Slow Cl Research Articles

Pharmacokinetics of ethopropazine in the rat after oral and intravenous administration.

The pharmacokinetics of the anticholinergic drug ethopropazine (ET) have been studied in the rat after intravenous (i.v.) and oral administration. After i.v. doses of 5 and 10 mg/kg ET HCl, mean +/- S.D. plasma AUC were 9836 +/- 2129 (n = 4 rats) and 13096 +/- 4186 ng h/mL (n = 5 rats), respectively. The t1/2 after 5 and 10 mg/kg i.v. doses were 17.9 +/- 3.3 and 20.9 +/- 6.0 h, respectively. The Cl and V(dss) after 5 mg/kg i.v. doses were 0.48 +/- 0.10 L/h/kg and 7.1 +/- 2.3 L/kg, respectively. Statistically significant differences were present between the 5 and 10 mg/kg dose levels in Cl and V(dss). Oral administration of 50 mg/kg ET HCl (n = 5 rats) yielded mean AUC of 2685 +/- 336 ng h/mL. Mean plasma C(max), t(max) and t1/2 after oral doses were 236 +/- 99 ng/mL, 2.2 +/- 1.4 h and 26.1 +/- 5.4 h, respectively. Less than 1% of the dose was recovered unchanged in urine and bile. Ethopropazine is extensively distributed in the rat, and has relatively slow Cl in relation to hepatic blood flow in the rat. The drug appears to be extensively metabolized in the rat, and nonlinearity is present between the 5 and the 10 mg/kg i.v. doses. The drug displayed poor bioavailability (< 5%) after oral administration.

Pharmacokinetics of ethopropazine in the rat after oral and intravenous administration

The pharmacokinetics of the anticholinergic drug ethopropazine (ET) have been studied in the rat after intravenous (iv) and oral administration. After iv doses of 5 and 10 mg/kg ET HCl, mean±S.D. plasma AUC were 9836±2129 (n=4 rats) and 13 096±4186 ng h/mL (n=5 rats), respectively. The t1/2 after 5 and 10 mg/kg iv doses were 17.9±3.3 and 20.9±6.0 h, respectively. The Cl and Vdss after 5 mg/kg iv doses were 0.48±0.10 L/h/kg and 7.1±2.3 L/kg, respectively. Statistically significant differences were present between the 5 and 10 mg/kg dose levels in Cl and Vdss. Oral administration of 50 mg/kg ET HCl (n=5 rats) yielded mean AUC of 2685±336 ng h/mL. Mean plasma Cmax, tmax and t1/2 after oral doses were 236±99 ng/mL, 2.2±1.4 h and 26.1±5.4 h, respectively. Less than 1% of the dose was recovered unchanged in urine and bile. Ethopropazine is extensively distributed in the rat, and has relatively slow Cl in relation to hepatic blood flow in the rat. The drug appears to be extensively metabolized in the rat, and nonlinearity is present between the 5 and the 10 mg/kg iv doses. The drug displayed poor bioavailability (<5%) after oral administration. Copyright © 1999 John Wiley & Sons, Ltd.

Photodissociation dynamics of carbonyl chloride fluoride and its implications for phosgene three body decay

The photodissociation dynamics of COFCl has been studied by monitoring Cl fragments by resonance enhanced multi-photon ionisation and time-of-flight techniques at dissociation wavelengths near 235 nm. The COFCl heat of formation and the dissociation energy for C–Cl bond fission were for the first time experimentally determined: ΔfH00 (COFCl)=-397±15 kJ mol-1 and D0(COF–Cl)=364±8 kJ mol-1. 35% of the available energy is channelled into FCO rotation and bending vibration. The remaining energy is released as product translation. Other than in phosgene, no spin–orbit state selective behaviour is observed. The observations agree with a decay mechanism within the COFCl molecular plane as well as with a fragmentation via out-of-plane movement of the departing Cl atom. The structural and electronic similarity of COFCl and COCl2 allows conclusions to be drawn on the COCl2 dissociation. The findings are evidence for a previously proposed decay mechanism for the asynchronous concerted three body decay of phosgene into CO+2Cl. The first bond cleavage produces ground and excited spin–orbit state Cl and Cl*, respectively, with large kinetic energy release, while breaking the second C–Cl bond exclusively generates slow ground state Cl atoms.

