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Octanol reduces end-plate channel lifetime.

1. Post-synaptic effects of n-octanol at concentrations of 0.1-1 mM were examined in toad sartorius muscles by use of extracellular and voltage-clamp techniques.2. Octanol depressed the amplitude and duration of miniature end-plate currents and hence depressed neuromuscular transmission.3. The decay of miniature end-plate currents remained exponential in octanol solutions even when the time constant of decay (tau(D)) was decreased by 80-90%.4. The lifetime of end-plate channels, obtained by analysis of acetylcholine noise, was also decreased by octanol. The average lifetime measured from noise spectra agreed reasonably well with the time constant of decay of miniature end-plate currents, both in control solution and in octanol solutions.5. Octanol caused a reduction in the conductance of end-plate channels. Single channel conductance was on average about 25 pS in control solution and 20 pS in octanol.6. In most cells the normal voltage sensitivity of the decay of miniature end-plate currents was retained in octanol solutions. The lifetime of end-plate channels measured from acetylcholine noise also remained voltage-sensitive in octanol solutions. In some experiments in which channel lifetime was exceptionally reduced the voltage sensitivity was less than normal.7. In octanol solutions, tau(D) was still very sensitive to temperature changes in most cells although in some the temperature sensitivity of tau(D) was clearly reduced. Changes in tau(D) with temperature could generally be fitted by the Arrhenius equation suggesting that a single step reaction controlled the decay of currents both in control and in octanol solutions. In some cells in which tau(D) became less than 0.3 ms, the relationship between tau(D) and temperature became inconsistent with the Arrhenius equation.8. As the decay of end-plate currents in octanol solutions remains exponential, and the voltage and temperature sensitivity can be unchanged even when tau(D) is significantly reduced, it seems likely that octanol decreases tau(D) by increasing the rate of the reaction which normally controls the lifetime of end-plate channels.

Animals

An examination of octanol and octanal metabolism to octanoic acid by horse liver alcohol dehydrogenase.

The kinetics of the horse liver alcohol dehydrogenase (alcohol: NAD+ oxidoreductase EC 1.1.1.1) catalyzed metabolism of octanol and octanal to octanoic acid have been examined. On incubation of octanol with horse liver alcohol dehydrogenase in the presence of NAD+, NADH as well as octanal and octanoic acid were seen as the initial products. However, on continued incubation, the octanal concentration progressively decreased to where only negligible quantities were present in the incubation after 10 min. The production of NADH was biphasic. An initial phase was followed in about 2 min with a slower but linear rate of NADH production. The production of octanoic acid was approximately linear throughout the 10 min incubation period. Since octanal is an intermediate in the oxidation of octanol to octanoic acid, the ability of semicarbazide to inhibit the metabolism of octanol to octanoic acid was examined. At a concentration of semicarbazide which was 63 times the concentration of octanol in the incubation media, the rate of formation of octanoic acid was inhibited by only 30%. The results of these experiments suggest that in the oxidation of octanol to octanoic acid a portion of the octanal formed from octanol is not released from the enzyme but, in the presence of NAD+, is oxidized to octanoic acid.

Alcohol Oxidoreductases

The interaction of n-octanol with black lipid bilayer membranes.

The electrical capacities of black lipid films formed from monoolein + n-hexadecane and monoolein + squalane (or squalene) solutions have been measured in the presence of various concentrations of n-octanol. In addition, partition coefficients for n-octanol between n-hexadecane and 0.1 M NaCl, dielectric constants for octanol-hexadecane mixtures and the interfacial tension of films and film-forming lipid solutions against the aqueous phases have been determined. It is concluded that in "solvent-free" bilayers the octanol is unlikely to have changed the bilayer thickness by more than about 1 A. The bilayer tension, on the other hand, increases appreciably in the presence of octanol.

Electrochemistry

Simple procedure for determining octanol--aqueous partition, distribution, and ionization coefficients by reversed-phase high-pressure liquid chromatography.

