Search PubMed⌕ Search

Biomedical subjects

A Ames

Publications and source records attributed to A Ames.

At least 19 recordsLinked to original sources
PubMed2000

CNS energy metabolism as related to function.

Large amounts of energy are required to maintain the signaling activities of CNS cells. Because of the fine-grained heterogeneity of brain and the rapid changes in energy demand, it has been difficult to monitor rates of energy generation and consumption at the cellular level and even more difficult at the subcellular level. Mechanisms to facilitate energy transfer within cells include the juxtaposition of sites of generation with sites of consumption and the transfer of approximately P by the creatine kinase/creatine phosphate and the adenylate kinase systems. There is evidence that glycolysis is separated from oxidative metabolism at some sites with lactate becoming an important substrate. Carbonic anhydrase may play a role in buffering activity-induced increases in lactic acid. Relatively little energy is used for 'vegetative' processes. The great majority is used for signaling processes, particularly Na(+) transport. The brain has very small energy reserves, and the margin of safety between the energy that can be generated and the energy required for maximum activity is also small. It seems probable that the supply of energy may impose a limit on the activity of a neuron under normal conditions. A number of mechanisms have evolved to reduce activity when energy levels are diminished.

Adenosine Triphosphate↗
PubMed1999

An in vitro rabbit retina model to study electrophysiologic and metabolic function during and following ischemia.

Most in vitro studies involving neuronal ischemia use biochemical measures and/or cell counting to assess cellular death. We describe an in vitro rabbit retina model in which we measured glucose utilization, lactate production, and light-evoked compound action potentials (CAPs) to assess metabolic and functional recovery following ischemia. Under control conditions, retinal glucose utilization and lactate production (n = 7), as well as CAPs (n = 8) remained quite constant for 6-8 h. During ischemia (glucose reduced from 6 to 1 mM and oxygen from 95 to 15%), glucose utilization and lactate production fell to 50%. CAPs fell to 50% in 3-4 min, and to 0% in 8-10 min. Recovery during 3-4 h of 'return-to-control' was dependent upon the length of ischemia. Glucose utilization recovered to 63% after 1 h (n = 4) and to 18% after 2 h of ischemia (n = 6, P < 0.001). Lactate production recovered to 77% after 1 h (n = 4) and to 54% after 2 h of ischemia (n = 6, P < 0.001). CAPs returned to 51, 15, and 0.13% of the control responses after 0.5 h (n = 7), 1 h (n = 8), and 2 h (n = 5) of ischemia, respectively (P < 0.001). This avascular, blood-brain barrier-free preparation provides an opportunity to use both metabolic and functional criteria to test protection against neuronal ischemia.

Action Potentials↗
PubMed1998

Work transitions.

The healthcare environment is changing rapidly. Work reorganization and redesign can lead to department and unit closures, resulting in work transitions for many staff members. The authors describe the findings of focus groups held with staff and management on units that have closed. These interviews highlight the difficulties encountered with change and provide suggestions to reduce uncertainty during department or unit closures.

Academic Medical Centers↗
PubMed1998

Induced hypertension improves regional blood flow and protects against infarction during focal ischemia: time course of changes in blood flow measured by laser Doppler imaging.

OBJECTIVE: To characterize changes in regional blood flow (rCBF) during and after a period of arterial occlusion and determine the effect on rCBF and on the extent of infarction when the mean arterial blood pressure is increased during the period of occlusion. METHODS: rCBF in the middle cerebral artery (MCA) territory of rabbits was monitored using laser Doppler perfusion imaging before, during, and after a 1- or 2-hour period of MCA occlusion, and the size of the infarction was assessed by 2,3,5-triphenyltetrazolamine chloride staining after 2 hours of reperfusion. Test animals, the mean arterial blood pressure of which was increased by 65 mm Hg with intravenous phenylephrine during the ischemia, were compared with control animals that remained normotensive. The laser Doppler perfusion imager (Lisca Developments Co., Linköping, Sweden) scanned a 3-cm2 area of cortex with a resolution of 4 mm2 every 15 minutes. RESULTS: MCA occlusion reduced rCBF to 71 +/- 2% of the control level (n = 24, P < 0.001). Hypertension (HTN) restored rCBF to 84 +/- 3% of the control level (n = 12, P < 0.01), but the HTN-induced improvement diminished with time, so that after 1 hour, there was no longer a significant difference between hypertensive and normotensive animals. HTN during the MCA occlusion caused a 97% reduction in infarct size (P < 0.05) in the animals subjected to 1 hour of occlusion but caused only a 45% reduction (P approximately 0.1) in the animals subjected to 2 hours of occlusion. CONCLUSION: This study supports the use of HTN to minimize ischemic injury from short intervals of major intracranial vessel occlusion but fails to demonstrate protection when HTN is maintained during occlusions of more than 1 hour.

