Biomedical subjects

M Mazzanti

Publications and source records attributed to M Mazzanti.

At least 37 records · Page 2Linked to original sources

PubMed1996

Cytoskeletal control of rectification and expression of four substates in cardiac inward rectifier K+ channels.

Cardiac inward rectifiers may have a three-barrel channel structure, based on evidence for three substates in single-channel recordings. However, some reports indicate four substates, a feature more compatible with the four-subunit structure for which there is evidence in cloned voltage-activated K+ channels. Here we show that although the fourth is easily missed, inward rectifier channels have four substates whose expression is controlled by intracellular Ca(2+) ions. Fourth substate openings also appear after rectification loss in intracellular divalent caution-free solution. We find that this process is accelerated by cytochalasin, a microfilament disrupter. Cytochalasin also abolishes Ca(2+), but not Mg(2+),-induced rectification by restoring fourth substate openings. Thus, cytoskeletal elements control Ca(2+)-dependent substate expression and rectification in native inwardly rectifying K+ channels.

Animals↗

PubMed1995

Automatic quantification of ejection fraction from gated myocardial perfusion SPECT.

UNLABELLED: We have developed a completely automatic algorithm to quantitatively measure left ventricular ejection fraction (LVEF) from gated 99mTc-sestamibi myocardial perfusion SPECT images. METHODS: The algorithm operates in the three-dimensional space and uses gated short-axis image volumes. It segments the left ventricle (LV), estimates and displays endocardial and epicardial surfaces for all gating intervals in the cardiac cycle, calculates the relative left ventricular cavity volumes and derives the global EF from the end-diastolic and end-systolic volume, all without operator interaction. The algorithm for measuring LVEF was tested in 65 clinical patients undergoing 16-interval and 8-interval rest-gated SPECT and validated against first-pass radionuclide ventriculography. RESULTS: Automatic segmentation and contouring of the LV was successful in 65/65 (100%) of the studies. Agreement between EFs measured from 8-interval gated SPECT and EFs calculated from first-pass data was high (y = 2.44 + 1.03x, r = 0.909, p < 0.001, s.e.e. = 6.87). Agreement between EF values measured from 16-interval and 8-interval gated SPECT was excellent (y = -2.7 + 0.97x, r = 0.988, p < 0.001, s.e.e. = 2.65), the latter being on average lower by 3.71 percentage points. CONCLUSION: Our automatic method is rapid and highly agrees with conventional radionuclide measurements of EF, thus providing clinically useful additional information to complement myocardial perfusion studies.

Algorithms↗

PubMed1995

Automatic reorientation of three-dimensional, transaxial myocardial perfusion SPECT images.

UNLABELLED: We developed a completely automatic technique to reorient transaxial images into short-axis (oblique) myocardial perfusion SPECT images. METHODS: The algorithm starts by isolating (segmenting) the left ventricle (LV) myocardium using a combination of iterative clusterification and rule-based location/size/shape criteria. The three-dimensional, mid-myocardial LV surface is initially estimated as the locus of the trilinearly interpolated maxima for the count profiles originating from the center of mass of the segmented LV. The final mid-myocardial surface is obtained by iteratively applying this process, incorporating additional constraints of shape and texture and using the nonsegmented, nonthresholded transaxial image to obtain information on hypoperfused areas of the myocardium. It is then fitted to an ellipsoid, of which the major axis is assumed to represent the long axis of the LV, and the three-dimensional image volume is resliced perpendicularly to it. RESULTS: The algorithm was retrospectively applied to 400 dual-isotope studies (200 rest 201TI, 200 stress 99mTc-sestamibi) from 200 consecutive patients. Segmentation was successful in 394/400 (98.5%) of the patients. The reproducibility of computer-based reorientation was perfect and significantly better than either intraobserver or interobserver reproducibility. CONCLUSION: Automatic reorientation offers the potential for consistently faster and more accurate image processing and analysis and is an important step towards totally operator-less management of myocardial perfusion SPECT data.

Algorithms↗

PubMed1994

Inactivation of single Ca2+ channels in rat sensory neurons by extracellular Ca2+.

