Is a rise in [CA]1 responsible for rapid changes in neuronal excitability, CA currents, extracellular ions and pH0 induced by brief anoxia?
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Calcium↗
Biomedical subjects
M E Morris
Publications and source records attributed to M E Morris.
Effect of an analgesic dose of aspirin on electrolyte disposition in healthy subjects.
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Adult↗
Competing pathways in drug metabolism. II. An identical, anterior enzymic distribution for 2- and 5-sulfoconjugation and a posterior localization for 5-glucuronidation of gentisamide in the rat liver.
Gentisamide (2,5-dihydroxybenzamide, GAM), a substrate that forms two monosulfates at the 2 and 5 positions (GAM-2S and GAM-5S) and a monoglucuronide at the 5 position (GAM-5G), was delivered at 8 or 80 microM by normal (N) and retrograde (R) flows to the once-through rat liver preparation. At the lower (8 microM) input concentration, ratios of conjugate formation rate, GAM-5S/GAM-5G and GAM-2S/GAM-5G, were decreased significantly (4.01 +/- 1.42 to 2.93 +/- 0.99, and 1.13 +/- 0.65 to 0.66 +/- 0.41, respectively) whereas a small but significant increase in the steady-state extraction ratio, E (0.89 +/- 0.029 to 0.94 +/- 0.016), was observed upon changing the flow direction from N to R. At the higher input GAM concentration (80 microM), conjugate formation rate ratios were relatively constant for GAM-5S/GAM-5G (1.18 +/- 0.08 to 1.11 +/- 0.12) and GAM-2S/GAM-5G (0.33 +/- 0.05 to 0.31 +/- 0.05) upon changing flow direction from N to R, despite a slight increase in E from 0.87 +/- 0.023 to 0.91 +/- 0.016 was observed. These results suggest that the sulfotransferase activities responsible for 2- and 5-sulfoconjugations are identically distributed and localized anterior to 5-glucuronidation activities during a normal flow of substrate into the rat liver (entering the portal vein and exiting the hepatic vein), and that the presence of uneven distribution of conjugation activities is discriminated only at the lower input drug concentration. At high concentration (greater than Km for all systems), saturation of all pathways occurs, and other anteriorly/identically distributed competing pathways would fail to perturb downstream intrahepatic drug concentrations and the resultant conjugation rates. The lack of change in metabolic profiles renders the condition unsuitable for examination of uneven distribution of enzymes in the liver. These observations were generally predicted by theoretical enzymic models of consistent distribution patterns. Because 2- and 5-sulfation were mediated by systems of similar Km but different Vmax values, two possibilities, the same isozyme of sulfotransferase being involved in the formation of two enzyme-substrate complexes to form two distinctly different products or two isozymes of sulfotransferases of identical distribution, were discussed.
Animals↗
Assay of inorganic sulfate in biologic fluids by nonsuppressed (single-column) ion chromatography.
An assay using nonsuppressed (single-column) anion chromatography was developed to determine the concentration of inorganic sulfate in biologic fluids. A conventional HPLC system with an anion-exchange column and conductimetric detector interfaced with an automatic injector and integrator was used. The mobile phase for the chromatography of urine and serum samples is 4 mM potassium hydrogen phthalate, pH 4.5, and potassium iodide is used as the internal standard. For cerebrospinal fluid samples, the mobile phase is modified by addition of 10% of a 4 mM phthalic acid solution. Results of the HPLC assay were found to correlate well (r = 0.991 and 0.999) with those of two commonly used spectrophotometric methods for urine and serum inorganic sulfate determinations. However, the concentrations determined by ion chromatography were 2.5 to 10% lower, possibly due to less assay interference by other substances following chromatographic separation of sulfate. Anion chromatography using a single-column system is a convenient and relatively inexpensive method with sufficient sensitivity for the determination of inorganic sulfate concentrations in urine, serum, and cerebrospinal fluid.
Animals↗
Sulfate homeostasis. I. Effect of salicylic acid and its metabolites on inorganic sulfate in rats.