The described simple, accurate, and precise reversed-phase high-pressure liquid chromatographic procedure is in excellent agreement with 1-octanol shake-flask partition or distribution coefficients over a 3.5 log range. A chemically bonded octadecylsilane support is persilated and coated with 1-octanol. With 1-octanol-saturated buffers as mobile phases, a stable baseline (compared to 1-octanol adsorbed on silica) is obtained rapidly, and the log relative retention times are highly correlated with unit slope to log distribution or partition coefficients obtained from the classical shake-flask procedures. Only relatively basic, unhindered pyridines deviate, probably because of binding with residual silinol sites. In addition, if the apparent pKa or pKab of an ionizable compound lies within the pH operating range of the column support, the apparent pKa or pKab usually can be determined simultaneously with log P by measuring the log distribution coefficient at several pH values. The procedure gives rapid results, requires little material, and can tolerate impurities.

Chemistry, Pharmaceutical

Direct measurement of octanol-water partition coefficients by high-pressure liquid chromatography.

A technique is presented for the direct measurement of octanol-water partition coefficients by HPLC. The method involves running solutes in octanol-saturated water as the mobile phase against water-saturated octanol entrained on an inert support. Log P correlates linearly with log tc for a number of standards. The measurable range in log P (so far) is -0.3 to +3.7. A critical review of chromatographic methods in Hansch analysis is given.

Chemistry, Physical

Amino acid side-chain contributions to free energy of transfer of tripeptides from water to octanol.

The location of amino acids in soluble or membrane proteins is related to the hydrophobicity of the side chains. Amino acid hydrophobicity values are based upon the thermodynamics of transfer from an aqueous to a nonaqueous environment. However, for certain hydrophilic residues uncertainty exists on the appropriate hydrophobicity values. We have measured the octanol-water partition coefficients (Po/w) of tripeptides of the sequence N-14C-acetyl-Ala-X-Ala-NH-tButyl (AcAlaXAlaNHtButyl), where the central residue X was either Gly, Ala, Phe, Trp, Pro, His, Asp, or Glu. The Po/w for the tripeptides agreed reasonably well with values calculated by the fragment method of D. J. Abraham and A. J. Leo (Proteins Struct. Func. Gen. 2, 130-152, 1987). The log Po/w of the uncharged form was 1.6, 2.7, and 2.5 greater than the log Po/w of the ionized form for the His, Asp, and Glu peptide, respectively. The new data on the pH dependence of the ionizable side chains, His, Asp, and Glu, should result in better prediction of the partition coefficient of peptides as a function of pH. The thermodynamic parameters were determined from the temperature dependence of partitioning. In the temperature range studied (2 to 65 degrees C) the transfer of tripeptides from water to octanol was entropy governed except for the ionized peptides. A heat capacity term was necessary to account for the transfer of tripeptides containing non polar residues. The heat capacity change for transfer from water into octanol was -45, -73, -81, and -88 cal/mol K for Ala, Phe, Trp, and Pro peptides, respectively. Peptides containing Gly, His (pH 7.2), and the uncharged forms of Asp, Glu, and His did not show a significant change in heat capacity. The side-chain contribution of the central residue X (delta Gx) to the free energy of transfer was obtained from the difference between the free energy of transfer of the peptide containing the central residue X and the Gly peptide; delta Gx = delta G(AcAlaXAlaNHtButyl) - delta G(AcAlaGlyAlaNHtButyl). The relative order of hydrophobicity of the side chains correlated well with previous studies. However, a significant difference was found for the absolute hydrophobicity between the present study and experimental data on N-acetyl amino acid amide derivatives (J. Fauchere and V. Pliska, Eur. J. Med. Chem. 18(4), 369-375, 1983).(ABSTRACT TRUNCATED AT 400 WORDS)

Amino Acids, Branched-Chain

[Influence of dinitrophenol, octanol and toluene upon pH-dependence of ca-ATPase activity of heavy meromyosin].

