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Biomedical subjects

E Ríos

Publications and source records attributed to E Ríos.

At least 19 recordsLinked to original sources
PubMed1999

Amplitude distribution of calcium sparks in confocal images: theory and studies with an automatic detection method.

Determination of the calcium spark amplitude distribution is of critical importance for understanding the nature of elementary calcium release events in striated muscle. In the present study we show, on general theoretical grounds, that calcium sparks, as observed in confocal line scan images, should have a nonmodal, monotonic decreasing amplitude distribution, regardless of whether the underlying events are stereotyped. To test this prediction we developed, implemented, and verified an automated computer algorithm for objective detection and measurement of calcium sparks in raw image data. When the sensitivity and reliability of the algorithm were set appropriately, we observed highly left-skewed or monotonic decreasing amplitude distributions in skeletal muscle cells and cardiomyocytes, confirming the theoretical predictions. The previously reported modal or Gaussian distributions of sparks detected by eye must therefore be the result of subjective detection bias against small amplitude events. In addition, we discuss possible situations when a modal distribution might be observed.

Algorithms↗
PubMed1999

Unitary Ca2+ current through cardiac ryanodine receptor channels under quasi-physiological ionic conditions.

Single canine cardiac ryanodine receptor channels were incorporated into planar lipid bilayers. Single-channel currents were sampled at 1-5 kHz and filtered at 0.2-1.0 kHz. Channel incorporations were obtained in symmetrical solutions (20 mM HEPES-Tris, pH 7.4, and pCa 5). Unitary Ca2+ currents were monitored when 2-30 mM Ca2+ was added to the lumenal side of the channel. The relationship between the amplitude of unitary Ca2+ current (at 0 mV holding potential) and lumenal [Ca2+] was hyperbolic and saturated at approximately 4 pA. This relationship was then defined in the presence of different symmetrical CsCH3SO3 concentrations (5, 50, and 150 mM). Under these conditions, unitary current amplitude was 1.2 +/- 0.1, 0.65 +/- 0.1, and 0.35 +/- 0.1 pA in 2 mM lumenal Ca2+; and 3.3 +/- 0.4, 2.4 +/- 0. 2, and 1.63 +/- 0.2 pA in 10 mM lumenal Ca2+ (n > 6). Unitary Ca2+ current was also defined in the presence of symmetrical [Mg2+] (1 mM) and low [Cs+] (5 mM). Under these conditions, unitary Ca2+ current in 2 and 10 mM lumenal Ca2+ was 0.66 +/- 0.1 and 1.52 +/- 0.06 pA, respectively. In the presence of higher symmetrical [Cs+] (50 mM), Mg2+ (1 mM), and lumenal [Ca2+] (10 mM), unitary Ca2+ current exhibited an amplitude of 0.9 +/- 0.2 pA (n = 3). This result indicates that the actions of Cs+ and Mg2+ on unitary Ca2+ current were additive. These data demonstrate that physiological levels of monovalent cation and Mg2+ effectively compete with Ca2+ as charge carrier in cardiac ryanodine receptor channels. If lumenal free Ca2+ is 2 mM, then our results indicate that unitary Ca2+ current under physiological conditions should be <0.6 pA.

Algorithms↗
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↗
PubMed1998

Local calcium release in mammalian skeletal muscle.