Primary and secondary processes in the 193 nm photodissociation of vinyl chloride

We have investigated the photodissociation of vinyl chloride (H2CCHCl) at 193 nm using the technique of photofragment translational spectroscopy. The experiments were performed at the Chemical Dynamics Beamline at the Advanced Light Source and used vacuum ultraviolet synchrotron radiation for product photoionization. We have observed five primary dissociation channels following an initial π*←π excitation. The majority of Cl atoms originate from an excited-state dissociation. The remaining dissociation channels are consistent with competition on the ground electronic state following internal conversion from the optically prepared state. These channels include atomic and molecular hydrogen elimination, HCl elimination, and a translationally slow Cl elimination channel. We have also identified and characterized two secondary decomposition channels: (1) the elimination of Cl from chlorovinyl radicals following the primary atomic hydrogen elimination channel, and (2) hydrogen atom elimination from vinyl radicals following the primary atomic Cl elimination. By measuring the truncation in the translational energy distribution for C2H2Cl products from primary atomic hydrogen elimination we deduce a barrier for the reverse reaction of Cl+acetylene of 11±2 kcal/mol. Since Cl is known to add rapidly to acetylene with no activation barrier, we conclude that H loss primarily forms the ClCCH2 isomer, and that the observed 11 kcal/mol barrier pertains to a concerted addition/rearrangement path to form the α-chlorovinyl radical. Finally, we report low-resolution photoionization spectra for the nascent vinyl radical and HCl photoproducts, in which redshifts in the ionization onsets can be related to the internal energy content.

Membrane currents in immature oocytes of the Rana perezi frog

Immature oocytes of the Rana perezi frog were studied electrophysiologically to see if some of the unusual ionic channels found in Xenopus oocytes were also expressed in these cells. Growing oocytes showed a fairly linear current/voltage relationship (from -200 to +60 mV), whereas fully grown cells had several voltage-dependent conductances. Depolarizing pulses elicited a potassium current blocked by tetraethylammonium (TEA) and two kinetically different Ca2+-dependent Cl- currents (ICl(Ca)), both sensitive to niflumic acid. ICl(Ca), which have not been previously observed in Rana immature oocytes, were also found in response to acetylcholine or rabbit serum superfusion or intracellular injection of Ca2+. In addition, three different Cl- currents were activated in these cells by hyperpolarization: (1) a transient inward current dependent on a critical intracellular Ca2+ concentration; (2) an inward rectifier Cl- current, which was Ca2+ independent; and (3) a high threshold (over -140 mV), slow Cl- current, blocked by several divalent cations, 4,4'-diisothiocyanatostilbene-2,2'-disulphonic acid (DIDS) and 4-acetamido-4-isothiocyanatostilbene-2, 2'-disulphonic acid (SITS). The presence of most of these infrequent currents in immature oocytes of several frogs and toads suggests that they are not merely the result of random genomic expression but a programmed decision, probably related to a definite functional role.

P2 purinoceptor‐activated inward currents in follicular oocytes of Xenopus laevis.

