Search PubMed⌕ Search

Biomedical subjects

M D Stern

Publications and source records attributed to M D Stern.

At least 37 records · Page 2Linked to original sources
PubMed1999

Local control models of cardiac excitation-contraction coupling. A possible role for allosteric interactions between ryanodine receptors.

In cardiac muscle, release of activator calcium from the sarcoplasmic reticulum occurs by calcium- induced calcium release through ryanodine receptors (RyRs), which are clustered in a dense, regular, two-dimensional lattice array at the diad junction. We simulated numerically the stochastic dynamics of RyRs and L-type sarcolemmal calcium channels interacting via calcium nano-domains in the junctional cleft. Four putative RyR gating schemes based on single-channel measurements in lipid bilayers all failed to give stable excitation-contraction coupling, due either to insufficiently strong inactivation to terminate locally regenerative calcium-induced calcium release or insufficient cooperativity to discriminate against RyR activation by background calcium. If the ryanodine receptor was represented, instead, by a phenomenological four-state gating scheme, with channel opening resulting from simultaneous binding of two Ca2+ ions, and either calcium-dependent or activation-linked inactivation, the simulations gave a good semiquantitative accounting for the macroscopic features of excitation-contraction coupling. It was possible to restore stability to a model based on a bilayer-derived gating scheme, by introducing allosteric interactions between nearest-neighbor RyRs so as to stabilize the inactivated state and produce cooperativity among calcium binding sites on different RyRs. Such allosteric coupling between RyRs may be a function of the foot process and lattice array, explaining their conservation during evolution.

Algorithms↗
PubMed1999

Calcium release flux underlying Ca2+ sparks of frog skeletal muscle.

An algorithm for the calculation of Ca2+ release flux underlying Ca2+ sparks (Blatter, L.A., J. Hüser, and E. Ríos. 1997. Proc. Natl. Acad. Sci. USA. 94:4176-4181) was modified and applied to sparks obtained by confocal microscopy in single frog skeletal muscle fibers, which were voltage clamped in a two-Vaseline gap chamber or permeabilized and immersed in fluo-3-containing internal solution. The performance of the algorithm was characterized on sparks obtained by simulation of fluorescence due to release of Ca2+ from a spherical source, in a homogeneous three-dimensional space that contained components representing cytoplasmic molecules and Ca2+ removal processes. Total release current, as well as source diameter and noise level, was varied in the simulations. Derived release flux or current, calculated by volume integration of the derived flux density, estimated quite closely the current used in the simulation, while full width at half magnitude of the derived release flux was a good monitor of source size only at diameters >0. 7 micrometers. On an average of 157 sparks of amplitude >2 U resting fluorescence, located automatically in a representative voltage clamp experiment, the algorithm reported a release current of 16.9 pA, coming from a source of 0.5 micrometer, with an open time of 6.3 ms. Fewer sparks were obtained in permeabilized fibers, so that the algorithm had to be applied to individual sparks or averages of few events, which degraded its performance in comparable tests. The average current reported for 19 large sparks obtained in permeabilized fibers was 14.4 pA. A minimum estimate, derived from the rate of change of dye-bound Ca2+ concentration, was 8 pA. Such a current would require simultaneous opening of between 8 and 60 release channels with unitary Ca2+ currents of the level recorded in bilayer experiments. Real sparks differ from simulated ones mainly in having greater width. Correspondingly, the algorithm reported greater spatial extent of the source for real sparks. This may again indicate a multichannel origin of sparks, or could reflect limitations in spatial resolution.

Algorithms↗
PubMed1999

Effects of different levels of methionine and ruminally undegradable protein on the amino acid profile of effluent from continuous culture fermenters.

