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Biomedical subjects

B A Kotsias

Publications and source records attributed to B A Kotsias.

At least 19 recordsLinked to original sources
PubMed2000

Chloride channels in toad skeletal muscle fibers.

Chloride currents were measured in short lumbricalis fibers of toads (Bufo arenarum) with voltage and patch clamp techniques. For the availability of chloride currents we applied a double-pulse technique in voltage-clamped fibers. When the test pulse was preceded by a positive prepulse, the initial current was larger than with a negative prepulse and exhibited a different rate of decline to its steady-state value. At the single-channel level we found that in most of the experiments with symmetrical 110 mM NaCl solutions, two levels of conductance, 20 ("small channel") and 360 pS ("maxi channel"), occurred with the highest probabilities. The openings of the maxi channels were more frequent at potentials close to 0 mV, whereas for the small channels the openings were at negative potentials. In contrast with the results with the macroscopic currents, a change of 2 orders of magnitude in the pH, from 7.3 to 5, had only minor effects on the channels' conductance. As with some other anion channels, the selectivity of the channels described here is low, the p(Cl)/p(Na) ratio being 1.9 and 3.7 for the small and maxi Cl(-) channels, respectively. The behavior of these Cl(-) channels with a relative high Na(+) permeability could contribute to the relatively low resting membrane potential of the lumbricalis fibers measured in the standard 110 mM NaCl solution.

Animals↗
PubMed2000

[Past and present of Medicina (Buenos Aires)].

To celebrate the 60th anniversary of Medicina (Buenos Aires) an International Symposium was held at the National Academy of Medicine of Buenos Aires on the 6-7th of October 1999, under the title of Clinical investigation in the next millennium. This meeting was a success as evidenced by the 376 registered attendants. Sixty years of uninterrupted publication is an uncommon feat in our midst and this could be achieved on the basis of a number of factors which include, the initiative of those who founded the journal, the unfailing motivation and dedication of the Editorial Board and primarily the authors who have trusted us with their manuscripts. Of the many important papers published, we have selected a few which proved to be milestones in the development of Argentine biomedicine. It is to be hoped that the future will bring an increase in our impact index through more and even better papers eventually reflecting the authentic scientific value of our country.

Argentina↗
PubMed1999

Effects of DIDS, a disulfonic stilbene derivative, on chloride movements in toad skeletal muscles.

In order to investigate the characteristics of the movement of Cl- ions in toad skeletal muscles we decided to study the relative membrane permeabilities of chloride and nitrate and the effects of DIDS (4,4'-diisothyocyanatostilbene-2,2'-disulphonate) upon the hyperpolarizations produced in muscle fibers when chloride or nitrate ions rapidly replace impermeant sulphate ions in the external solution. For experiments where membrane potential changes were recorded in response to sudden changes in extracellular solutions, small bundles from the semitendinosus muscles were used. We showed that DIDS reduced in a reversible manner the Cl- permeability (pCl) in toad skeletal muscle fibers. The results supporting this conclusion were the following. First, a diminished hyperpolarization in response to a sudden exposure of the fibers to a solution containing Cl-. In these experiments DIDS reduced the pCl/pK ratio to 5.5 from a control value of 12. Second, a smaller transient of the resting potential when [Cl]o was changed from 120 to 30 mM and vice versa.

4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid↗
PubMed1997

Blockade of the inward rectifier potassium currents by zinc and nickel ions in voltage-clamped toad muscles.

The inward rectifier is one of the voltage-sensitive K+ channels present in several tissues: Its conductance increases under hyperpolarization and decreases with depolarization. In this work we studied the effects of Zn2+ and Ni2+ (5-30 mM) on the macroscopic K+ current through the inward rectifier system. The experiments were performed in the short muscle fibers of the lumbricalis muscle of toads with a two-microelectrode voltage clamp technique. The fibers were equilibrated in a control solution containing 68 mM K2SO4 and then exposed to Zn2+ or Ni2+. We found that both cations reduced in a reversible manner the current carried by K+ ions, and this reduction was prevented by decreasing the external pH of the solution (pH 5). The blockade of current was slightly dependent on the membrane potential and time independent. Two mechanisms may be involved in the blocking action of these cations: Zn2+ and Ni2+ may either be blocking the pore of the channels or acting at a regulatory binding site on the extracellular surface in an unspecified manner.

Animals↗
PubMed1997

[Ionic-channel diseases].