1. Single Ca2+ channels conducting 20 mM Ba2+ from adult rat dorsal root ganglion cells were characterized using the two-electrode patch-clamp technique configuration. 2. Channels demonstrating specific characteristics of conductance, voltage dependence and dihydropyridine sensitivity were classified as high-threshold or L-type Ca2+ channels. 3. Mean single-channel current in 20 mM Ba2+ did not show inactivation, but inactivation occurred when using Ca2+ as a permeating ion. 4. Stimulus protocols were delivered alternately in the cell-attached and whole-cell electrode, while recording single-channel activity and total Ca2+ current simultaneously. 5. A mean single-channel Ba2+ current from a stimulated patch did not show inactivation. However, stimulation of a physiological whole-cell Ca2+ current induced a marked inactivation of mean single-channel Ba2+ current. 6. Complete Ca2+ current block by the addition of 200 microM Cd2+ in the external solution removed single-channel inactivation in patches stimulated through a whole-cell electrode.

Animals↗

PubMed1994

ATP-dependent ionic permeability on nuclear envelope in in situ nuclei of Xenopus oocytes.

The nuclear envelope represents a structural and functional barrier between cytoplasm and nucleoplasm. Small molecules and solutes passively cross the nuclear envelope, whereas the transport of large proteins and RNA requires metabolic energy. Using in situ Xenopus oocyte nuclei, we characterized ATP-dependent ionic permeabilities on the external surface of the envelope. The presence, but not necessarily the hydrolysis, of ATP is crucial to maintaining the channels in an open state. Localization of the ionic channels is still unclear. From morphologic and current kinetics data, we suggest a relation between the ionic channels and the nuclear pores. We try, in this way, to explain the apparent contradiction between the presence of ion-selective channels in parallel with large aqueous pores on the nuclear envelope. Under this hypothesis, variations in the metabolic energy content of the cytoplasm would induce nucleocytoplasmic passive exchanges. The distribution and movement of charged particles across the nuclear envelope may influence many cytoplasmic functions. Regulation of the current by ATP could play an important role in hormonal stimulation, divalent ion permeation into the nucleus, and cell cycle mechanisms.

Adenosine Triphosphate↗

PubMed1993

Identification of a nucleo-cytoplasmic ionic pathway by osmotic shock in isolated mouse liver nuclei.

The observation that the nuclear envelope outer membrane contains ion channels raises the question of whether these conductances communicate between the cytosol and the nuclear envelope cisternae or between the cytosol and the cytoplasm. Failure to detect large, nonselective holes using the patch-clamp technique has led to the speculation that ion channels and nuclear pores are in fact the same. In this paper we present evidence that the ionic channel, recorded in isolated liver nuclei with the patch-clamp configuration of "nucleus-attached," spans the double membrane of the envelope, providing a direct contact between nucleoplasm and cytoplasm.

Animals↗

PubMed1992

Na channels that remain open throughout the cardiac action potential plateau.

In this paper we report the direct measurement of rare Na channel events that occur during the cardiac action potential, viz., channels that open at the upstroke and remain open throughout the plateau and early repolarization phase. The technique we use allows us to record channel activity and action potentials at the same time; thus, we are certain of when the Na channels open and when they finally close. The slow Na channels have the same voltage dependence, single-channel conductance, and TTX sensitivity as the fast Na channels, and they conduct Li. It therefore seems likely that the fast and the slow currents flow through the same channel. If this interpretation is correct, then the Na channel not only initiates the action potential but also helps to maintain its plateau. It is possible that the slow Na currents represent a separate collection of channels rather than a low-probability state of the fast Na channels. Regardless of which interpretation is correct, the present experiments allow us to assess the effect of the slow currents on action potential shape and on sustained Na entry.

Action Potentials↗

PubMed1992

Effect of firing rate on the calcium permeability in adult neurons during spontaneous action potentials.

Calcium channels in neurons mediate a wide variety of essential functions. In addition to contributing to action potential shape, they furnish a substrate that acts as an intracellular second messenger. This study shows that the shape of the neuronal action potential has characteristics that promote long openings of L-type (high threshold) calcium channels. We also present evidence that a change in the firing rate of isolated neurons modulates gating of single calcium channels. This mechanism could be important in modulating neuron excitability and providing a rise in intracellular Ca, when needed.

Action Potentials↗

PubMed1992

[Effects of aerobic training in patients with moderate chronic heart failure].