1. Fluo-3 fluorescence associated with Ca2+ release was recorded with confocal microscopy in single muscle fibres mechanically dissected from fast twitch muscle of rats or frogs, voltage clamped in a two Vaseline-gap chamber. 2. Interventions that elicited Ca2+ sparks in frog skeletal muscle (low voltage depolarizations, application of caffeine) generated in rat fibres images consistent with substantial release from triadic regions, but devoid of resolvable discrete events. Ca2+ sparks were never observed in adult rat fibres. In contrast, sparks of standard morphology were abundant in myotubes from embryonic mice. 3. Depolarization-induced gradients of fluorescence between triadic and surrounding regions (which are proportional to Ca2+ release flux) peaked at about 20 ms and then decayed to a steady level. Gradients were greater in frog fibres than in rat fibres. The ratio of peak over steady gradient (R) was steeply voltage dependent in frogs, reaching a maximum of 4.8 at -50 mV (n = 7). In rats, R had an essentially voltage-independent value of 2.3 (n = 5). 4. Ca2+-induced Ca2+ release, resulting in concerted opening of several release channels, is thought to underlie Ca2+ sparks and the peak phase of release in frog skeletal muscle. A diffuse 'small event' release, similar to that observed in these rats, is also present in frogs and believed to be directly activated by voltage. The present results suggest that in these rat fibres there is little contribution by CICR to Ca2+ release triggered by depolarization, and a lack of concerted channel opening.

Aniline Compounds↗
PubMed1998

Molecular cloning and functional expression of a skeletal muscle dihydropyridine receptor from Rana catesbeiana.

In skeletal muscle the dihydropyridine receptor is the voltage sensor for excitation-contraction coupling and an L-type Ca2+ channel. We cloned a dihydropyridine receptor (named Fgalpha1S) from frog skeletal muscle, where excitation-contraction coupling has been studied most extensively. Fgalpha1S contains 5600 base pairs coding for 1688 amino acids. It is highly homologous with, and of the same length as, the C-truncated form predominant in rabbit muscle. The primary sequence has every feature needed to be an L-type Ca2+ channel and a skeletal-type voltage sensor. Currents expressed in tsA201 cells had rapid activation (5-10 ms half-time) and Ca2+-dependent inactivation. Although functional expression of the full Fgalpha1S was difficult, the chimera consisting of Fgalpha1S domain I in the rabbit cardiac Ca channel had high expression and a rapidly activating current. The slow native activation is therefore not determined solely by the alpha1 subunit sequence. Its Ca2+-dependent inactivation strengthens the notion that in rabbit skeletal muscle this capability is inhibited by a C-terminal stretch (Adams, B., and Tanabe, T. (1997) J. Gen. Physiol. 110, 379-389). This molecule constitutes a new tool for studies of excitation-contraction coupling, gating, modulation, and gene expression.

Amino Acid Sequence↗
PubMed1998

Inactivation of gating currents of L-type calcium channels. Specific role of the alpha 2 delta subunit.

In studies of gating currents of rabbit cardiac Ca channels expressed as alpha 1C/beta 2a or alpha 1C/beta 2a/alpha 2 delta subunit combinations in tsA201 cells, we found that long-lasting depolarization shifted the distribution of mobile charge to very negative potentials. The phenomenon has been termed charge interconversion in native skeletal muscle (Brum, G., and E. Ríos. 1987. J. Physiol. (Camb.). 387:489-517) and cardiac Ca channels (Shirokov, R., R. Levis, N. Shirokova, and E. Ríos. 1992. J. Gen. Physiol. 99:863-895). Charge 1 (voltage of half-maximal transfer, V1/2 approximately 0 mV) gates noninactivated channels, while charge 2 (V1/2 approximately -90 mV) is generated in inactivated channels. In alpha 1C/beta 2a cells, the available charge 1 decreased upon inactivating depolarization with a time constant tau approximately 8, while the available charge 2 decreased upon recovery from inactivation (at -200 mV) with tau approximately 0.3 s. These processes therefore are much slower than charge movement, which takes <50 ms. This separation between the time scale of measurable charge movement and that of changes in their availability, which was even wider in the presence of alpha 2 delta, implies that charges 1 and 2 originate from separate channel modes. Because clear modal separation characterizes slow (C-type) inactivation of Na and K channels, this observation establishes the nature of voltage-dependent inactivation of L-type Ca channels as slow or C-type. The presence of the alpha 2 delta subunit did not change the V1/2 of charge 2, but sped up the reduction of charge 1 upon inactivation at 40 mV (to tau approximately 2 s), while slowing the reduction of charge 2 upon recovery (tau approximately 2 s). The observations were well simulated with a model that describes activation as continuous electrodiffusion (Levitt, D. 1989. Biophys. J. 55:489-498) and inactivation as discrete modal change. The effects of alpha 2 delta are reproduced assuming that the subunit lowers the free energy of the inactivated mode.