1. ATP, UTP and 2MeSATP (10 microM) evoked fast, transient (2-20 s duration) and slow, sustained (60-120 s duration) inward currents in 118 of 285 follicular Xenopus oocytes but failed to activate inward currents in defolliculated Xenopus oocytes (n = 136). 2. The potency order for nucleotides was (at 10 microM): UTP = 2MeSATP > 2ClATP = ATP > ADP > alpha, beta-methylene-ATP > adenosine > ITP, with beta, gamma-methylene-ATP, CTP and GTP inactive. This potency order is atypical for any single P2 purinoceptor subtype. 3. EC50 values (for fast and slow inward currents) were (microM): ATP, 5.6 +/- 2.1 and 6.7 +/- 3.0; UTP, 1.0 +/- 0.3 and 0.9 +/- 0.3; 2MeSATP, 0.6 +/- 0.2 and 9.2 +/- 7.4 (n = 3). Suramin inhibited fast (IC50: 65 +/- 20 microM; n = 7) and slow (IC50: 57 +/- 10 microM; n = 7) inward currents evoked by ATP. Suramin also inhibited slow inward currents produced by UTP but potentiated those caused by 2MeSATP. 4. Fast and slow inward currents evoked in response to ATP and UTP reversed at -12 +/- 3 mV (n = 3), a value close to Erev for Na(+)-permeable ion channels present in oocytes. These same currents were reduced significantly when extracellular Na+ was substituted with choline and Erev changed to -30 +/- 5 mV (n = 3). Inward currents evoked by ATP and UTP were inhibited in a dose-dependent manner by amiloride (10-100 microM). 5. Fast and slow inward currents caused by 2MeSATP reversed at -29 +/- 3 mV (n = 3), a value close to Erev for Cl- channels in oocytes. Fast and slow responses to 2MeSATP were inhibited in a dose-dependent manner by the Cl- channel blocker NPPB (10-100 microM) and were only reduced slightly by amiloride (100 microM). 6. We conclude that two P2 purinoceptor subtypes are present on follicular Xenopus oocytes. 2MeSATP activates fast and slow Cl- currents (Erev = -30 mV) via a suramin-insensitive P2Y purinoceptor, while UTP and ATP activate fast and slow Na(+)-dependent currents (Erev = -10 mV) via a suramin-sensitive P2U purinoceptor.

Effects of defolliculation on membrane current responses of Xenopus oocytes.

1. Catecholamines, adenosine, gonadotrophins, vasoactive intestinal peptide (VIP) and E-series prostaglandins all elicit K+ currents in follicle-enclosed Xenopus oocytes. Evidence suggests that cyclic nucleotides act as intracellular messengers in the activation of this K+ conductance. Muscarinic agonists and some divalent cations (e.g. Co2+, Mn2+, Ni2+ and Cd2+) elicit slow oscillatory Cl- currents, which are activated through hydrolysis of inositol phospholipids and mobilization of intracellular calcium by inositol phosphates. 2. We investigated whether these membrane current responses were generated in the oocyte itself or in enveloping follicular cells which are coupled to the oocyte by gap junctions. Oocytes were defolliculated, either enzymatically using collagenase, or by manual dissection combined with rolling over poly-L-lysine-coated slides. Removal of follicular cells was checked using scanning electron microscopy. Membrane current responses of defolliculated oocytes were compared with responses seen in follicle-enclosed oocytes taken from the same ovary. 3. The K+ responses evoked by all the various hormones/neurotransmitters were either drastically reduced (greater than 90%) or abolished by defolliculation. K+ currents generated by the adenylate cyclase activator forskolin and by intraoocyte injection of adenosine 3',5'-cyclic monophosphate (cyclic AMP), or guanosine 3',5'-cyclic monophosphate were similarly reduced in defolliculated oocytes. In contrast, oscillatory Cl- currents to acetylcholine and divalent cations were selectively preserved through defolliculation. 4. Injection of cyclic AMP (1-20 pmol) into defolliculated oocytes had little or no effect on oscillatory Cl- currents elicited by ACh. However, the calcium-dependent transient Cl- current, activated by depolarization of the oocyte membrane, was consistently potentiated (100-900%) by injections of cyclic AMP (1-10 pmol). 5. These experiments suggest that cyclic nucleotide-activated K+ currents arise essentially in follicular cells and are monitored within the oocyte through electrical coupling by gap junctions. Oscillatory Cl- responses evoked by ACh and divalent cations are produced largely or wholly in the oocyte itself.