Animals↗
PubMed1995

Protection against CNS ischemia by temporary interruption of function-related processes of neurons.

Previous studies have shown that most of the energy consumption of CNS tissue is used for processes that subserve signaling functions of the cells. Since these function-related processes are probably not essential to cell viability, blocking them reversibly with a combination of pharmacologic agents should protect cells from a reduction in energy metabolism. Preliminary experiments to test this hypothesis were performed on isolated rabbit retinas. They were maintained in a newly devised chamber that permitted continuous monitoring of electrophysiological function for > or = 8 h. Ischemia was simulated by a 6-fold reduction in both O2 and glucose. This caused a rapid (t1/2 75 s) and complete loss of the light-evoked response in the optic nerve. Untreated retinas showed full recovery after 1/2 h of deprivation, but only 50% recovery after 1 h and little or no recovery after 2 or 3 h. Retinas exposed during 3 h of deprivation to a combination of six agents that abolished electrophysiologic function and reduced glucose utilization [tetrodotoxin (TTX), 2-amino-4-phosphonobutyric acid (APB), 2-amino-5-phosphonovaleric acid (APV), amiloride, Mg2+, and Li+] showed full recovery. We conclude that reducing energy requirements by blocking functional processes can prevent ischemic damage.

Animals↗
PubMed1994

Steady state feedback in mammalian phototransduction illustrated by a nomogram.

Published data characterizing the four reactions responsible for the Ca(2+)-mediated negative feedback in mammalian rod phototransduction were used to generate graphs which are combined in a circular fashion so that the y-axis of one serves as the x-axis of the next. The nomogram thus created makes it possible to determine by inspection the steady state situation in darkness, and the quasi-steady state situations that pertain shortly after exposure to light of different intensities. The results predicted by the nomogram suggest that Ca(2+)-mediated negative feedback is responsible for the Weber-Fechner relationship between stimulus and response.

1-Methyl-3-isobutylxanthine↗
PubMed1992

Assessing work retention issues.

Within 6 months, a 10-member Nursing Services task force created and administered a literature-based retention survey. Tool development and administration is described. The 33-item tool was completed by 85% of the staff. Responses led to the formation of work groups, charged with addressing dissatisfiers and communicating implementation strategies to the hospital at large.

Adult↗
PubMed1992

Energy requirements of CNS cells as related to their function and to their vulnerability to ischemia: a commentary based on studies on retina.

CNS tissue is well known to have large energy requirements. However, because of the difficulty of measuring rates of energy usage, relatively little is known about which cell types and which neurophysiological functions are the principal energy users. In experiments performed on rabbit retina in vitro, it was possible to measure O2 consumption and lactate production with the retinas under resting conditions and in different states of physiological activity. Resting energy consumption was large, as has been previously reported, and there were increases of up to 2.3 times with activity. Under some circumstances, the demands appeared to exceed the energy available. It was calculated that less than 5% of the energy generated by the retina was used for "vegetative metabolism" (i.e., for the anabolic reactions essential for viability), so that even in the resting state, the great majority of the energy usage appeared to be for function-related processes. This conclusion received further support from the finding that 50% of the energy generated was used for Na+ transport. The data obtained on retina are compared with published data on brain, which also suggest that a large fraction of the energy generated is used for function-related processes. It seems reasonable to conclude that by reversibly blocking the energy-demanding processes responsible for neurophysiological functioning, it would be possible to eliminate most of the energy requirements of CNS tissue and to reduce markedly its vulnerability to hypoperfusion.

Animals↗
PubMed1992

Energy metabolism of rabbit retina as related to function: high cost of Na+ transport.