This review illustrates several hereditary diseases caused by mutations in genes which encode various ion channels activated by voltage or neurotransmitters. Many physiological processes depend upon the proper functioning of plasma membrane ion channels and this is most apparent in absorptive and secretory epithelia, and in electrically excitable tissues such as nerve and muscle. By combining the information from electrophysiological recordings with molecular biological techniques, further insight can be gained into the gene expression and protein structure of ionic channels. This combination has resulted in a structure-function analysis revealing the molecular substructures of the ionic channels responsible for the processes of permeation and selectivity of activation and inactivation and different types of block. Using molecular biologic tools, these abnormal channels can be identified and their molecular defects defined. Advances in these areas now provide the basis for a rational approach to the classification and treatment of these disorders of membrane excitation.

Genetic Diseases, Inborn↗
PubMed1996

Chloride current in toad skeletal muscle and its modification by the histidine-modifying reagent diethylpyrocarbonate.

Cl- currents were measured in short fibres in the toad lumbricalis muscle with a two-microelectrode voltage clamp. Membrane Cl- conductance increased markedly when external pH was raised. At pH 7 or higher, the Cl- current fell during a hyperpolarizing voltage pulse and the rate of inactivation was directly proportional to the voltage change. The histidinemodifying reagent diethylpyrocarbonate (DEPC, 1 mM) which carbethoxylates histidil residues in proteins, suppressed the inactivation of Cl- currents at pH 7.5. On the other hand, no apparent changes in the kinetics of the currents at pH 5 were seen. No3- currents, which are independent of the extracellular pH and time, were not affected by DEPC. Our results support the notion that the inactivation of Cl- currents at pH 7.5 represents a membrane permeability change and that DEPC interferes with this process. Protonation of histidine groups associated with Cl- channels may be the controlling reaction for the pH -dependent Cl- response.

Animals↗
PubMed1996

Chloride currents in skeletal muscles of Bufo arenarum.

C1- currents (ICl) were measured in short fibers (1-2 mm) from the lumbricalis muscle of toads (Bufo arenarum) with two microelectrodes (15 degrees C). Initially the fibers were equilibrated in a high (K+)-containing solution: (mM) K2SO4 68; Na2SO4 20; KCl 60; CaSO4 8; MgSO4 1; HEPES 2.5. Constant pulses were applied when all the external K+ was replaced by Cs+: Cs2SO4 68; Na2SO4 20; CsCl 60; CaSO4 8; HEPES 2.5 (pH 7.5). Under these conditions about 80-90% of the current is carried by Cl-. The current-voltage relation is almost linear implying constant conductance and hence voltage-independent permeability. The voltage dependence of the net C1- current could be fitted by constant field equation with a PCl of 3.3 x 10-6 cm/sec. In a separate group of experiments a two-pulse technique was used to estimate the availability and the inactivation of the initial ICl during a test pulse. After returning the potential to the holding potential for various times, test pulses of the same amplitude and duration of the prepulses were applied. The initial current during the test pulse was 70% of the initial current during the prepulse and the recovery was complete in less than 300 msec with a linear relationship between the current during the test pulse and the amplitude of the preceding prepulse. When the test pulses were preceded by a positive prepulse, the initial current for any given test pulse was larger than with a negative prepulse. If we assumed that the initial current during the test pulse is a measure of the number of channels open at the end of the prepulse, these results suggest that hyperpolarizing pulses inactivate and depolarizing prepulses activate the ICl.

Animals↗
PubMed1996

The effect of aminophylline on the contraction threshold of rat diaphragm fibers and its modification by 9-aminoacridine.

We studied the effect of aminophylline (1mM) and 9-aminoacridine (100 microM) on the contraction threshold (CT) of rat diaphragm fibers (25 degrees C). The CT was measured by direct visualization (200 X) of the fiber under current-clamp conditions. The main findings are the following: 1) Aminophylline lowers the CT toward more negative values of the resting membrane potential (Vm). 2) 9-aminoacridine, a drug that diminishes Ca2+ release from the sarcoplasmic reticulum (SR), shifts the CT toward more positive values: 3) this effect is overcome by aminophylline. We suggest that the displacement in the CT to more negative Vm plays an important role in the potentiating effect of aminophylline. This could be the result of an enhancement of Ca2+ release from the SR.

Aminacrine↗
PubMed1995

[Ion channels in non excitable cells].

Several distinct types of voltage-gated and second-messenger-operated K+, Ca2+, Na+ and Cl- channels exist in electrically non excitable cells such as those of the hematopoietic lineage. In these cells ion channels mediate cellular functions involving intracellular biochemical responses, rather than rapid electrical signaling. The presence of the channels is required for several basic functions, such as activation, secretion of lymphokines, mitogenesis, the regulation of cell volume and the mechanisms of resistance to chemotherapeutic agents. Here IN we review the patch-clamp method for studying many characteristics of these ionic channels, particularly in blood cells.