Eight dual-flow continuous culture fermenters were used to study the effects of feeding different levels of methionine and ruminally undegradable protein (RUP) on the amino acid (AA) profile of protein residue after microbial fermentation. Four isocaloric (1.76 Mcal/kg of NE(l)) and isonitrogenous (17.7% CP) diets were formulated based on NRC recommendations for a 630-kg cow producing 42 kg/d of milk. Two dietary treatments, diets HmLr (high methionine, low RUP) and HmHr (high methionine, high RUP), were formulated to meet, but not exceed, more than 140% of each estimated individual AA requirement for milk production. Diets LmLr (low methionine, low RUP) and LmHr (low methionine, high RUP) were formulated to meet the estimated individual AA requirement except for methionine. Diets HmLr and LmLr contained 35% RUP, and diets HmHr and LmHr contained 43% RUP. Digestion of DM, OM, and NDF was not affected (P>.05) by treatments. Tyrosine, cystine, and threonine ranked as the most resistant AA to microbial fermentation, whereas methionine and lysine ranked among the most degradable. In general, microbial degradation of individual AA followed the same pattern as CP degradation, being higher in diets HmLr and LmLr and lower in diets HmHr and LmHr. Because true microbial digestion of methionine, cystine, tyrosine, and isoleucine was reduced in the presence of high levels of methionine, we concluded that supplementation of dietary methionine could increase flows of these amino acids to the duodenum.

Amino Acids↗
PubMed1999

Effects of type of carbohydrate supplementation to lush pasture on microbial fermentation in continuous culture.

Eight single-flow continuous culture fermenters were used to study the effects of the type of energy source on ruminal N utilization from high quality pasture. The four dietary treatments included high quality grass and legume pasture alone (50:50; wt/wt), pasture plus soybean hulls, pasture plus beet pulp, and pasture plus corn. Diets supplemented with additional sources of energy (soybean hulls, beet pulp, and corn) were isocaloric but differed in the type and rate of carbohydrate fermentation. Energy supplements constituted 45% of the total dietary dry matter and were fed twice daily at 12-h intervals in place of pasture, which is characteristic of grain feeding at milking when animals are in a grazing situation. Energy supplementation reduced pH, NH3 N flow, and NH3 N concentration and increased bacterial N flow (as a percentage of N intake). The supplementation of corn and soybean hulls resulted in the highest microbial N flow (as a percentage of N intake). Corn had a tendency to reduce fiber digestion because of excessively low NH3 N concentrations. Beet pulp was similar to corn in that it decreased NH3 N concentrations. Supplementation of soybean hulls resulted in a more synchronized fermentation, greater volatile fatty acid production, and greater fiber digestion. Nitrogen utilization by microbes was maximized by supplementation with soybean hulls or corn twice a day. With diets based on pasture, it may be more important to improve bacterial N flow and bacterial utilization of N than to maximize the efficiency of bacterial protein synthesis because better utilization of N by ruminal microorganisms results in higher bacterial N flow and higher fiber digestion.

Ammonia↗
PubMed1998

Termination of Ca2+ release by a local inactivation of ryanodine receptors in cardiac myocytes.

In heart, a robust regulatory mechanism is required to counteract the regenerative Ca2+-induced Ca2+ release from the sarcoplasmic reticulum. Several mechanisms, including inactivation, adaptation, and stochastic closing of ryanodine receptors (RyRs) have been proposed, but no conclusive evidence has yet been provided. We probed the termination process of Ca2+ release by using a technique of imaging local Ca2+ release, or "Ca2+ spikes", at subcellular sites; and we tracked the kinetics of Ca2+ release triggered by L-type Ca2+ channels. At 0 mV, Ca2+ release occurred and terminated within 40 ms after the onset of clamp pulses (0 mV). Increasing the open-duration and promoting the reopenings of Ca2+ channels with the Ca2+ channel agonist, FPL64176, did not prolong or trigger secondary Ca2+ spikes, even though two-thirds of the sarcoplasmic reticulum Ca2+ remained available for release. Latency of Ca2+ spikes coincided with the first openings but not with the reopenings of L-type Ca2+ channels. After an initial maximal release, even a multi-fold increase in unitary Ca2+ current induced by a hyperpolarization to -120 mV failed to trigger additional release, indicating absolute refractoriness of RyRs. When the release was submaximal (e.g., at +30 mV), tail currents did activate additional Ca2+ spikes; confocal images revealed that they originated from RyRs unfired during depolarization. These results indicate that Ca2+ release is terminated primarily by a highly localized, use-dependent inactivation of RyRs but not by the stochastic closing or adaptation of RyRs in intact ventricular myocytes.