Homeostasis of inorganic sulfate, a physiologic anion necessary for both detoxification and biosynthetic reactions, involves predominantly capacity-limited renal clearance mechanisms. The objective of this investigation was to examine the effect of salicylic acid (SA) and its major metabolites, salicyluric acid and salicyl phenolic glucuronide, on the serum concentrations and renal clearance of inorganic sulfate in rats. Animals were studied using a crossover design in which they received a bolus i.v. injection (75 mg/kg) and infusion (approximately 0.26 mg/min/kg) of SA or the same volume of saline (the vehicle). Blood samples were collected at 2, 3 and 4 hr after administration and urine between 2 and 4 hr. The renal clearance of sulfate and creatinine were examined at mean steady-state SA serum concentrations of 249 micrograms/ml. Although no changes in the serum concentrations and renal clearance of creatinine were observed, the renal clearance of inorganic sulfate was increased significantly (2.13 +/- 0.74 vs. 1.09 +/- 0.54 ml/min/kg in controls, mean +/- S.D., n = 7) and its serum concentration decreased (0.55 +/- 0.12 vs. 1.04 +/- 0.23 mM in controls). These changes were not due to alterations in uric acid concentrations as uric acid serum concentrations and renal clearance were unchanged when examined at similar steady-state SA serum concentrations in a subsequent study. The effects on sulfate disposition also were probably not due to the major metabolites of SA: no changes in the serum concentrations or renal clearance of sulfate were observed at mean steady-state concentrations of 52 micrograms/ml of salicyluric acid or 73.7 micrograms/ml of salicyl phenolic glucuronide after their direct administration.(ABSTRACT TRUNCATED AT 250 WORDS)
Animals↗
Competing pathways in drug metabolism. I. Effect of input concentration on the conjugation of gentisamide in the once-through in situ perfused rat liver preparation.
Sulfation and glucuronidation are two parallel pathways for the metabolism of phenolic substrates. Gentisamide (GAM) was used as a model compound to examine the effects of parallel competing pathways on drug disappearance and metabolite formation in the once-through perfused rat liver preparation. GAM was found to form one glucuronide (GAM-5G) and two sulfate (GAM-2S and GAM-5S) conjugates. These GAM conjugates were biosynthesized in recirculating rat liver preparations, and were isolated by preparative high-performance liquid chromatography. Specific incorporation of 35S-sodium sulfate and [14C]glucose into GAM sulfate and glucuronide conjugates revealed corresponding elution patterns as labeled GAM metabolites. Their identities were characterized by enzymatic and acid hydrolyses and by NMR spectroscopy. Gentisamide-5-sulfate (GAM-5S) and gentisamide-5-glucuronide (GAM-5G) are major metabolites, and gentisamide-2-sulfate (GAM-2S) is a minor metabolite. Single-pass rat liver perfusions were used to examine the effect of stepwise increases/decreases of input GAM concentration (CIn) on the extraction ratio (E) of GAM and formation of metabolites. The E of GAM remained constant (about 0.89) at input concentrations from 0.9 to 120 microM and decreased at CIn greater than 120 microM. Metabolite patterns, however, changed with GAM CIn, even when E was constant at CIn up to 120 microM. GAM-5S was present as the major metabolite of GAM at all GAM CInS in most liver preparations but the proportions of GAM-5S and GAM-2S decreased at increasing CIn; the proportion of GAM-5G, a minor metabolite at low CIn, increased with increasing CIn. Biliary excretion rates at steady state accounted for 5.3 +/- 2.7% (mean +/- S.D.) of the input rate: GAM-5G was the predominant metabolite found. Fitting the metabolic data (sum of the rates of efflux in bile and perfusate at steady state) and the logarithmic average drug concentration at the various CIn to the Michaelis-Menten equation furnished parameter estimates for the three metabolic pathways. The estimated Km and Vmax values were quite comparable: for GAM-2S formation, 22 microM and 287 nmol/min; for GAM-5S formation, 26 microM and 978 nmol/min; for GAM-5G formation, 71 microM and 1062 nmol/min, indicating that sulfation and glucuronidation are effective competing pathways of each other. In viewing the fate of GAM over the CIn range, a changing metabolic fate of GAM over a constant E was noted at CIn less than 120 microM: the sum of the rates of metabolite formation was proportional to CIn.(ABSTRACT TRUNCATED AT 400 WORDS)
Animals↗
Intracellular calcium in mammalian brain cells: fluorescence measurements with quin2.