Arachidonic Acid↗
PubMed1994

Frog striated muscle is permeable to hydroxide and buffer anions.

Hydroxide, bicarbonate and buffer anion permeabilities in semitendinosus muscle fibers of Rana pipiens were measured. In all experiments, the fibers were initially equilibrated in isotonic, high K2SO4 solutions at pHo = 7.2 buffered with phosphate. Two different methods were used to estimate permeabilities: (i) membrane potential changes were recorded in response to changes in external ion concentrations, and (ii) intracellular pH changes were recorded in response to changes in external concentrations of ions that alter intracellular pH. Constant field equations were used to calculate relative or absolute permeabilities. In the first method, to increase the size of the membrane potential change produced by a sudden change in anion entry, external K+ was replaced by Cs+ prior to changes of the anion under study. At constant external Cs+ activity, a hyperpolarization results from increasing external pH from 7.2 to 10.0 or higher, using either CAPS (3-[cyclohexylamino]-1-propanesulfonic acid) or CHES (2-[N-cyclohexylamino]-ethanesulfonic acid) as buffer. For each buffer, the protonated form is a zwitterion of zero net charge and the nonprotonated form is an anion. Using reported values of H+ permeability, calculations show that the reduction in [H+]o cannot account for the hyperpolarizations produced by alkaline solutions. Membrane hyperpolarization increases with increasing total external buffer concentration at constant external pH, and with increasing external pH at constant external buffer anion concentration. Taken together, these observations indicate that both OH- and buffer anions permeate the surface membrane. The following relative permeabilities were obtained at pHo = 10.0 +/- 0.3: (POH/PK) = 890 +/- 150, (PCAPS/PK) = 12 +/- 2, (PCHES/PK) = 5.3 +/- 0.9, and (PNO3/PK) = 4.7 +/- 0.5. PNO3/PK was independent of pHo up to 10.75. At pHo = 9.6, (PHCO3/PK) = 0.49 +/- 0.03; at pHo = 8.9, (PCl/PK) = 18 +/- 2 and at pHo = 7.1, (PHEPES/PK) = 20 +/- 2. In the second method, on increasing external pH from 7.2 to 10.0, using 2.5 mM CAPS (total buffer concentration), the internal pH increases linearly with time over the next 10 min. This alkalinization is due to the entry of OH- and the absorption of internal H+ by entering CAPS- anion. The rate of CAPS- entry was determined in experiments in which the external CAPS concentration was increased at constant external pH. Such increases invariably produced an increase in the rate of internal alkalinization, which was reversed when the CAPS concentration was reduced to its initial value.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1993

Effect of aminophylline-Ca2+ blocker interaction on membrane potential of rat diaphragm fibers.

We studied the antagonism between aminophylline and two calcium channel blockers, nifedipine and verapamil, and its effect on the resting membrane potential of rat diaphragm fibers in vitro at 25 degrees C. Aminophylline hyperpolarizes the fibers in a dose-dependent manner, and the maximum effect is reached with 1 mM of the drug, approximately 9 mV compared with normal values. Both nifedipine and verapamil (1-5 microM) decreased the amount of hyperpolarization induced by aminophylline, and this is partially reversed when the xanthine concentration in the bath is increased. From the Hill equation we obtained a value of 2 for the slope, suggesting that two molecules of aminophylline bind to the receptor. Nifedipine modifies the affinity and the intrinsic activity of aminophylline, whereas verapamil reduces its intrinsic activity. The effect of nifedipine and verapamil is explained on the basis of the changed action of aminophylline on its site as a result of the interaction of the calcium channel blockers with their interdependent receptors.

Aminophylline↗
PubMed1992

Blockers of potassium current and resting membrane potential in rat muscle fibers.

Rat diaphragm fibers were equilibrated for several hours in 150 mM KCl; when they were returned to 5 mM KCl the resting potential went back to its original level with a half time of 17 min. This repolarization was blocked by 5 mM BaCl2, a blocker of the inward rectifier K channel. On the other hand, 0.1 mM apamin and 0.02 mM glibenclamide which block the Ca-dependent and ATP sensitive K channels, respectively, and 0.1 mM 9-AC a blocker of the Cl- channel did not affect the repolarization. 5 mM barium decreased the K conductance measured under current-clamp conditions in diaphragm muscle fibers. The possible role of the inward rectifier system in the repolarization following return to normal [K]o is discussed.

Animals↗