Animals↗
PubMed1998

Direct measurement of SR release flux by tracking 'Ca2+ spikes' in rat cardiac myocytes.

1. Ca2+ release flux across the sarcoplasmic reticulum (SR) during cardiac excitation-contraction coupling was investigated using a novel fluorescence method. Under whole-cell voltage-clamp conditions, rat ventricular myocytes were dialysed with a high concentration of EGTA (4.0 mM, 150 nM free Ca2+), to minimize the residence time of released Ca2+ in the cytoplasm, and a low-affinity, fast Ca2+ indicator, Oregon Green 488 BAPTA-5N (OG-5N; 1.0 mM, Kd approximately 31 microM), to optimize the detection of localized high [Ca2+] in release site microdomains. Confocal microscopy was employed to resolve intracellular [Ca2+] at high spatial and temporal resolution. 2. Analytical and numerical analyses indicated that, under conditions of high EGTA concentration, the free [Ca2+] change is the sum of two terms: one major term proportional to the SR release flux/Ca2+ influx, and the other reflecting the running integral of the released Ca2+. 3. Indeed, the OG-5N transients in EGTA-containing cells consisted of a prominent spike followed by a small pedestal. The OG-5N spike closely resembled the first derivative (d[Ca2+]/dt) of the conventional Ca2+ transient (with no EGTA), and mimicked the model-derived SR Ca2+ release function reported previously. In SR Ca2+-depleted cells, the OG-5N transient also closely followed the waveform of L-type Ca2+ current (ICa). Using ICa as a known source of Ca2+ influx, SR flux can be calibrated in vivo by a linear extrapolation of the ICa-elicited OG-5N signal. 4. The OG-5N image signal was localized to discrete release sites at the Z-line level of sarcomeres, indicating that the local OG-5N spike arises from 'Ca2+ spikes' at transverse (T) tubule-SR junctions (due to the imbalance between calcium ions entering the cytosol and the buffer molecules). 5. Both peak SR release flux and total amount of released Ca2+ exhibited a bell-shaped voltage dependence. The temporal pattern of SR release also varied with membrane voltage: Ca2+ release was most synchronized and produced maximal peak release flux (4.2 mM s-1) at 0 mV; in contrast, maximal total release occurred at -20 mV (71 versus 61 microM at 0 mV), but the localized release signals were partially asynchronous. Since the maximal conventional [Ca2+] transient and contraction were elicited at 0 mV, it appears that not only the amount of Ca2+ released, but also the synchronization among release sites affects the whole-cell Ca2+ transient and the Ca2+-myofilament interaction.

Algorithms↗
PubMed1998

A simple numerical model of calcium spark formation and detection in cardiac myocytes.

The elementary events of excitation-contraction coupling in heart muscle are Ca2+ sparks, which arise from one or more ryanodine receptors in the sarcoplasmic reticulum (SR). Here a simple numerical model is constructed to explore Ca2+ spark formation, detection, and interpretation in cardiac myocytes. This model includes Ca2+ release, cytosolic diffusion, resequestration by SR Ca2+-ATPases, and the association and dissociation of Ca2+ with endogenous Ca2+-binding sites and a diffusible indicator dye (fluo-3). Simulations in a homogeneous, isotropic cytosol reproduce the brightness and the time course of a typical cardiac Ca2+ spark, but underestimate its spatial size (approximately 1.1 micron vs. approximately 2.0 micron). Back-calculating [Ca2+]i by assuming equilibrium with indicator fails to provide a good estimate of the free Ca2+ concentration even when using blur-free fluorescence data. A parameter sensitivity study reveals that the mobility, kinetics, and concentration of the indicator are essential determinants of the shape of Ca2+ sparks, whereas the stationary buffers and pumps are less influential. Using a geometrically more complex version of the model, we show that the asymmetric shape of Ca2+ sparks is better explained by anisotropic diffusion of Ca2+ ions and indicator dye rather than by subsarcomeric inhomogeneities of the Ca2+ buffer and transport system. In addition, we examine the contribution of off-center confocal sampling to the variance of spark statistics.