UNLABELLED: To evaluate the effects of a program of moderate intensity in patients (pts) with clinically stable chronic heart failure (CHF), we studied 20 pts (18M, 2F, mean age 61 years) with dilated cardiomyopathy, Weber Class B, ejection fraction (EF) < 40% and aerobic capacity of 16 +/- 2 ml/kg/min on cardiopulmonary exercise testing. We randomly assigned pts to 2 groups, a training group (T, 10 pts) and a control group (C, 10 pts), similar for anatomical and clinical characteristics, group T underwent a thrice weekly, 8 week-long ambulatory program of aerobic activity, beginning at 40% of maximal oxygen uptake. At the end, in Group T we observed a significant increase of exercise tolerance (+45%; p < 0.005), peak oxygen uptake (VO2 max) (+20%; p < 0.001), anaerobic ventilatory threshold (AT) (+37%; p < 0.005), lactate threshold (+36%; p < 0.005), peak heart rate (< 10%; p < 0.01) and of peak systolic pressure (+12%; p < 0.007); and a significant reduction in resting heart rate (-17%; p < 0.005), resting diastolic pressure (-11%; p < 0.005), plasma lactate (LA) at rest (-26%; p < 0.01), at peak (-21%; p < 0.005) and at recovery (-22%; p < 0.005), plasma norepinephrine (NE) both at rest (-38%; p < 0.005) and at peak (-13%; p < 0.005) and of plasma epinephrine (E) (-38%; p < 0.005; -32%, p < 0.001, respectively). We observed no change in EF at the end in both groups nor any untoward cardiac effects during training. We didn't note any correlation between AT and venous oxygen saturation (r = 0.15; p = 0.65) changes at the end. The increase in peak VO2 after training was not correlated to any AT increase (r = 0.12; p = 0.72). We observed, however, a significant correlation between lactate threshold and AT changes after training (r = 0.81; p = 0.005) and between LA and resting, submaximal and peak NE (r = 0.89; p = 0.005) and E (r = 0.78; p = 0.007) changes at the end of training. CONCLUSIONS: a) in pts with clinically stable CHF a program of aerobic activity well tolerated in terms of frequency, intensity and duration may determine a significant increase in exercise tolerance, aerobic capacity, AT and LA threshold, and a significant decrease in plasma LA, NE and E at rest, submaximal and peak levels; b) in our opinion, such modifications are partly determined by a delay in lactate accumulation and partly by sympathetic tone lowering, and permit us to underline the concept that physical inactivity may provoke important peripheral changes that, in turn, may reduce exercise tolerance and aerobic capacity, by creating a vicious circle difficult to break by medical therapy alone.

Anaerobic Threshold↗

PubMed1991

Ion channels in murine nuclei during early development and in fully differentiated adult cells.

The nuclear envelope functions as a selective barrier between nucleus and cytoplasm. During cycles of cell division the nuclear envelope repeatedly disassembles and re-associates. Presumably, each cycle re-establishes the functional and structural integrity of the nuclear envelope. After repeated rounds of cell division, as occurs during differentiation, the selectivity and configuration of the envelope may change. We compare the ionic conductance and the nuclear pore density in four types of murine nuclei: germinal vesicles in oocytes, pronuclei in zygotes, nuclei from two-cell blastomeres, and somatic cell nuclei from the liver. A large-conductance ion channel is present in all nuclear envelopes. Liver cell nuclei have a greater number of these channels than those from earlier developmental stages, and they also have a higher density of nuclear pores. In this article we hypothesize an association between the ion channels and the nuclear pores.

Animals↗

PubMed1991

Chloride channels in the nuclear membrane.

Chloride-selective ion channels were measured from isolated rat liver nuclei. Single ion channel currents were recorded in both "nuclear-attached" and in excised patches in the inside-out configuration of the patch-clamp technique. Two types of chloride conductance were defined, a large conductance (150 pS; iCl,N) channel with complex kinetics and multiple substates, and a second smaller conductance (58 pS;ICln) channel sensitive to block by ATP. The channels were inhibited by pharmacological agents known to block chloride channels and were insensitive to internal and external changes in calcium and magnesium. Presumably the channels reside in the external membrane of the nuclear double membrane and may mediate charge balance in the release and uptake of calcium from the perinuclear space.

4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid↗

PubMed1991

Gating of L-type Ca2+ channels in embryonic chick ventricle cells: dependence on voltage, current and channel density.