Algorithms↗
PubMed1997

Small event Ca2+ release: a probable precursor of Ca2+ sparks in frog skeletal muscle.

1. Fluo-3 fluorescence associated with Ca2+ release was recorded with confocal microscopy in single muscle fibres. Clamp depolarization to -65 or -60 mV elicited Ca2+ sparks with amplitudes and spatial widths distributed approximately normally, with mean values of 0.79 of resting fluorescence and 0.8 micron (S.D., 0.17 and 0.2 micron; n = 193), respectively. Given these distributions, events of amplitude less than 0.45 or width less than 0.4 micron are unlikely to be sparks. 2. Low voltage depolarization (-72 mV) elicited only one spark per triad every 6 s, but generated a relative increase in fluorescence at triads of 0.05. This increase must therefore have been due to events smaller than sparks. 3. The variance/mean ratio of triadic fluorescence gradients averaged 0.11 at low voltages and increased severalfold at the higher voltages at which sparks appeared, indicating the existence of at least two event amplitudes. 4. Tetracaine (200 microM) reversibly abolished sparks and the early peak of Ca2+ release at all voltages. In its presence, discrete events were smaller than the spark criterion, and triadic gradients had a variance/mean ratio of 0.11. 5. The phenylalkylamine D600 (2 microM) reduced release at all voltages, abolishing sparks and the peak of Ca2+ release at low but not at high voltages. 6. The parallel abolition of all sparks and the peak of Ca2+ release indicates that both phenomena are activated by Ca2+. The restoration of sparks by voltage in D600 suggests that release in small events provides the trigger Ca2+ for activation of sparks.

Anesthetics, Local↗
PubMed1997

'Quantal' calcium release operated by membrane voltage in frog skeletal muscle.

1. Ca2+ transients and Ca2+ release flux were determined optically in cut skeletal muscle fibres under voltage clamp. 'Decay' of release during a depolarizing pulse was defined as the difference between the peak value of release and the much lower steady level reached after about 100 ms of depolarization. Using a double-pulse protocol, the inactivating effect of release was measured by 'suppression', the difference between the peak values of release in the test pulse, in the absence and presence of a conditioning pulse that closely preceded the test pulse. 2. The relationship between decay and suppression was found to follow two simple arithmetic rules. Whenever the conditioning depolarization was less than or equal to the test depolarization, decay in the conditioning release was approximately equal to suppression of the test release. Whenever the conditioning depolarization was greater than that of the test, suppression was complete, i.e. test release was reduced to a function that increased monotonically to a steady level. The steady level was the same with or without conditioning. 3. These arithmetic rules suggest that inactivation of Ca2+ release channels is strictly and fatally linked to their activation. More than a strict linkage, however, is required to explain the arithmetic properties. 4. The arithmetic rules of inactivation result in three other properties that are inexplicable with classical models of channel gating: constant suppression, incremental inactivation and increment detection. These properties were first demonstrated for inositol trisphosphate (IP3)-sensitive channels and used to define IP3-induced release as quantal. In this sense, it can now be stated that skeletal muscle Ca2+ release is activated by membrane voltage in a quantal manner. 5. For both classes of intracellular Ca2+ channels, one explanation of the observations is the existence of subsets of channels with different sensitivities (to voltage or agonist dose). In an alternative explanation, channels are identical, but have a complex repertoire of voltage- or dose-dependent responses.

Animals↗
PubMed1997

Sarcoplasmic reticulum Ca2+ release flux underlying Ca2+ sparks in cardiac muscle.