Aniline Compounds↗
PubMed1998

Mitochondrial calcium transporting pathways during hypoxia and reoxygenation in single rat cardiomyocytes.

OBJECTIVE: Mitochondrial [Ca2+] ([Ca2+]m) rises in parallel with cytosolic [Ca2+] ([Ca2+]c) following ATP-depletion rigor contracture induced by hypoxia in isolated cardiomyocytes. We investigated the pathways involved in the hypoxia induced changes in [Ca2+]m by using known inhibitors of mitochondrial Ca2+ transport, namely ruthenium red, an inhibitor of the Ca2+ uniporter (the normal influx route) and clonazepam, an inhibitor of Na+/Ca2+ exchange, (the normal efflux route). METHODS: [Ca2+]m was determined from indo-1/am loaded rat myocytes where the cytosolic fluorescence signal had been quenched by superfusion with Mn2+. [Ca2+]c was measured by loading myocytes with indo-1 pentapotassium salt during the isolation procedure. Cells were placed in a specially developed chamber for induction of hypoxia and reoxygenated 40 min after rigor development. RESULTS: 50% of control cells hypercontracted upon reoxygenation; this correlated with a [Ca2+]m or [Ca2+]c higher than approximately 350 nM at the end of rigor. Clonazepam completely abolished the rigor-induced rise in [Ca2+]m but not [Ca2+]c. On reoxygenation [Ca2+]m increased over the first 5 min and remained elevated whereas [Ca2+]c fell. In the presence of ruthenium red a dramatic increase in [Ca2+]m occurred 5-10 min after rigor development (the indo-1 fluorescence signal was saturated); [Ca2+]c also increased but to a lesser extent. On reoxygenation, [Ca2+]m fell rapidly even though cells hypercontracted and [Ca2+]c remained elevated. CONCLUSIONS: During hypoxia following rigor development Ca2+ uptake into mitochondria occurs largely via the Na+/Ca2+ exchanger rather than the Ca2+ uniporter whereas on reoxygenation the transporters resume their normal directionality.

Animals↗
PubMed1998

Evaluation of selected mathematical approaches to the kinetics of protein degradation in situ.

A linear model, two mathematical nonlinear models, and a curve-peeling procedure were used to estimate rate and extent of ruminal CP degradation of meat and bone meal (MBM) and soybean meal (SBM) from data obtained using the in situ Dacron polyester bag technique. Most of the values for extent of CP degradation of MBM were lowest when determined using curve peeling or the nonlinear models. In general, rates and extents of CP degradation of MBM estimated using the linear model and including ruminal incubations up to 12 h were greater than those obtained with the linear model and including ruminal incubations up to 24 h or up to 72 h. In addition, the models ranked the MBM samples differently for rate and extent of CP degradation. The results of the lack-of-fit test indicated that the linear model was inappropriate for estimating rate of degradation of MBM. However, CP degradation for SBM could be described by the linear model if long ruminal incubation times (greater than 48 h) were included in the calculations. Regression analyses were conducted to evaluate various compositional characteristics as predictors of CP degradation for MBM. Most of the correlation coefficients between CP degradation and the same independent variables were greater when the nonlinear models and curve peeling were used compared with the linear model. In general, the correlation coefficients between extent of CP degradation and the independent variables obtained with the linear model increased as the duration of ruminal incubations included in the model increased. Lysine concentrations, followed by CP solubility and ash content, were the best predictors of ruminal degradation of MBM protein. When using a specific mathematical model to predict CP degradation, analysis of residuals vs fitted and lack-of-fit tests should be performed to assess the validity of the model to describe the degradation patterns of the protein source under consideration. Also, long (at least 48 h) ruminal incubation times may be needed to correctly describe the pattern of CP degradation for MBM.