1. L-type calcium channels in embryonic chick heart ventricle have voltage-dependent, time-variant kinetics when they conduct inward currents carried by 20 mM-Ba2+. Depolarizing the membrane from -20 to 20 mV increases mean open time from 1.4 to 4.2 ms. Mean open time increases monotonically with voltage. The single-channel conductance, 18 +/- 2 pS, is approximately linear over this voltage range, and the extrapolated reversal potential is 38 +/- 5 mV. 2. In cell-attached patches with five or more L-type Ca2+ channels in the patch, the currents elicited by 500 ms depolarizing steps, from a -80 mV holding potential, inactivate rapidly and have large tail currents. In the same patch, currents from a -40 mV holding potential are smaller, inactivate more slowly, and have practically no tail currents. 3. In cell-attached patches containing one of two L-type Ca2+ channels, currents from -80 or -40 mV are virtually identical, and they are similar to the currents from multichannel patches held at -40 mV. 4. The voltage-dependent, time-variant kinetics of individual L-type Ca2+ channels are unaltered if the patch is removed from the cell and forms an inside-out configuration. In these experiments the internal membrane was bathed with an artificial, intracellular-like solution containing no phosphorylating enzymes or substrates. 5. Cells bathed in 20 mM-Ba2+ solutions and held at -80 mV have currents with an early phase that inactivates in tens of milliseconds, a late phase that inactivates in hundreds of milliseconds, and a large, slow tail current. Currents from -40 mV have only the late phase and practically no tails. However, if the maximum current is less than 0.1 pA pF-1, records from either -80 or -40 mV are virtually identical, and they are similar to currents from cells with higher channel density held at -40 mV. Furthermore, if cells are stimulated before full recovery from inactivation, the reduced current is accompanied by slower inactivation. 6. Whole-cell currents in 1.5 mM-Ca2+ solutions are entirely abolished by addition of 20 microM-nifedipine, and they are enhanced 2-3 times by addition of 30 microM-cyclic AMP and 3 mM-ATP to the whole-cell recording electrode. The whole-cell currents in 20 mM-Ba2+ solutions are also completely blocked by 20 microM-nifedipine, regardless of kinetics or holding potential. Thus, by definition, the cells we are studying contain only L-type channels.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

PubMed1991

Clinical and instrumental evaluation of the family of a patient with arrhythmogenic right ventricle dysplasia.

UNLABELLED: The aim of the present study is a clinical and instrumental evaluation of the family of a patient with arrhythmogenic right ventricle dysplasia (ARVD), initially complicated by recurrent ventricular tachycardia and later by congestive cardiac failure. Because the probability of familial involvement is very high in those cases described in the literature, the authors evaluated the patient's 3 children: 2 daughters, twenty-four and twenty years old, respectively, and a twenty-one-year-old son; his niece, thirty-one years old; and her sons three and eight years old. The authors did not evaluate his fifty-six-year-old sister, because she was affected by rheumatic mitral valve disease. All in the study were asymptomatic, and clinical examination did not show any pathologic findings. Rest ECG was normal in all cases, and the exercise stress test (Bruce protocol) showed normal functional capacity. Holter recordings were normal without arrhythmias; chest x rays showed normal cardiothoracic ratio and cardiac morphologic volume. Two-dimensional echocardiography and pulsed and continuous wave Doppler demonstrated normal sizes of cardiac chambers and normal function and morphology of cardiac valves. Nevertheless, in 4 cases (66.5%) the right ventricles showed an apical bulging with normal systolic thickness, without hypokinetic or akinetic areas or diverticular outpouchings. The authors did not find those abnormalities in 5 control subjects who were studied in the same way. CONCLUSIONS: In 4 family members (66.5%) of a patient with ARVD progressed to cardiac congestive failure, the authors found anomalies of right ventricle morphology: apical bulging, not revealed in a control group, and an absence of symptoms.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

PubMed1990

Ion channels in the nuclear envelope.

Cell nuclei are capable of partitioning a wide variety of molecules from the cytosol, including macromolecules such as proteins and RNA, and smaller peptides, amino acids, sugars and Na+ and K+ ions, all of which can be accumulated in or excluded from the nuclear domain. There are two mechanisms behind this compartmentalization: selective retention of freely diffusible molecules, and selective entry through the nuclear envelope. It is generally accepted that the nuclear envelope restricts only the larger molecules. Here we apply the patch-clamp technique to isolated murine pronuclei and show that the nuclear envelope contains K(+)-selective channels which have multiple conductance states, the maximal conductance being 200 pS. These channels, which contribute to the nuclear membrane potential, may be important in balancing the charge carried by the movement of macromolecules in and out of the nucleus.

Animals↗

PubMed1990

Ca modulates outward current through IK1 channels.

Inward-rectifier channels in cardiac cells (IK1) stabilize the resting membrane potential near the K equilibrium potential. Here we investigate the role of IK1 in the regulation of action potentials and link this to the influx of Ca during beating. Inward Ca current alters the open-channel probability of outward IK1 current. Thus Ca ions depolarize cells not only by carrying an inward current but also by blocking an outward current.