Discrete events of Ca2+ release from the sarcoplasmic reticulum (SR) have been described in cardiac, skeletal, and smooth muscle. In skeletal muscle these release events originate at individual channels. In cardiac muscle, however, it remains a question of debate whether localized Ca2+ release transients, termed Ca2+ sparks, originate from single release channels or multiple channels clustered in close vicinity. Generalizing methods used earlier to describe cell-averaged Ca2+ release, we derived, as a function of space and time, the flux of Ca2+ release that underlies Ca2+ sparks. Using the method to analyze spontaneous sparks recorded with confocal microscopy in dissociated cat atrial cells, we obtained in most cases single sparks of Ca2+ release that appear to originate from approximately 1-microm-wide regions. In many cases, doublets, triplets, and greater groups of release sparks were observed. This multiplicity, the estimated release flux magnitude, and existing data on the structure of junctions between SR and plasmalemma suggest that individual release sparks result from the opening of multiple Ca2+ release channels clustered within discrete SR junctional regions.

Animals↗
PubMed1997

Ion-dependent inactivation of barium current through L-type calcium channels.

It is widely believed that Ba2+ currents carried through L-type Ca2+ channels inactivate by a voltage-dependent mechanism similar to that described for other voltage-dependent channels. Studying ionic and gating currents of rabbit cardiac Ca2+ channels expressed in different subunit combinations in tsA201 cells, we found a phase of Ba2+ current decay with characteristics of ion-dependent inactivation. Upon a long duration (20 s) depolarizing pulse, IBa decayed as the sum of two exponentials. The slow phase (tau approximately 6 s, 21 degrees C) was parallel to a reduction of gating charge mobile at positive voltages, which was determined in the same cells. The fast phase of current decay (tau approximately 600 ms), involving about 50% of total decay, was not accompanied by decrease of gating currents. Its amplitude depended on voltage with a characteristic U-shape, reflecting reduction of inactivation at positive voltages. When Na+ was used as the charge carrier, decay of ionic current followed a single exponential, of rate similar to that of the slow decay of Ba2+ current. The reduction of Ba2+ current during a depolarizing pulse was not due to changes in the concentration gradients driving ion movement, because Ba2+ entry during the pulse did not change the reversal potential for Ba2+. A simple model of Ca(2+) -dependent inactivation (Shirokov, R., R. Levis, N. Shirokova, and F., Ríos. 1993. J. Gen. Physiol. 102:1005-1030) robustly accounts for fast Ba2+ current decay assuming the affinity of the inactivation site on the alpha 1 subunit to be 100 times lower for Ba2+ than Ca2+.

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

[Experience with radical prostatectomy as treatment of localized cancer of the prostate at the Fundación Santa Fe de Bogotá, Colombia].

OBJECTIVE: To analyze the experience of the Fundación Santa Fe de Bogotá with radical prostatectomy in the treatment of localized prostatic cancer. METHODS: A retrospective study was conducted on 108 patients with localized carcinoma of the prostate stage T1-T2NxM0 submitted to radical prostatectomy from 1989 to 1994. RESULTS: Preoperatively, 50% of the patients had a PSA < 10 ng/ml and 14% had values that fell within the normal ranges of 0-4 mg/ml. A family history of prostate cancer was detected in 9.3% of the patients. The prostate cancer was clinically understaged in 75% of the patients, particularly those with stages T2a and T2b, and less significantly in those with stage T2c. Considering only those patients in whom the pathological staging had disclosed the cancer was not localized, this incidence accounted for 52% (n = 57). The presence of surgical margins was approximately 36%. The tumor recurrence rate was 26.9% and the complication rate was 6.8%. CONCLUSION: The relatively low complication rate in the present series shows that radical prostatectomy is a safe procedure that achieves good results if the cases are carefully selected and the diagnostic test are widely utilized.

Age Distribution↗
PubMed1996

Activation of Ca2+ release by caffeine and voltage in frog skeletal muscle.