It is found that dinitrophenol, octanol and toluene produce similar effects on pH-dependence of ATPase of myosin and heavy meromyosin (HMM), i.e. they decrease or remove the neutral suppression of ATPase activity. The appearance of pH-dependence curves is simplified and approaches the form, which is characteristic for the ionisation curve of one; in the last resort two groups, participating in the enzyme activity. The activity of HMM is higher and the zone of the neutral suppression is diminished at low ionic strength, the activation by the modifiers being observed at the significantly lesser degree. CaATPase activation by dinitrophenol, octanol and toluene is suggested to be of the same nature and is accounted for the masking of "the inhibiting" ionizable group of the enzyme with near to neutral pK. This masking may be the result of the conformational changes occuring at the deformation of hydrofobic regions. The ionization of "the activity inhibiting" group of the enzyme depends directly or indirectly on the concentration of potassium chloride and the increase of KCl concentration bring to the inhibition of ATPase activity.

Adenosine Triphosphatases

Solvent-dependent conformational system of hydroxyureas in octanol-water and in inhibition of ribonucleotide reductase.

The carbonyl nitrogen IR absorption region of selected hydroxyureas was examined in octanol. The spectra indicated appreciably different absorptions prior to and after equilibration with water. Stability studies on the hydroxyureas indicated that the absorption differences were not due to chemical decomposition in the equilibration process but were due to solvent conformation and/or tautomeric dependency of the system. Preliminary in vitro inhibition of the enzyme ribonucleoside diphosphate reductase by selected hydroxyureas also indicated an apparent solvent dependency, which may involve the conformational and/or tautomeric properties of these agents. The implications of the solvent-dependent conformation-tautomeric system observed are discussed in relation to this property in the biological action of these agents.

Chemical Phenomena

Application of SCAP to drug design. 1. Prediction of octanol-water partition coefficients using solvent-dependent conformational analyses.

The solvent-dependent conformational analysis procedure (SCAP) has been used to predict the octanol-water partition coefficients of 20 different compounds with an average absolute error of 9%. SCAP predicts partition coefficients almost as well as the Hansch procedure using pi constants where the absolute error for the 20 compounds is 5%. In addition to estimating partition coefficients, SCAP allows direct calculation of the corresponding solute-solvent interaction free energies. Moreover, binding free energies, based upon hydrophobic and polar interactions, may also be computed. Such free energies are not calculable using other available methods. SCAP also allows solvation free energies to be compared to, or analyzed with, the various intramolecular free energies of the solute molecule as well as all other associated conformational properties.

Energy Transfer

Correlation of psychotomimetic activity of phenethylamines and amphetamines with 1-octanol-water partition coefficients.

In an attempt to relate the hallucinogenic potencies in man of some biologically important amphetamines and phenethylamines, the 1-octanol-water partition coefficients for 11 amphetamines were determined. Using these values and published Hansch pi constants, the log P for 17 additional amines was estimated. It was found that lipophilicity, as measured by the log of the partition coefficient, may be a significant determinant of the level of hallucinogenic potency. The study also suggests that an ideal log P value for psychotomimetric activity in man may be from 2.89 to 3.72.

Amphetamines

Polymorphisms and inheritance patterns of tetrazolium oxidase and octanol dehydrogenase in the house fly.

Zymogram analyses suggested the existence of two autosomal codominant alleles each at loci controlling the production of tetrazolium oxidase (To1) and octanol dehydrogenase (Odh1) in Texas house flies; an additional variant of the To1 locus was detected in a population from South Dakota. The allozymes of both systems may be dimeric as indicated by the presence of possible heteropolymers in flies heterozygous for To1 and Odh1 allels. Apparent nonsegregating isozymes of To and Odh (To2 and Odh2) were also noted in the zymograms of Texas flies. Some banding forms in the Odh gels could not be interpreted genetically.

Alcohol Oxidoreductases

Genetic analysis of two enzyme polymorphisms in a malaria vector mosquito: octanol dehydrogenase and acid phosphatase in Anopheles culicifacies Giles.

Genetic analysis was performed on two polymorphic enzyme systems in the malaria vector, Anopheles culicifacies Giles. The data indicate that both enzymes, octanol dehydrogenase and acid phosphatase, are controlled by autosomal loci but that these two loci are not linked. The three expected linkage groups in this mosquito have now been identified: linkage group I contains sex and rose-eye; linkage group II contains Odh; and linkage group III contains Acph.

