychnine decreased LDH release in AA-treated RPT at 1.25 and 2 hr and TFEC-treated RPT at 4 hr. In addition, when RPT exposed to AA or TFEC and treated with strychnine or glycine were washed at either 1 or 4 hr, protection was eliminated at later time points. When glycine was added to RPT treated with either PCBC, TFEC, or DCVC 5 min prior to or 30, 60, 120, and 180 min following toxicant addition, LDH release was reduced at all time points. These results demonstrate that strychnine and glycine protect RPT from a variety of diverse nephrotoxicants, strychnine and glycine do not need to be present at the time of toxic insult, strychnine and glycine cytoprotection is reversible, and strychnine and glycine act in the late phase of necrotic cell injury.

Animals↗

PubMed1994

A putative cytoprotective receptor in the kidney: relation to the neuronal strychnine-sensitive glycine receptor.

The neutral amino acid glycine has been demonstrated to prevent cell death in numerous cell types exposed to a variety of toxic insults. Recently, the central nervous system (CNS) glycine antagonist strychnine was demonstrated to bind specifically to the plasma membrane of renal proximal tubules (RPT) and mimic glycine cytoprotection. Further, it has been demonstrated in RPT that glycine and strychnine block chloride influx in the late stages of cell injury. The aim of this study was to determine if the RPT cytoprotective site is related to neuronal glycine receptors. Only antagonists to the CNS strychnine-sensitive glycine receptor (strychnine, brucine), and not antagonists to the glycine modulatory site of the NMDA receptor (DCQX, 7-CKA, HA-966) or the GABAA receptor (bicuculline methiodide, picrotoxin), prevented mitochondrial inhibitor (antimycin A)-induced RPT cell death. Using immunoblot analysis, proteins corresponding to the 58 kDa beta-subunit of the strychnine-sensitive glycine receptor and the associated protein gephyrin were identified in rabbit kidney cortical membrane fractions and RPT. No protein corresponding to the 48 kDa alpha-subunit was identified. Thus, glycine and strychnine may exert their cytoprotective effects via a putative plasma membrane receptor that is related to the strychnine-sensitive glycine receptor found in the CNS.

Animals↗

PubMed1993

A novel low-affinity strychnine binding site on renal proximal tubules: role in toxic cell death.

Previous studies have shown that the neuronal glycine receptor antagonist, strychnine, mimics the cytoprotective effects of glycine in renal proximal tubules (RPT) (1). The goal of this study was to identify and characterize the site of action of strychnine. 3H-Strychnine bound to RPT in a saturable and reversible manner, and was displaced by unlabelled strychnine (IC50 = 0.87 mM and a Bmax = 57 nmol/mg protein). However, strychnine binding was not inhibited by glycine or related cytoprotective amino acids. Furthermore, the neurotoxicants bicuculline and norharmane, which share the cytoprotective properties of strychnine, inhibited 3H-strychnine binding. These data support the existence of a novel low-affinity strychnine binding site on the RPT plasma membrane that prevents toxic cell death.

Animals↗

PubMed1993

Cytoprotection by inhibition of chloride channels: the mechanism of action of glycine and strychnine.

Previous studies have demonstrated that strychnine mimics the cytoprotective effects of glycine (1) and that strychnine binds specifically to renal proximal tubules (RPT) at cytoprotective concentrations (2). The goal of this study was to determine a mechanism by which strychnine and glycine are cytoprotective. Antimycin A (0.1 microM) caused chloride influx subsequent to mitochondrial inhibition and prior to the release of lactate dehydrogenase (LDH) activity (a marker of cell death/lysis). The addition of strychnine or glycine prevented the chloride influx and LDH release. The chloride channel inhibitors ethacrynic acid, furosemide, anthracene-9-carboxylic acid, DIDS, and SITS decreased LDH release in RPT exposed to antimycin A with a rank order of potency of DIDS > ethacrynic acid = furosemide = anthracene-9-carboxylic acid > SITS. These data, in conjunction with the preceeding paper, indicate a critical role for chloride influx in cell death/lysis; support the existence of a novel strychnine binding site on the plasma membrane of RPT that is coupled to a chloride channel; and suggest that glycine and strychnine are cytoprotective through their inhibition of chloride influx.

Animals↗

PubMed1993

On the left and right hemisphere visual processing that precedes recognition.

Three experiments, and a replication of each, investigated the nonconscious prerecognition visual processing given left visual field (LVF) and right visual field (RVF) letter inputs. Each input was a vertically arrayed pair of letters in which three variables were manipulated: (1) the same letter twice vs one each of two letters, (2) same vs 180 degree difference in orientations within each letter pair, and (3) normal vs mirror-image letter form. The procedure presented all pairs of letter combinations in pairs of pre- and postmasked 10-msec flashes; the subject's task was to report which flash of each pair appeared to last longer. When letter pairs differed on all three variables, RVF presentations of mirror-image letters were judged to be longer than equal presentations of normal letters; the reverse occurred for LVF presentations. When one normal and mirror-image letter were presented, RVF presentations of mismatched orientations were judged to be longer than matched orientations, and the reverse was true for LVF presentations. When pairs of two normal letters were presented, no processing difference between LVF and RVF presentations was observed. A fourth experiment tested presence/absence detection of the letter pairs under the input conditions of the main experiments and showed those conditions to produce chance-level presence/absence detection. These results suggest that each hemisphere can perform its own prerecognition operations and that neither hemisphere is necessarily specialized for any particular prerecognition visual operation.

Adult↗

PubMed1992

Evidence of a neurogenic component during IgE-mediated inflammation in mouse skin.