Dispersed brain cells from 12-14 day old mouse embryos were loaded with the Ca2+-sensitive fluorescent probe, quin2 and shown to have a resting intracellular Ca2+ concentration ([Ca2+]i) of 158 nM (SE +/- 5) in the presence of 1 mM [Ca2+]o. When external [Ca2+] was raised from 0 to 1 mM there was an increase of [Ca2+]i of 70 nM; with further additions of Ca to greater than or equal to 10 mM [Ca2+]o the level of [Ca2+]i increased by less than or equal to 25 nM. Releasable intracellular Ca2+ stores, estimated from the increase in [Ca2+] produced by 4-Br A23187 in the absence of extracellular Ca2+, were 24 fmol/10(6) cells. A small increase in [Ca2+]i could be produced by the mitochondrial inhibitor, carbonyl cyanide m-chlorophenylhydrazone (CCCP). When extracellular K+ was raised by 10-20 mM, intracellular Ca2+ levels increased from 152 (SE +/- 7) to 204 nM (SE +/- 10). These K+-induced increases in [Ca2+]i were blocked by verapamil, did not occur in the absence of extracellular Ca2+, and presumably reflect the activation of voltage-dependent Ca2+ channels. N-methyl-D-aspartic acid (NMDA) evoked an increase in [Ca2+]i, while the kainate-like lathyrus sativus neurotoxin, L-3-oxalyl-amino-2-aminopropionic acid (L-3,2-OAP) did not; this is consistent with previous observations of different and respectively Ca2+-dependent and -independent mechanisms of action of these excitatory amino acids.
Amino Acids↗
Competition between two enzymes for substrate removal in liver: modulating effects due to substrate recruitment of hepatocyte activity.
Modulating effects of competing pathways, exemplified by sulfation (high affinity-low capacity) and glucuronidation (low affinity-high capacity), on drug disappearance and metabolite formation were investigated in a simulation study. The phenomenon of substrate recruitment of hepatocyte activity in drug removal and metabolite formation was shown with respect to inlet substrate concentration, and drug processing from inlet to outlet by enzyme systems localized differentially along the sinusoidal flow path in liver. Three enzymic distribution models: (A) sulfation and glucuronidation evenly distributed in liver, (b) sulfation occurring exclusively in the first half of the liver and glucuronidation in the second half, and (C) glucuronidation solely in the first half and sulfation in the second half, were described. The influence of Km and Vmax of the competing pathway, including enzyme induction (increase in Vmax), on any given pathway was also explored. Competing pathways exert their effects on other given pathways by modulating intrahepatic drug concentration from the inlet to outlet of the liver. When a competing pathway is similarly distributed or is at an anterior location to another pathway, the former pathway effectively reduces intrahepatic drug concentrations which reach downstream hepatocytes for recruitment of activity. For example, when glucuronidation activity is anterior to sulfation activity (defined with respect to flow direction), sulfation is without an effect on glucuronidation, but glucuronidation exerts a maximal influence over sulfation rates (Model C). When glucuronidation is in direct competition with sulfation (Model A) or is posteriorly distributed to sulfation (Model B), saturation of the high-affinity sulfation pathway leads to greater fluxes of substrate available downstream for glucuronidation. This results in an apparent compensatory increase in glucuronidation with reduced sulfation capacity, which occurs at input concentrations greater than the Km for sulfation but less than the Km for glucuronidation. This compensation pattern is more prominent for highly extracted compounds where both sulfation and glucuronidation are effective pathways in drug removal, and where large intrahepatic drug concentration gradients are expected. Since the physiologic description of intrahepatic drug concentration is often described by a concentration gradient from the inlet to outlet of the liver, the logarithmic average concentration has been used to estimate the mean liver concentration in the determination of kinetic constants for enzymic reactions.(ABSTRACT TRUNCATED AT 400 WORDS)
Glucuronidase↗
Intracellular calcium recordings from isolated cells of the mammalian central nervous system.