Experiments designed to examine the energy requirements of neurophysiological function were performed on isolated rabbit retina. Function was altered by photic stimulation or by function-specific drugs, and the response of energy metabolism was assessed by simultaneous measurements of O2 consumption and lactate production. In other experiments, the supply of O2 or glucose was reduced and the effect on energy metabolism and electrophysiological function was observed. Energy requirements under control conditions in darkness were high, with O2 consumption (per gm dry wt) at 11.3 mumol min-1, with lactate production at 14.8 mumol min-1, and with the derived value for glucose consumption at 9.3 mumol min-1 and for high-energy phosphate (approximately P) generation at 82.6 mumol min-1. Energy reserves were small. Removing glucose abolished the b-wave of the electroretinogram (ERG) with a t1/2 of 1 min, but did not immediately affect O2 consumption or the PIII of the ERG. Removing O2 caused increases of up to 2.7-fold in glycolysis (Pasteur effect) and caused both PIII and b-wave to fail, with a t1/2 of about 5 min. Neurotransmission through the inner retina was supported almost entirely by glycolysis, as evidenced by large increases in lactate production in response to flashing light and decreases in response to transmitter blockers (2.3-fold overall change), with no change in O2 consumption. Phototransduction, on the other hand, was normally supported by oxidative metabolism. The dark current accounted for 41% of the retina's O2 consumption. With O2 reduced, the dark current was partially supported by glycolysis, which accounts (at least in part) for the large Pasteur effect. Na+ transport by NaK ATPase accounted for about half of all energy used, as evidenced by the response to strophanthidin, that is, for 49% of the oxidative energy and 58% of the glycolytic energy. The t1/2 for the turnover of intracellular Na+ was calculated from these data to be less than 1 min. Changes in temperature caused changes in the amplitude of light-evoked electrical responses of 6.5% per degree and caused changes in both O2 consumption and glycolysis of 6.8% per degree (Q10 = 1.9). A surprisingly large fraction of oxidative energy, corresponding to about 40% of the total energy generated, could not be assigned to phototransduction, to neurotransmission, to Na+ transport for other purposes, or to vegetative metabolism. We cannot account for its usage, but it may be related to the (previously reported) rapid turnover of the gamma-phosphate of retinal GTP, the function of which also remains unknown.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1992

Energy requirements of glutamatergic pathways in rabbit retina.

In vitro rabbit retina was used as an example of CNS tissue in experiments designed to measure the energy requirements associated with the activation of different types of glutamate receptors. Retinas were exposed to glutamate and to four analogs: kainate, 2-amino-4-phosphonobutyric acid (APB), 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX), and 2-amino-5-phosphonovaleric acid (APV). The changes in O2 consumption and lactate production were determined using a recently developed experimental system that permitted simultaneous measurements of the rates at which O2 was removed from the medium and acid was added. The glutamatergic agents had relatively little effect on oxidative metabolism, but they caused large changes in glycolysis. Kainate increased retinal lactate production by 50%, whereas APB, CNQX, and APV reduced it by 23%, 19%, and 35%, respectively. Glutamate increased lactate production by 16% when administered after APB, but decreased it by 12% when administered after CNQX. The changes in energy metabolism coincided with changes in electrophysiological function. Since the energy metabolism of many retinal cells was presumably not much affected by the glutamatergic agents, the changes measured as a percent of total retinal glycolysis must have reflected considerably larger fractional changes in the cells most affected. From the response to inhibitors, it seems probable that even under resting conditions in darkness, activity in glutamatergic pathways is responsible for more than 50% of the glycolytically derived energy used by the cells involved. It also seems probable (particularly from the response to kainate) that under some circumstances the cells' energy metabolism and/or transport capability cannot meet the requirements imposed by glutamate-induced increases in function.(ABSTRACT TRUNCATED AT 250 WORDS)

2-Amino-5-phosphonovalerate↗
PubMed1988

Metabolic flux of cyclic GMP and phototransduction in rabbit retina.

1. Rabbit retinas were isolated and subjected in vitro to shifts between light and darkness in the presence or absence of four concentrations of the phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine (IBMX). Changes in the rate of cyclic GMP hydrolysis (determined by 18O labelling of guanine nucleotide alpha-phosphoryls) and in total cyclic GMP content (determined by radioimmunoassay) were compared with the changes in the electrical potential across the retina. The experiments were designed so that the changes in potential would reflect changes in the light-sensitive conductance of the photoreceptors. 2. IBMX at 27-730 microM caused dose-related reductions in cyclic GMP hydrolysis in both light and darkness. The reductions in hydrolysis were associated with almost equal reductions in synthesis, so that there was little increase in the total content of cyclic GMP despite large changes in its metabolic flux. 3. Shifting from light (2.3 x 10(3) photons microns-2 s-1) to darkness also caused large reductions in the metabolic flux of cyclic GMP, with little increase in its total content. 4. Reductions in cyclic GMP flux were always associated with increases in the vitreous-positive transretinal potential, which was used as a measure of photoreceptor outer segment conductance, and the inverse correlation between flux and potential was closely maintained (r = 0.98) under all conditions examined. The correlation between total cyclic GMP content and transretinal potential was much less close. 5. Since IBMX and darkness acted similarly and additively, the combination of IBMX and darkness caused large decreases, of up to 21-fold, in cyclic GMP flux and large increases, of up to 23-fold, in the transretinal potential. 6. Kinetic analysis of the data indicated that the great majority (about 95%) of the light-sensitive conductance was closed under physiological conditions in darkness. 7. The data appear to be consistent with a system in which much of the cyclic GMP is bound, in which the binding is increased by light, and in which the free cyclic GMP acts co-operatively with a Hill coefficient of 3 to open outer segment conductance and to inhibit guanylate cyclase.