Animal Feed↗
PubMed1997

Partial depletion of sarcoplasmic reticulum calcium does not prevent calcium sparks in rat ventricular myocytes.

1. The exact nature of calcium sparks in the heart remains highly controversial. We sought to determine whether calcium sparks arise from a single or multiple calcium release channels/ ryanodine receptors in the sarcoplasmic reticulum (SR). If their genesis involves a calcium-coupled recruitment of multiple channels, calcium sparks might be abolished by a modest depletion of SR calcium (because of the decrease in unitary calcium flux and hence a decrease in the gain of local calcium-induced calcium release). If, on the other extreme, calcium sparks are produced despite severe SR depletion, the single-channel origin will be preferred. 2. Spontaneous calcium sparks were studied in rat ventricular myocytes using confocal microscopy and the fluorescent calcium probe fluo-3. A computer algorithm was developed to count and measure objectively calcium sparks in linescan images. 3. Thapsigargin (25-150 nM) depleted caffeine-releasable SR calcium by up to 64%, in a dose- and time-dependent manner, without altering the resting cytosolic calcium level. During SR depletion, calcium sparks were robustly observed, albeit at reduced frequency (> or = 30% of control) and amplitude (> or = 60% of control). 4. Due to the reduced detectability of small sparks against noise background, the observed data would overestimate reduction in spark frequency but underestimate amplitude reduction. After correction for this detection bias, we found that the spark frequency was independent of SR load, whereas the amplitude was proportional to load. 5. We conclude that, although spark amplitude depends on SR filling status, the frequency of spark generation is independent of SR calcium load, and therefore independent of the local calcium release rate. This implies that sparks are single-channel events, or collective events that are well above threshold for local regeneration. Additionally, our results suggest that intraluminal SR calcium, at normal or low loads, does not play a major role in the regulation of on-gating of the ryanodine receptor.

Adamantane↗
PubMed1997

Inhibition of mitochondrial calcium efflux by clonazepam in intact single rat cardiomyocytes and effects on NADH production.

The aims of this study were to determine: (i) whether clonazepam and CGP37157, which inhibit the Na+/Ca2+ exchanger of isolated mitochondria, could inhibit mitochondrial Ca2+ efflux in intact cells; and (ii) whether any sustained increase in mitochondrial [Ca2+] ([Ca2+]m) could alter mitochondrial NADH levels. [Ca2+]m was measured in Indo-1/AM loaded rat ventricular myocytes where the cytosolic fluorescence signal was quenched by superfusion with Mn2+. NADH levels were determined from cell autofluorescence. Upon exposure of myocytes to 50 nM norepinephrine (NE) and a stimulation rate of 3 Hz, [Ca2+]m increased from 59 +/- 3 nM to a peak of 517 +/- 115 nM (n = 8) which recovered rapidly upon return to low stimulation rate (0.2 Hz) and washout of NE. In the presence of clonazepam, the peak increase in [Ca2+]m was 937 +/- 192 nM (n = 5) which remained elevated at 652 +/- 131 nM upon removal of the stimulus. CGP37157 in some cells did give the same inhibition of mitochondrial Ca2+ efflux as clonazepam, but the effect was inconsistent since not all cells were capable of following the stimulation rate in presence of this compound. NADH levels increased upon exposure to rapid stimulation in the presence of NE alone and recovered upon return to low stimulation rates, whereas in clonazepam treated cells the recovery of NADH was prevented. We conclude that clonazepam is an effective inhibitor of mitochondrial [Ca2+] efflux in intact cells and also maintains the increase in NADH levels which occurs upon rapid stimulation of cells.

Animals↗
PubMed1997

Local control model of excitation-contraction coupling in skeletal muscle.