IgE-mediated inflammation was measured in mouse footpads that lacked sciatic innervation. Mice were passively sensitized with a monoclonal antibody, IgE anti-dinitrophenol, or were immunized for specific IgE production. Antigen-induced swelling in the denervated footpads was reduced 23-39% when compared to sham or untreated controls. Reduced IgE-mediated swelling responses were attributed to the loss of a mast cell-nerve interaction and not to blood vessel sensitivity to vasoamines. Furthermore, electrical stimulation of the distal segment of the sciatic nerve completely restored IgE-mediated inflammation. These data provide in vivo evidence that peripheral nerves participate in cutaneous IgE-mediated swelling reactions with the net effect of increasing inflammation.

Analysis of Variance↗

PubMed1992

Effects of phosphate loading on metabolic and myocardial responses to maximal and endurance exercise.

Six trained male cyclists and triathletes participated in a double blind study to determine the effects of phosphate loading on maximal and endurance exercise performance. Subjects ingested either 1 gm of tribasic sodium phosphate or a glucose placebo four times daily for 3 days prior to performing either an incremental maximal cycling test or a simulated 40-km time trial on a computerized race simulator. They continued the supplementation protocol for an additional day and then performed the remaining maximal or performance exercise test. Subjects observed a 17-day washout period between testing sessions and repeated the experiment with the alternate supplement regimen in identical fashion. Metabolic data were collected at 15-sec intervals while venous blood samples and 2D-echocardiographic data were collected during each stage of exercise during the maximal exercise test and at 8-km intervals during the 40-km time trial. Results indicate that phosphate loading attenuated anaerobic threshold, increased myocardial ejection fraction and fractional shortening, increased maximal oxidative capacity, and enhanced endurance performance in competitive cyclists and triathletes.

Adult↗

PubMed1990

Effects of phosphate loading on oxygen uptake, ventilatory anaerobic threshold, and run performance.

Seven male competitive runners (VO2max 73.9 +/- 6.3 ml.kg-1.min-1) participated in a two-session, placebo, double-blind study to determine the effects of phosphate loading on oxygen uptake, ventilatory anaerobic threshold, and 5-mile run performance. Subjects ingested 1000 mg of tribasic sodium phosphate or a placebo four times daily for 6 d. A maximal running stress test or a 5-mile performance run was performed randomly on either the 3rd or the 6th d. Test sessions were separated by a 2-wk washout period and repeated with alternating phosphate and placebo regimens. Venous blood samples were collected prior to and following each max and run session. Results revealed that placebo resting serum phosphate levels were mildly elevated and that phosphate loading significantly increased resting and post-exercise serum phosphate values. Resting and post-exercise 2,3-diphosphoglycerate values were decreased while hemoglobin values were elevated with phosphate ingestion. Phosphate loading significantly increased maximal oxygen uptake from 4.77 +/- 0.29 to 5.18 +/- 0.25 l.min-1 and ventilatory anaerobic threshold from 3.74 +/- 0.28 to 4.18 +/- 0.14 l.min-1. Five-mile run times were nonsignificantly different between placebo and phosphate sessions. However, mean performance run oxygen uptake was significantly lower (3.87 +/- 0.3 to 3.80 +/- 0.3 l.min-1) with phosphate ingestion. Data demonstrate that maximal and run performance were influenced by elevations in serum phosphate eliciting an increased maximal oxygen uptake, ventilatory anaerobic threshold, and variable effects on 5-mile run performance. These adaptations occurred without observable increases in red cell 2,3-diphosphoglycerate.

Anaerobic Threshold↗

PubMed1988

Pneumolabyrinth as a late sequela of temporal bone fracture.

Pneumolabyrinth, air in the labyrinthine system, is an uncommon entity. We report one case, adding to previous reports of two such cases. The diagnosis was made radiologically, which has become possible only recently with the advent of high-resolution computed tomography (CT) scanners; we suspect it will be used more often in the future. To our knowledge, this is the first case in which pneumolabyrinth has been identified as a delayed sequela of a temporal bone fracture. Furthermore, this is the first description of a pneumolabyrinth occurring in a transverse temporal bone fracture.

Humans↗

PubMed1987

Cochlear implant: audiometric results with a single-channel device.

With the FDA approval of the first cochlear implant device in November 1984, the possibility of the perception of sound became a reality for the profoundly deaf individual. The cochlear implant group of Cincinnati was involved in the coinvestigator study before the approval of the device. This paper presents data from seven patients with cochlear implants and the resulting improvement in their hearing perception, as well as a discussion about which patients can benefit from the cochlear implant.

Adult↗

PubMed1987

Membrane Transport in Isolated Vesicles from Sugarbeet Taproot: III. Effects of Fluoride on ATPase Activity and Transport.

The effects of fluoride on the tonoplast type ATPase and transport activities associated with sealed membrane vesicles isolated from sugarbeet (Beta vulgaris L.) storage tissue were examined. This anion had two distinct effects upon the proton-pumping vesicles. When ATP hydrolysis was measured in the presence of gramicidin D, significant inhibition (approximately 50%) only occurred when the fluoride concentration approached 50 millimolar. In contrast, the same degree of inhibition of proton transport occurred when the fluoride concentration was about 24 millimolar. Effects on proton pumping at this concentration of fluoride could be attributed to an inhibition of chloride movement which serves to dissipate the vesicle membrane potential. Valinomycin could partially restore ATPase activity in sealed vesicles which were inhibited by fluoride and this restoration occurred with a reduction in the membrane potential. Fluoride demonstrated a competitive interaction with chloride-stimulation of proton transport and inhibited the uptake of radioactive chloride into sealed vesicles. When the vesicles were allowed to develop a pH gradient in the absence of KCl, and KCl was subsequently added, fluoride reduced enhancement of the existing pH gradient by KCl. The results are consistent with a chloride carrier that is inhibited by fluoride.

Journal Article↗