1-Methyl-3-isobutylxanthine↗
PubMed1988

Reduction of cellular energy requirements. Screening for agents that may protect against CNS ischemia.

Protection of the brain and spinal cord against ischemia is a goal of vast clinical importance. One approach to this objective is to reduce the tissue's functional activity in order to preserve energy for the metabolic processes that are essential to viability. Experiments to explore ways of reducing function-related energy demands were performed on isolated rabbit retina, a well-characterized model of organized adult mammalian central nervous system (CNS) tissue. The retina was maintained in a nearly physiological state in a miniature "heart-lung" apparatus. Energy metabolism (oxygen consumption and glycolysis) and electrophysiological function (determined by electroretinogram) of the in vitro retina were monitored, and their responses to a series of agents that may reduce energy requirements were determined. Large reversible reductions in O2 consumption, glycolysis, and electrophysiological function were seen in response to mild hypothermia (-3 degrees to -6 degrees C), phenytoin (Dilantin, 100 to 200 mg/kg), chlordiazepoxide (Librium, 200 microM), lithium (1 to 4 mM), Mg++ (6 to 20 mM), strophanthidin (0.15 to 0.25 microM), CO2 (25% to 30%), 2-amino-5-phosphonovaleric acid (APV, 500 microM), amiloride (1 mM), and dantrolene (1 mM). One retina was exposed simultaneously to a combination of six of these agents, which reduced its oxidative and glycolytic metabolism to less than 50% of the control level. The retina recovered metabolic and electrophysiological function after a 2 1/2-hour exposure period. Other agents tested (diphenhydramine, midazolam, nifedipine, nimodipine, and quercetin) had effects on energy metabolism and electrophysiological function that were poorly reversible. Surprisingly little effect was seen in response to general anesthetic agents (thiopental and Althesin) and other CNS depressants (chlorpromazine, ethanol, lidocaine, paraldehyde, valproic acid, and baclofen). The presumed mechanisms through which these agents reduce cellular energy requirements, as well as their potential roles in the treatment of CNS ischemia, are discussed.

Animals↗
PubMed1986

Light-induced increases in cGMP metabolic flux correspond with electrical responses of photoreceptors.

The metabolism of photoreceptor cGMP and the relationship of its light-sensitive regulation to rhodopsin photoisomerization and to the photoreceptor electrical response was examined in isolated, intact rabbit retinas. The dynamics of cGMP metabolism were assessed by measuring the rate of 18O incorporation from 18O-water into the alpha-phosphoryls of the guanine nucleotides. The photoreceptor electrical response was determined by measuring the aspartate-isolated mass receptor potential. Basal cGMP flux in dark-adapted retinas was 33 pmol cGMP X mg protein-1 X s-1 which translates into a metabolic rate in the rod outer segment (ROS) of 1.7 mM/min in ATP equivalents. Photic stimulation increased this flux as much as 4.5-fold. With continuous illumination, increasing intensity caused increments in cGMP metabolic flux to a maximum of 4.5-fold, with corresponding increases in the electrical response over the same 3-log unit intensity range. Tight coupling between activation of guanylate cyclase and phosphodiesterase was indicated by either no changes in cGMP steady state concentrations or relatively small fluctuations represented by increases of 50% at lower light intensities and a 12% decrease at one of the highest intensities. A stoichiometry of about 10,000 molecules of cGMP generated and hydrolyzed per photon absorbed was calculated for the lowest light intensity when the increment in cGMP metabolic flux per photon was maximal. Flashing light caused an increase in flux in proportion to frequency up to 1 Hz and a nearly proportional increase in the voltage time integral of the electrical response up to 0.5 Hz. This indicates that the temporal resolution, or "on"/"off" rate, of the cGMP metabolic response was as fast or faster than the temporal resolution of the electrical response. The concentration of cGMP remained relatively stable in spite of the marked acceleration of cGMP flux that occurred over the 32-fold range of frequencies tested. Taken together these results show that the light-accelerated rate of cGMP synthesis tightly coupled to hydrolysis becomes a primary energy-utilizing system in the photoreceptor and represents a response that fulfills certain of the fundamental criteria required of a metabolic event playing an essential role in phototransduction.