Measurements made with two different techniques of intracellular calcium levels from small isolated cells of the mammalian central nervous system are described and compared. Recordings in cultured mouse embryo spinal cord and dorsal root ganglion neurons, made with double-barrelled borosilicate Ca2+-selective microelectrodes yielded a mean Ca2+ level of 2.3 (SE +/- 0.54) microM for the lowest values recorded in 24 out of 46 cells. Intracellular Ca2+ dependence on membrane potential was apparent with levels of calcium greater than or equal to 4 microM (r = 0.371, n = 29). Both cyclic fluctuations induced by tetraethylammonium and an apparent increase in Ca2+ evoked by the depolarizing excitatory amino acid, L-aspartate, were observed. In contrast, estimates of intracellular Ca2+ obtained by spectrofluorimetry of suspensions of mouse embryo brain cells, loaded with the intracellular Ca-binding fluorescent probe, quin2 provided a approximately equal to 10-fold lower value, 152 (SE +/- 7) nM. This more closely resembles levels reported for large neurons where large-tip microelectrodes with greater sensitivity were used, and in spite of the heterogeneity of the cells this value is presumed to be a more accurate estimate of intraneuronal Ca2+ concentration. In these fluorescence studies KCl readily evoked increases in intracellular Ca2+ which could be blocked by verapamil and Cd2+ and were not induced in the absence of Ca2+. Increases were also produced by N-methyl-D-aspartate, but not by the kainate-like Lathyrus neurotoxin, L-3-oxalylamino-2-aminopropionic acid. These results provide preliminary evidence for both voltage-sensitive and receptor-activated Ca channels in embryonic brain cells.(ABSTRACT TRUNCATED AT 250 WORDS)
Animals↗
Measurements of intracellular free Ca2+ in mammalian central neurons.
Neutral carrier-containing Ca2+-selective microelectrodes were used to record the cytoplasmic free Ca2+ concentration [( Ca2+]i) in spinal cells in cats and in hippocampal cells of rats (in situ). The mean [Ca2+]i in motoneurons was close to 1 microM. Antidromic or direct stimulation for 30 s at 10 Hz increased [Ca2+]i by a mean of 90 nM. Such a small increase in [Ca2+]i and its slow decay (with a mean half-time of 23 (SD +/- 14.5) s) indicate very effective intracellular sequestration of Ca2+. Orthodromic stimulation consistently evoked smaller increases in [Ca2+]i. A much larger rise of interneuronal [Ca2+]i was evoked by stimulation of dorsal roots: by contrast intra-axonal recording (in motor or sensory fibres) failed to reveal any increase in [Ca2+]i in response to stimulation at 100 Hz. In the hippocampus, presumably because of poorer recording conditions, resting values of [Ca2+]i were higher (mean 8.5 microM). Repetitive stimulation of the fimbria--commissure at 5-20 Hz for 30 Hz, had variable effects on [Ca2+]i. Very large increases (to greater than 200 microM) were elicited repeatedly in some cells, either near the end of the tetanic stimulation or after a 20-30 s delay. Such major increases, which were associated with population cell discharges in bursts, may be related to long-term changes in hippocampal neuronal properties that are evoked by tetanic stimulation. Both in the spinal cord and the hippocampus, probable intraglial recordings showed relatively high mean levels of [Ca2+]i (about 30 microM).
Animals↗
Complex fractures of the proximal humerus: role of CT in treatment.