Acid Phosphatase

Relationship between steroid permeability across excised rabbit cornea and octanol-water partition coefficients.

Permeability rates were determined across excised rabbit corneas for 11 steroids. Permeability coefficients for each steroid were calculated, and their logarithms were plotted against their respective log octanol-water partition coefficients. A parabolic relationship resulted, with an optimum log permeability and coefficient observed at a log Po of 2.9. From these experimental results, an improvement in ophthalmic bioavailability of dexamethasone acetate as compared to dexamethasone is predicted and correlates with literature results.

Animals

The study of genic variation by electrophoretic and heat denaturation techniques at the octanol dehydrogenase locus in members of the Drosophila virilis group.

Heat denaturation studies of three different electrophoretic allozymes of octanol dehydrogenase were performed from 10 species of the Drosophila virilis group. A total of 18 alleles were discovered, and in 3 species in which electrophoretic studies indicated that the locus was monomorphic, heat denaturation showed that the locus was polymorphic. We show that electrophoretic studies underestimate the number of alleles at this locus by a factor of 2.6 in these species. The results are discussed in the light of the continuing controversy over selection and neutral theories of genetic variation.

Alcohol Oxidoreductases

[Activity of the octanol dehydrogenase, of the alcool dehydrogenase and aldehyde dehydrogenase on the farnesol metabolism. Photoperiodic and neurhormonale regulation, controlling the metabolism of the juvenile hormone, in Pieris brassicae (author's transl)].

The antagonistic photoperiodic behaviour of the farnesol dehydrogenases indicates that the photonic control mechanism of the brain acts on the farnesol derivates. This cerebral control is double. The first system, linked at the allatotrope function is proportionnal at the photoperiod and acts on the octanol dehydrogenase 0,32. The second system controle the deshydrogenases ADH bands 0,50--0,58, is linked at the darkness. It is linked also at the neurocerebral activity then it stops its activity at the 4th day of the 5th stage. This last seems to be the determinating control for the establishment of the diapause since in short photoperiod, when the inhibition by this system ends, the alcool dehydrogenases 0,50-0,58 series is suractivated in rate with the lasting of the scotophase. In darkness, the 1st system functionnes cyclically and has a maximum synchron with the single maximum of the 2nd system. Inversally, in continuous light, the 2nd system is synchronisated with the 1st which has a prolongated action, maybe linked with a prolongated activity of the neurosecretory cells of the pars intercerebralis and corpora allata.

Animals

Chloride current induced by alcohols in rat dorsal root ganglion neurons.

We have recently demonstrated that ethanol and longer-chain alcohols (n-alcohols) enhance gamma-aminobutyric acid (GABA)-induced chloride currents before desensitization takes place. The potencies of n-alcohols increase with lengthening of the carbon chain. We now report that n-alcohols induce chloride currents by themselves in rat dorsal root ganglion neurons in primary culture. The whole cell variation of the patch clamp techniques was used to record currents as induced by external application of alcohols and other test compounds. Ethanol, n-butanol, n-hexanol and n-octanol induced inward currents with their potencies increasing in that order. The potencies were approximately one order of magnitude less than those to augment GABA-induced currents. The maximum amplitudes of currents induced by the alcohols were less than those produced by GABA. The n-octanol-induced currents were carried largely by chloride ions because the reversal potentials were changed according to the Nernst chloride potential as the internal chloride concentration was changed. Bicuculline and picrotoxin suppressed the n-octanol-induced current, and chlordiazepoxide and pentobarbital augmented the n-octanol-induced current. Therefore, the alcohol-induced chloride currents flow through the chloride channels associated with the GABAA receptors. When applied after the GABA-induced current was desensitized to a lower level, n-octanol suppressed rather than augmented the current. Thus, n-alcohols mimic barbiturates in augmenting the GABA-induced currents and in generating chloride currents by themselves. These actions of both agents may play a role in causing anxiolytic, sedative and/or anesthetic effects.

Alcohols