This is a quantitative model of control of Ca release from the sarcoplasmic reticulum in skeletal muscle, based on dual control of release channels (ryanodine receptors), primarily by voltage, secondarily by Ca (Ríos, E., and G. Pizarro. 1988. 3:223-227). Channels are positioned in a double row array of between 10 and 60 channels, where exactly half face voltage sensors (dihydropyridine receptors) in the transverse (t) tubule membrane (Block, B.A., T. Imagawa, K.P. Campbell, and C. Franzini-Armstrong. 1988. 107:2587-2600). We calculate the flux of Ca release upon different patterns of pulsed t-tubule depolarization by explicit stochastic simulation of the states of all channels in the array. Channels are initially opened by voltage sensors, according to an allosteric prescription (Ríos, E., M. Karhanek, J. Ma, A. González. 1993. 102:449-482). Ca permeating the open channels, diffusing in the junctional gap space, and interacting with fixed and mobile buffers produces defined and changing distributions of Ca concentration. These concentrations interact with activating and inactivating channel sites to determine the propagation of activation and inactivation within the array. The model satisfactorily simulates several whole-cell observations, including kinetics and voltage dependence of release flux, the "paradox of control," whereby Ca-activated release remains under voltage control, and, most surprisingly, the "quantal" aspects of activation and inactivation (Pizarro, G., N. Shirokova, A. Tsugorka, and E. Ríos. 1997. 501:289-303). Additionally, the model produces discrete events of activation that resemble Ca sparks (Cheng, H., M.B. Cannell, and W.J. Lederer. 1993. 262:740-744). All these properties result from the intersection of stochastic channel properties, control by local Ca, and, most importantly, the one dimensional geometry of the array and its mesoscopic scale. Our calculations support the concept that the release channels associated with one face of one junctional t-tubule segment, with its voltage sensor, constitute a functional unit, termed the "couplon." This unit is fundamental: the whole cell behavior can be synthesized as that of a set of couplons, rather than a set of independent channels.

Animals↗
PubMed1997

Calcium in close quarters: microdomain feedback in excitation-contraction coupling and other cell biological phenomena.

Researchers have made good progress in unraveling the molecular mechanisms of excitation-contraction (EC) coupling in striated muscle. Despite this progress, paradoxes abound. In skeletal muscle, the existence of a mechanical coupling between membrane charge movement and activation of sarcoplasmic reticulum (SR) release channels is essentially established, but the contribution of Ca(2+)-induced Ca2+ release (CICR) to the transient and steady-state components of Ca2+ release remains controversial. In cardiac muscle, the role of CICR as the primary mechanism of EC coupling is well established, but the stability and tight coupling between membrane Ca2+ current and release are paradoxical. Answers may lie in microdomain issues, and the examination of discrete elementary release events, although quantitative treatments are needed. This review explores the theoretical and experimental methods used and the observations made in the study of microdomain Ca2+.

Animals↗
PubMed1997

Measurement of mitochondrial calcium in single living cardiomyocytes by selective removal of cytosolic indo 1.

The aim of the present study was to determine whether, in indo 1 acetoxymethyl ester (AM)-loaded rat cardiomyocytes, it was possible to remove cytosolic but not mitochondrial indo 1 by promoting loss of cytosolic indo 1 through plasma membrane anion pumps (which are blocked by probenecid). Isolated rat ventricular myocytes were loaded with indo 1-AM under conditions (15 min at 30 degrees C) in which about half of the dye is located within mitochondria. Cells were then maintained at 25 degrees C for 2.5 h followed by incubation at 37 degrees C for 1.5 h. After this "heat treatment," the myocyte fluorescence signal was 44% of the value of cells measured before heat treatment, and loss of fluorescence was prevented by 1 mM probenecid. The remaining fluorescence was shown to originate from mitochondria, since 1) Ca2+ uptake and efflux could be inhibited by ruthenium red and clonazepam, respectively, and 2) low concentrations of digitonin, which release only cytosolic marker enzymes, decreased fluorescence of untreated myocytes but had little effect on the fluorescence signal of heat-treated cells. We conclude that heat treatment selectively removes cytosolic indo 1, leaving a signal due to mitochondrial indo 1 only.

Animals↗
PubMed1997

Myocyte adaptation to chronic hypoxia and development of tolerance to subsequent acute severe hypoxia.