The authors reviewed the computed tomography (CT) scans, plain radiographs, and subsequent treatment of 17 patients with complex proximal humeral fractures. CT scans and radiographs were compared in the demonstration of fracture lines, displacement of fracture fragments, rotation of fragments relative to their normal positions, and status of the head and articular surface of the humerus. The impact of CT findings on the decision to treat with surgery versus closed reduction and on the choice of surgical procedure was assessed. Surgery was not performed in nine patients because CT scans showed no significant displacement of fragments previously judged displaced or "indeterminate" on radiographs. Surgery was performed in eight patients; CT demonstrated significant abnormalities not definitely shown with radiography. In six of these eight patients, CT scans demonstrated unsuspected abnormalities that directed the choice of surgical procedure. CT scans provide clinically useful information for the treatment of complex proximal humeral fractures when radiographs provide inadequate or indefinite information.
Adult↗
Absorption of magnesium from orally administered magnesium sulfate in man.
The use of magnesium sulfate (Epsom salt) as a cathartic in patients with impaired renal function can lead to severe toxicity due to hypermagnesemia. Although toxicity is uncommon in healthy subjects, little is known concerning the extent of absorption of magnesium after a cathartic dose of magnesium sulfate. The bioavailability of magnesium following a large oral dose of magnesium sulfate in normal volunteers was examined in the present investigation. Baseline 24-hour urinary excretion rates of magnesium and creatinine were determined over 3 consecutive days in 6 healthy men. The oral administration of 13.9 g (56.5 mmoles) magnesium sulfate U.S.P., in 4 equal hourly increments, resulted in the urinary excretion (corrected for baseline excretion rate) of 4.0 +/- 2.9% (mean +/- SD) of the dose of magnesium during the first 24 hours and 6.9 +/- 7.0% of the dose during a 72-hour interval. Magnesium sulfate administration had no effect on the 24-hour urinary excretion rate of creatinine. The baseline excretion rate of magnesium was significantly correlated with that of creatinine (r = 0.875) and inorganic sulfate (r = 0.921). All of the subjects experienced mild or moderate diarrhea. Therefore, magnesium is absorbed to a limited and variable extent in healthy adults following a cathartic dose of magnesium sulfate.
Administration, Oral↗
Changes in free calcium ion concentration recorded inside hippocampal pyramidal cells in situ.
In rats under urethane or pentobarbitone anesthesia, Ca2+ -sensitive microelectrodes were inserted into CA3 and CA1 hippocampal cells. In 23 neurons with a mean resting membrane potential (Vm) of -56.9 mV, the Ca potential (VCa) fell below Vm by an average of -22.1 mV (S.D. +/- 19.1 mV), indicating a mean intracellular free Ca2+ concentration ([Ca]i) of 9.7 microM (S.D. 14.9 microM). In spite of their better and more stable Vm (mean -67.1 mV), unresponsive cells (probably neuroglia) had a higher and more variable [Ca]i (mean 37.0 +/- 51.2 microM). In 21 of the neurons, repetitive stimulation of the fimbria--at 5-20 Hz for 30s, which is sufficient to elicit bursts of population spikes--evoked substantial increases in [Ca]i: the mean increase observed during or just after 29 such tetani was +27.1 +/- 54.5 microM. Typically [Ca]i reached a peak near the end of the tetanus and then decayed with a half-time of 5-10 s, though not necessarily to the initial level. In 7 cells, a large increase in [Ca] (mean +239 +/- 367 microM) appeared as a late event, 20-30 s after the end of the tetanus. In 5 cells, [Ca]i could thus be raised transiently to 10(-4) M or higher. All these increases in [Ca]i are far greater than can be evoked by tetanic activation in spinal motoneurons; their possible significance for long term potentiation or cell necrosis in the hippocampus is discussed.
Animals↗
Effect of acetaminophen on inorganic sulfate concentrations in human cerebrospinal fluid.
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Acetaminophen↗
Study on the application of the light-cured composite resin inlay in primary molars (I).