Further characterization of the slow muscarinic responses inXenopus oocytes

In immature follicular oocytes of the frog Xenopus laevis, application of muscarinic agonists evokes a complex response consisting of a fast and a slow Cl currents (the dominant responses), Cl current fluctuations, and a less prominent slow K current. The characteristics of the slow ACh-evoked potassium current were studied using the two-electrode voltage clamp method, and compared to those of the ACh-evoked Cl currents. In experiments designed to study the K current response separately, without the interference of ACh-evoked Cl currents, the holding potential was set close or equal to Cl equilibrium potential (measured as the reversal potential of the ACh-evoked Cl current). The Cl current responses were studied in cells that had negligible K current response. The dose-response curve of the potassium response followed classical Michaelis-Menten kinetics. The dose-response characteristics of the slow ACh-evoked Cl current displayed a positive cooperativity of at least 3. In spite of this difference, kinetic analysis revealed that these two responses, as well as the fast Cl current response that was characterized earlier (Dascal and Landau 1982), had almost identical apparent equilibrium dissociation constants (0.29-0.39 microM), suggesting involvement of a single receptor class. Both K and Cl currents were reduced (to 32-56% of control) by millimolar concentrations of phosphodiesterase (PDE) inhibitors, theophylline and isobutylmethylxanthine. Elevation of extracellular Ca concentration from 1 to 10 mM doubled the K current; depletion of external Ca caused a partial inhibition of this response. The K current was potentiated by 0.1 microM 4-phorbol 12,13-dibutyrate (PDBu).(ABSTRACT TRUNCATED AT 250 WORDS)

Single chloride-selective channels active at resting membrane potentials in cultured rat skeletal muscle

The patch-clamp technique was used to characterize channels that could contribute to the resting Cl-conductance in the surface membrane of cultured rat skeletal muscle. Two Cl- -selective channels, in addition to the Cl- -selective channel of large conductance described previously (Blatz and Magleby, 1983), were observed. One of these channels had fast kinetics and a conductance of 45 +/- 1.8 pS (SE) in symmetrical 100 mM KCl. The other had slow kinetics and a conductance of 61 +/- 2.4 pS. The channel with fast kinetics typically closed within 1 ms after opening and flickered between the open and shut states. The channel with slow kinetics typically closed within 10 ms after opening and displayed less flickering. Both channels were active in excised patches of membrane held at potentials similar to resting membrane potentials in intact cells, and both were open a greater percentage of time with depolarization. Under conditions of high ion concentrations, both channels exhibited nonideal selectivity for Cl- over K+ with the permeability ratio PK/PCl of 0.15-0.2. Additional experiments on the fast Cl- channel indicated that its activity decreased with lowered pHi and that SO2-4 and CH3SO-4 were ineffective charge carriers. These findings, plus the observation that the fast Cl- channel was also active in membrane patches on intact cells, suggest that the fast Cl- channel provides a molecular basis for at least some of the resting Cl- conductance. The extent to which the slow Cl- channel contributes is less clear as it was typically active only after excised patches of membrane had been exposed to high concentrations of KCl at the inner membrane surface.

Desipramine pharmacokinetics in Chinese and Caucasian volunteers.

In order to better define the role of pharmacokinetic variation in reported cross-ethnic differences in dosing patterns of some psychoactive drugs, single dose kinetics of the tricyclic antidepressant desipramine (DMI) were studied in 14 Chinese and 16 Caucasian healthy volunteers. DMI and 2-OH-DMI concentrations were assayed with h.p.l.c. in serial plasma and 24 h urine samples over 5 days following an oral 100 mg dose of DMI. Mean total clearance of DMI ( CLDMI ) from plasma was significantly (P less than 0.05) higher in the Caucasians (123 +/- 57 l/h) than in the Chinese (73.5 +/- 38.8 l/h). There was no significant difference in the apparent clearance of DMI by hydroxylation, fraction of dose metabolized to the hydroxy metabolite, DMI t1/2 or plasma protein binding between the two groups. Trimodal distribution of CLDMI was found, with 4/14 (29%) Chinese demonstrating slow CL (less than 33 l/h) and 4/16 (25%) Caucasians rapid CL (greater than 195 l/h). Correcting CL values for the greater mean weight of the Caucasians did not alter the pattern of distribution. CLDMI did not correlate with body weight. Although environmental factors cannot be ruled out, these results are consistent with genetically based differences in hepatic metabolism, probably affecting pathways in addition to hydroxylation, and suggest that 30% of Orientals would be at risk for toxicity from routine doses of tricyclics or similarly metabolized drugs.