Studies in animal models and humans suggest that myocardium may adapt to chronic or intermittent prolonged episodes of reduced coronary perfusion. Stable maintenance of partial flow reduction is difficult to achieve in experimental models; thus, in vitro cellular models may be useful for establishing the mechanisms of adaptation. Since moderate hypoxia is likely to be an important component of the low-flow state, isolated adult rat cardiac myocytes were exposed to 1% O2 for 48 hours to study chronic hypoxic adaptation. Hypoxic culture did not reduce cell viability relative to normoxic controls but did enhance glucose utilization and lactate production, which is consistent with an anaerobic pattern of metabolism. Lactate production remained transiently increased after restoration of normal O2 tension. Myocyte contractility was reduced (video-edge analysis), as was the amplitude of the intracellular Ca2+ transient (indo 1 fluorescence) in hypoxic cells. Relaxation was slowed and was accompanied by a slowed decay of the Ca2+ transient. These changes were not due to alterations in the action potential. Tolerance to subsequent acute severe hypoxia occurred in cells cultured in 1% O2 and was manifested as a delay in the time to full ATP-depletion rigor contracture during severe hypoxia and enhanced morphological recovery of myocytes at reoxygenation. The latter was still seen after normalization of the data for the prolonged time to rigor, suggesting a multifactorial basis for tolerance. An intervening period of normoxic exposure before subsequent acute severe hypoxia did not result in loss of tolerance but rather increased the delay to subsequent ATP depletion rigor. Cellular glycogen was preserved during chronic hypoxic exposure and increased after the restoration of normal O2 tension. As mitochondrial cytochromes should be fully oxygenated at levels well below 1% O2, hypoxic adaptation may be mediated by a low-affinity O2-sensing process. Thus, adaptations that occur during prolonged periods of moderate hypoxia are proposed to poise the myocyte in a better position to tolerate impending episodes of severe O2 deprivation.

Action Potentials↗
PubMed1997

Alternative techniques for measuring nutrient digestion in ruminants.

Because in vivo measurement of nutrient digestion in the rumen and small intestine requires ruminally and intestinally cannulated animals that are expensive, labor-intensive, and subject to error associated with markers and inherent animal variation, alternative techniques have been developed. Researchers have proposed various in situ or in vitro procedures for estimating ruminal and small intestinal nutrient digestion. This review summarizes these alternative techniques.

Animals↗
PubMed1996

"Adaptive" behavior of ligand-gated ion channels: constraints by thermodynamics.

The calcium-induced calcium release channel of the cardiac sarcoplasmic reticulum has been reported to inactivate in a novel manner (termed "adaptation"), which permits reactivation by exposure to successively higher concentrations of calcium. I examined the limitations placed by thermodynamics on the possible kinetic mechanisms for such behavior. The mechanism suggested by Gyorke and Fill, in which the affinity of a calcium-binding site decreases during adaptation, is not thermodynamically feasible for a passive system, but requires an external input of free energy. Possible sources of such energy are 1) metabolic energy, which is excluded by the fact that adaptation was observed in isolated channels in the absence of ATP, or 2) coupling of ion permeation to gating, for which there is currently no evidence. I derived a general limit on the thermodynamic feasibility of a sequence of channel activations and adaptations, irrespective of channel kinetics, from the requirement that the free energy must decrease during the spontaneous evolution of the system from the state existing immediately after a step increase in [Ca2+] to the state of maximum open probability that follows. The opening of the channel must involve an increase in free energy, which must be compensated by the free energy released by the incremental binding of calcium. This requirement leads to a complicated system of inequalities, which was simplified and manipulated algebraically into the form of a linear programming problem. Numerical solution of this problem showed that the sequence of adaptations of the SR channel observed by Gyorke and Fill requires the presence of at least 10 calcium-binding sites on the channel if it is to occur in the absence of exogenous sources of free energy. This indicates either that a large number of calcium-binding sites participate in the regulation of the SR calcium release channel, or that the existing data are significantly flawed with respect to the low open probability in the resting state, the importance of "calcium spike" artifacts from flash photolysis, or both.

Adaptation, Physiological↗