Animals↗
PubMed1985

Bylaws. One hospital's experience.

Dr. Porter-O'Grady, in the preceding article, discussed some implications of and a model for bylaws. The author of this article relates her institution's experience in developing nursing staff bylaws. She shares the process as well as the advantages and disadvantages of implementing a system for professional accountability and autonomy.

Constitution and Bylaws↗
PubMed1983

Earliest irreversible changes during ischemia.

The ability to synthesize new protein was used as a marker of irreversible neuronal injury in experiments with isolated rabbit retinas exposed to various types of ischemic insult. The retinal neurons were able to fully recover their protein synthetic capacity after 20 min of complete ischemic anoxia, but not after 30 min. There was better toleration to either isolated substrate deprivation or complete anoxia than to both together. Increasing extracellular Mg2+ prolonged toleration to complete ischemic-anoxia. Removing Ca2+ completely from the extracellular fluid exacerbated injury. Moreover, increasing extracellular volume improved toleration to the combined insult. This experiment suggests that injured neurons may elaborate cytotoxic compounds into the extracellular fluid. This suggestion was confirmed by further experiments demonstrating exacerbation of injury following minimum insults when the retina was incubated with other already extensively damaged tissue.

Animals↗
PubMed1981

In vitro retina as an experimental model of the central nervous system.

Methods are described for isolating adult rabbit retinal and maintaining it in a medium designed to resemble CSF. Morphologic, metabolic, nd electrophysiologic measurements obtained on the in vitro retinas showed that they remained in a nearly physiological state for at least 8 h, and even after 2 days in vitro they still exhibited a high level of metabolic activity and electrical responsiveness to light. Physiological activity was modified by photic stimulation, and data are presented to document changes in metabolism in response to the changes in function. The isolated retina appears to offer a number of unusual advantages for studying relationships between function and metabolism in organized mammalian central nervous tissue.

Amino Acids↗
PubMed1980

Protein turnover in retina.

Rabbit retinas were exposed in vitro to 0.5-h pulses of [3H]leucine or [14C]leucine. Some retinas were harvested promptly after labeling to measure synthesis. These were combined, in double-labeling experiments, with retinas that had been returned to unlabeled medium for a subsequent 1 h or 3.75 h to measure degradation. All of the proteins were solubilized, and separated according to size by gel electrophoresis. The gels were cut into 95 slices, and each slice was differentially counted. The amount of protein in the slice was estimated from the Coomassie blue staining, and its molecular weight from the distribution of molecular weight (MW) standards. Turnover rates of the various sizes of proteins were calculated from these data using certain well-defined assumptions. Retinal protein contained about 32 X 10(3) nmol of polypeptide per g, with a median MW of 27,000. Total synthesis was at the rate of 103 nmol/g of protein/h, with the most rapid synthesis in the 33,000--43,000 MW range, at 2 nmol/g/h for every 1000 increment in MW. Protein renewal averaged 0.52%/h, but varied directly (p < 0.0001) with MW, so that proteins of 10,000 MW were being renewed at about 0.1%/h and proteins of 140,000 MW at about 1.4%/h. Taken together, the measurements of fractional renewal and the measurements of degradation of the newly synthesized proteins demonstrated that each slice contained proteins with markedly different breakdown coefficients, and provided enough information to characterize the proteins in the slice in terms of a fast and slow subgroup. This analysis indicated that: breakdown coefficients varied much more than rates of synthesis and were therefore the prime determinant of the amount of each protein that was present; as MW increased, breakdown coefficients of the long-lived proteins increased (p < 0.0001), accounting in major part for the correlation between size and turnover; most staining bands were due to proteins with peculiarly long lifespans; the proteins with the slowest turnover of all appeared to be histones; there was an unusually rapid synthesis of a 138,000 MW polypeptide with a moderately short half-life (about 3 h).

Animals↗