Action Potentials↗

PubMed1990

Ca channel gating during cardiac action potentials.

How do Ca channels conduct Ca ions during the cardiac action potential? We attempt to answer this question by applying a two-microelectrode technique, previously used for Na and K currents, in which we record the patch current and the action potential at the same time (Mazzanti, M., and L. J. DeFelice. 1987. Biophys. J. 12:95-100, and 1988. Biophys. J. 54:1139-1148; Wellis, D., L. J. DeFelice, and M. Mazzanti. 1990. Biophys. J. 57:41-48). In this paper, we also compare the action currents obtained by the technique with the step-protocol currents obtained during standard voltage-clamp experiments. Individual Ca channels were measured in 10 mM Ca/1 Ba and 10 mM Ba. To describe part of our results, we use the nomenclature introduced by Hess, P., J. B. Lansman, and R. W. Tsien (1984. Nature (Lond.). 311:538-544). With Ba as the charge carrier, Ca channel kinetics convert rapidly from long to short open times as the patch voltage changes from 20 to -20 mV. This voltage-dependent conversion occurs during action potentials and in step-protocol experiments. With Ca as the charge carrier, the currents are brief at all voltages, and it is difficult to define either the number of channels in the patch or the conductance of the individual channels. Occasionally, however, Ca-conducting channels spontaneously convert to long-open-time kinetics (in Hess et al., 1984, notation, mode 2). When this happens, which is about once in every 100beats, there usually appears to be only one channel in the patch. In this rare configuration, the channel is open long enough to measure its conductance in 10 Ca/ 1 Ba. The value is 8-10 pS, which is about half the conductance in Ba. Because the long openings occur so infrequently with Ca as the charge carrier, they contribute negligibly to the average Ca current at any particular time during an action potential. However, the total number of Ca ions entering during these long openings may be significant when compared to the number entering by the more usual kinetics.

Action Potentials↗

PubMed1990

Outward sodium current in beating heart cells.

This article is a study of the fast Na current during action potentials. We have investigated the outward Na current (Mazzanti, M., and L.J. DeFelice. 1987. Biophys. J. 52:95-100) in more detail, and we have asked whether it goes through the same channels associated with the rapid depolarization phase of action potentials. We address the question by patch clamping single, spontaneously beating, embryonic chick ventricle cells, using two electrodes to record the action potential and the patch current simultaneously. The chief limitation is the capacitive current, and in this article we describe a new method to subtract it. Varying the potential and the Na concentration in the patch pipette, and fitting the corrected currents to a standard model (Ebihara, L., and E.A. Johnson. 1980. Biophys. J. 32:779-790), provides evidence that the outward current is carried by the same channels that conduct the inward current. We compare the currents in beating cells to currents in nonbeating cells using whole-cell and cell-attached patch clamp recordings. The latter tend to show more positive Na reversal potentials, with the implication that internal Na is higher in beating cells. We propose that the plateau of the action potential, which is partly due to an inward Ca current, exceeds Na action current reversal potentials, and that this driving force gives rise to an outward movement of Na ions. The existence of such a current would imply that the fast repolarization phase after the upstroke of cardiac action potentials is partly due to the Na action current.

Action Potentials↗

PubMed1990

Potassium channels and the repolarization of cardiac cells.

What is the contribution of a particular potassium current to the repolarization of cardiac myocytes? The traditional answer to this question requires clamping the cells with step voltages, finding models that describe how individual currents depend on voltage and time, driving these models with action potentials to calculate the action currents, and evaluating the contribution of each pathway to repolarization from the action currents. Another method is to measure the action currents directly from beating cells. We isolated potassium channels in cell-attached patches and averaged the current over many beats. The average channel current, mean value of i(t), is a miniature version of the action current through corresponding channels in the membrane outside the patch. The time integral of this current, scaled by channel density, N, and membrane capacity, C, is the contribution of that particular pathway to the action potential: Vi(t) = -Ni integral of to mean value of i(u) du/C Using this procedure, we have found that the delayed rectifier, IK, turns on virtually without delay following the upstroke of the action potential and gradually declines during the plateau and repolarization phases, having nearly the shape of the action potential itself. The inward rectifier, IKl, may conduct little current during the plateau and is under the control of internal Ca. The traditional method of measuring action currents from step voltage-clamp records gives qualitatively similar results. Differences may arise because factors other than voltage modulate potassium currents in beating cells.

Animals↗