1. Using a fast flow, computer-controlled, two-Vaseline-gap chamber, single muscle fibres were subjected to 'pulses' of caffeine at Ca2+ releasing concentrations, combined with voltage-clamp depolarizations, while monitoring intracellular [Ca2+]. 2. Ca2+ release flux elicited by caffeine reached 2.5 mM s-1, or less, after 3 s of exposure, then decayed to zero. The caffeine-releasable pool of sarcoplasmic reticulum (SR) Ca2+ was 2.9 +/- 0.4 mM (mean +/- S.E.M., n = 10). 3. In parallel with release induced by caffeine, release induced by voltage pulses applied during a caffeine exposure increased in the first second of exposure, then decreased, to abolition after 5 s. 4. The amount of Ca2+ releasable by depolarizing pulses was always equal to the amount of Ca2+ in the caffeine-releasable pool. Therefore, there is a single releasable Ca2+ pool. This pool is well stirred-it takes much more time to lose its Ca2+ by release than to diffusionally homogenize its [Ca2+]. Its depletion explains quantitatively the decay of release induced by caffeine or voltage during an exposure to caffeine. 5. A 1.5 s pulse to 10 mV, applied during exposure to caffeine, resulted in large Ca2+ release and, upon repolarization, termination of the caffeine-induced release. This is similar to repolarization-induced stop of caffeine contracture (RISC) in embryonic murine myoballs. The permeability elicited by caffeine (ratio of flux to calcium in the releasable pool) was not affected by depolarizing pulses. Therefore, the mechanism of the RISC-like effect was Ca2+ depletion. 6. Caffeine-induced release did not depend on the holding potential. 7. Whether caffeine was present or not, release activated by voltage remained always under voltage control, ending rapidly upon repolarization. A depolarizing pulse induced a release permeability with an early peak, followed by decay to a steady level. Caffeine (10 mM) shifted the mid-activation voltage of both peak and steady components by -15 mV and increased the steepness of their voltage dependence by 15%. The maximum permeability increased by 30% for the peak and 25% for the steady component (n = 5). These results neither support nor disprove the hypothesis that the peak of Ca2+ release is activated by Ca2+. 8. The similar potentiation by caffeine of both components of release, the continued ability of voltage to control release in the presence of caffeine, and its failure to alter caffeine-induced permeability indicate that caffeine and the voltage sensor enhance independently the channel's tendency to open.

Animals↗
PubMed1996

Caffeine enhances intramembranous charge movement in frog skeletal muscle by increasing cytoplasmic Ca2+ concentration.

1. Currents of intramembranous charge movement were recorded, together with intracellular [Ca2+], in single muscle fibres subjected to voltage-clamp depolarization and 'pulses' of extracellular solution with a Ca2+ release-inducing concentration of caffeine (10 mM). 2. When caffeine was present prior to and during the voltage pulses, the charge transferred by pulses to between -60 and -40 mV increased by about 40%. 3. In fibres depleted of Ca2+ in the sarcoplasmic reticulum (SR), caffeine had no effect on charge transfer or kinetics. 4. Whenever the prior exposure to caffeine resulted in a large elevation in [Ca2+]i at the start of the depolarizing pulse, there was an increase in I beta, the monotonically decaying component of charge movement. When the presence of caffeine enhanced Ca2+ release induced by the pulse, there was increase in I gamma, the hump-like component. 5. The charge transferred during a pulse to -50 mV increased with time of exposure to caffeine. Ca2+ release induced by the voltage pulse grew during the first second of caffeine exposure, then decreased with longer exposure time. The enhancement of charge transfer by caffeine was therefore not due to the increase in Ca2+ release caused by the drug. 6. The increase in charge transfer was a uniform, monotonically increasing function of the [Ca2+]i attained at the end of the voltage pulse. 7. Charge transfer, as a function of [Ca2+]i, pulse voltage and time, was simulated with a model, used previously, in which Ca2+ binds to intracellular sites and increases the electrical potential near the voltage sensors. Two sites were needed to fit the observations, with dissociation constants of 60 nM and 2 to 10 microM. 8. In the presence of caffeine, the voltage-driven movement of a given amount of intra-membranous charge resulted in greater activation of release permeability. 9. All effects of caffeine observed in this and the preceding paper could be explained assuming a single action: caffeine increases the tendency of the release channels to open. This results in opening of closed channels and an increase in their susceptibility to activation by the voltage sensors.