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Child↗
Improved survival of the diabetic foot: the role of a specialized foot clinic.
A specialized foot clinic for diabetic patients has made a detailed analysis of the presentation of diabetic foot ulcers and from this a new, organised approach to treatment has been derived. Over three years it has achieved a high rate of ulcer healing and reduced the number of major amputations. It has brought together the skills of chiropodist, shoe-fitter, nurse, physician and surgeon to manage the distinctive lesions of the neuropathic and ischaemic diabetic foot. The neuropathic ulcer was invariably associated with callus, whereas the ischaemic ulcer presented as areas of necrosis often from localised pressure of tight shoes. Essential aspects of management are specially constructed shoes, intensive chiropody and precise antibiotic treatment. Healing was achieved in 204 out of 238 (86 per cent) neuropathic ulcers and 107 out of 148 (72 per cent) ischaemic ulcers. Relapse rate in special shoes was 26 per cent compared with 83 per cent who preferred to wear their own shoes. In the two years before the establishment of the clinic, there were 11 and 12 major amputations yearly. This rate has now been reduced to seven, seven and five amputations yearly.
Aged↗
Identification of extrasynaptic binding sites for [3H]GABA in peripheral nerve.
The binding of radiolabelled gamma-aminobutyric acid ( [3H]GABA) to extrasynaptic sites in peripheral nerve was examined in order to characterize the receptor species mediating the depolarizing action of GABA on myelinated axons. Binding of [3H]GABA was carried out under Na+-free conditions with fresh homogenates of amphibian sciatic nerve. The kinetics of association of the radioligand suggested the presence of a rapidly associating, reversible binding site, and a slowly associating, apparently irreversible one. Scatchard plots of the reversible component of binding were linear and yielded a mean affinity (Kd) of 22 nM. This stands in contrast with the ED50 of 90 microM for GABA-evoked depolarization of frog nerve; and therefore the high affinity binding sites are unlikely to be involved in such depolarization. The rank order of potency of agonists that competed with [3H]GABA was muscimol greater than GABA much greater than delta-aminovaleric acid greater than beta-alanine. However, the GABAA analogues 3-aminopropanesulfonic acid, beta-guanidinopropionic acid, and guanidoacetic acid as well as the GABAB agonist baclofen all failed to displace the ligand. Bicuculline methiodide, picrotoxin, and nipecotic acid also did not compete. The bicuculline-insensitive, baclofen-insensitive high affinity binding sites identified here appear to be unique, as they are distinctly different from the classical GABAA and GABAB receptors.
Animals↗
Pharmacodynamics of the hypnotic effect of salicylamide in rats.
Salicylamide (SAM) can produce sedation and sleep in humans and animals. To explore the potential utility of the drug as a research tool for assessing disease effects on the response of the central nervous system to depressant drugs, and to obtain a better understanding of the clinically evident sedative action of SAM, studies were performed to characterize the relationship between the concentrations and hypnotic effect of this drug in rats. Female Lewis rats weighing 170-200 g received SAM by intravenous infusion at a rate of 0.49, 1.22, or 2.47 mg/min until the onset of loss of the righting reflex. This well-defined pharmacological endpoint occurred from 16.7 +/- 2.3 min (fastest infusion rate) to 110 +/- 27 min (slowest infusion rate) after the start of the infusion. SAM concentrations at that time in serum, serum water, brain, and cerebrospinal fluid (CSF) were similar in animals that had received the 1.22- or 2.47-mg/min infusion and lower in animals that were infused at a rate of 0.49 mg/min. The slowest infusion rate group also exhibited increased serum protein binding of the drug. The SAM concentration ratio, CSF-serum water, was essentially unity in all three groups, indicative of rapid equilibration of the drug across the blood-CSF barrier. Gentisamide, the hydroxylated metabolite of SAM, was found in serum, CSF, and brain, but in low concentrations at which this metabolite alone had no hypnotic effect.(ABSTRACT TRUNCATED AT 250 WORDS)
Animals↗