Relationship between H +, anion, and monovalent cation movements and Ca 2+ transport in sarcoplasmic reticulum: Further proof of a cation exchange mechanism for the Ca 2+-Mg 2+-ATPase pump

Two spectroscopic probes of free internal Ca 2+ were used to determine the influence of H + and anion permeation on the active transport of Ca 2+ by skeletal sarcoplasmic reticulum. The studies were carried out on a well-characterized Ca 2+-Mg 2+-ATPase-rich sarcoplasmic reticulum fraction. Studies of D. McKinley and G. Meissner (1977, FEBS Lett., 82, 47–50) show that this fraction consists of two populations of vesicles: type I which has an electrically active monovalent cation (M +) permeability and type II which lacks it. The present study distinguishes between electrically active (charge-carrying) and electrically silent (e.g., countertransport) mechanisms of ion permeation in the two vesicles and shows how the active transport of Ca 2+ is influenced by these permeabilities. The major results are as follows: (1) Both type I and II vesicles have an electrically active H + permeability. (2) Type I vesicles have electrically active anion (A −) permeabilities; type II vesicles do not. (3) At low concentrations of nonpenetrating buffers, ion imbalances across the membrane can create pH imbalances. This is due to the coupling of M + and A − movements with H + movements. Following a jump in KCl concentration internal acidification is observed in type I vesicles while internal alkalinization is observed in type II vesicles. These pH gradients are dissipated on a time scale of seconds and tens of minutes for type I and II vesicles, respectively. (4) Tris(hydroxymethyl)aminomethane (Tris) was shown to be effective in dissipating pH gradients in type II vesicles. A model is proposed whereby HCl is equilibrated across the membrane by a Tris-catalyzed transport cycle involving transport of an ion pair between Tris-H + and Cl − and return of the unprotonated form of the buffer. (5) The permeabilities of several physiological and nonphysiological anions were determined for type I and II vesicles. Electrically active permeability was demonstrated for Cl − and phosphate in type I vesicles. Type II vesicles lacked electrically active mechanisms for these two anions. Evidence is given for slow Cl − OH − exchange and for rapid Cl − HCO 3 − exchange in type II vesicles. Electrically silent phosphate influx probably occurs by H 2PO 4 − OH − exchange. (6) Under normal conditions the Ca 2+ uptake of type II vesicles is masked. It can be unmasked by addition of nigericin in the presence of Tris. The combination of ionophore and penetrating buffer render the type II vesicles KCl permeable, allowing the replenishment of internal K + during active transport. The results are analyzed and shown to be in agreement with the Ca 2+-Mg 2+-ATPase pump acting as a Ca 2+ K + exchanger. The results are shown to be in disagreement with electrogenic models of pump function.

Mass transfer rate in liquid anion exchange processes—I: Kinetic of the two-phase acid-base reaction in the system trilaurylamine-toluene-HCl-water

By using a stirred cell with a constant interfacial area the rate of the two phase acid-base reaction between aqueous HCl and trilaurylamine (TLA) dissolved in toluene has been studied. A reaction mechanism, where interfacial chemical reactions are the main rate determining processes, has been hypothized on the basis of both kinetic experiments as well through the knowledge of the interfacial properties of the system. The reactions are the following: H + + ( TLA) i⇌( TLAH +) i slow Cl − + ( TLAH +) i⇌( TLAHCl) i fast ( TLAHCl) i+ TLA ⇌ TLAHCl +(TLA) i slow where i and the bar indicate interfacial and organic phase species respectively. An expression for the overall rate of reaction has been derived and the rate constants evaluated. By means of an approximate model, coupling diffusion and chemical reactions together, it has been shown that a possible diffusional contribution to the overall rate can affect the numerical values of the rate constants while the type of dependence of the rate of extraction on the concentrations is the same. The proposed reaction mechanism has been shown to hold up to equilibrium.