Animals↗
PubMed1996

Ca2+ release from the sarcoplasmic reticulum compared in amphibian and mammalian skeletal muscle.

Puzzled by recent reports of differences in specific ligand binding to muscle Ca2+ channels, we quantitatively compared the flux of Ca2+ release from the sarcoplasmic reticulum (SR) in skeletal muscle fibers of an amphibian (frog) and a mammal (rat), voltage clamped in a double Vaseline gap chamber. The determinations of release flux were carried out by the "removal" method and by measuring the rate of Ca2+ binding to dyes in large excess over other Ca2+ buffers. To have a more meaningful comparison, the effects of stretching the fibers, of rapid changes in temperature, and of changes in the Ca2+ content of the SR were studied in both species. In both frogs and rats, the release flux had an early peak followed by fast relaxation to a lower sustained release. The peak and steady values of release flux, Rp and Rs, were influenced little by stretching. Rp in frogs was 31 mM/s (SEM = 4, n = 24) and in rats 7 +/- 2 mM/s (n = 12). Rs was 9 +/- 1 and 3 +/- 0.7 mM/s in frogs and rats, respectively. Transverse (T) tubule area, estimated from capacitance measurements and normalized to fiber volume, was greater in rats (0.61 +/- 0.04 microns-1) than in frogs (0.48 +/- 0.04 micron-1), as expected from the greater density of T tubuli. Total Ca in the SR was estimated as 3.4 +/- 0.6 and 1.9 +/- 0.3 mmol/liter myoplasmic water in frogs and rats. With the above figures, the steady release flux per unit area of T tubule was found to be fourfold greater in the frog, and the steady permeability of the junctional SR was about threefold greater. The ratio Rp/Rs was approximately 2 in rats at all voltages, whereas it was greater and steeply voltage dependent in frogs, going through a maximum of 6 at -40 mV, then decaying to approximately 3.5 at high voltage. Both Rp and Rs depended strongly on the temperature, but their ratio, and its voltage dependence, did not. Assuming that the peak of Ca2+ release is contributed by release channels not in contact with voltage sensors, or not under their direct control, the greater ratio in frogs may correspond to the relative excess of Ca2+ release channels over voltage sensors apparent in binding measurements. From the marked differences in voltage dependence of the ratio, as well as consideration of Ca(2+)-induced release models, we derive indications of fundamental differences in control mechanisms between mammalian and amphibian muscle.

Animals↗
PubMed1996

Leukaemia of natural killer cell large granular lymphocyte type with HLA-DR-CD16-CD56bright+ phenotype.

The case is reported of a 45 year old woman with the rare leukaemia of natural killer cell large granular lymphocyte (NK/ LGL) type. Cytometric analysis of leukaemic blasts showed that they were positive for CD2, CD38, and CD56 antigens but negative for a series of antigens including CD3, CD7, CD16, and HLA-DR. Rearrangements of the beta T cell receptor, and heavy and kappa immunoglobulin genes were not detected and neither were chromosomal abnormalities. Leukaemic blasts developed NK cytotoxicity. The patient failed to respond to aggressive chemotherapy and died three months after diagnosis. The lack of expression of HLA-DR is an extraordinary characteristic of this case, as all cases of acute NK cell leukaemias described to date expressed HLA-DR. The immunophenotype observed in the NK cell leukaemic blasts may represent the counterpart of a hypothetical normal cell precursor in an early stage of ontogenic NK cell development.

Acute Disease↗