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[The histochemical evidence of the cholinesterase in the integument by means of the copper ferro cyanide methode (author's transl)].

With the aid of the Ferrocyanidmethod from KARNOVSKI und ROOTS ChE-positiv structures in the skin of various laboratory animals and of man were located. Specific cholinesterase is found in verves, around apocrine sweat glands and around eccrine sweat glands of some species. In the stratum basale of the epidermis from the rat you can demonstrate a high concentration of acetylcholinesterase. Unspecific cholinesterase is detectable in nerves, in the blood vessels, in the lumen of apocrine sweat glands and sebaceous glands.

Acetylcholinesterase↗

[Changes in the acetylcholine concentration and acetylcholinesterase activity in the thyroid gland tissues and in serum cholinesterase activity in goiter].

A study was made of acetylcholine concentration and of the acetylcholinesterase activity in the thyroid gland tissue, as well as of the serum cholinesterase activity in the toxic goiter. This disease was accompanied by an elevation of the acetylcholine concentration and a fall of the eacetylcholinesterase activity. As to serum cholinesterase--it changed but little in this disease.

Acetylcholine↗

Interactions of cholinesterase inhibitors and corticosteroids with the hypnotic effect of benzodiazepines in mice.

Interactions of cholinesterase inhibitors or dexamethasone with the hypnotic effect of benzodiazepines appear to be strongly dependent on the dose of the cholinesterase inhibitor used and to a lesser extent on the dose of the hypnotic. The duration of the loss of righting reflex caused by the benzodiazepines, diazepam or chlorodiazepoxide, is markedly prolonged by 500 micrograms/kg, significantly antagonized both diazepam and chlorodiazepoxide. Relatively higher doses of physostigmine have no significant effect on either diazepam or chlorodiazepoxide. Dexamethasone, in relatively low doses of 250 and 500 micrograms/kg or 500 micrograms/kg respectively, significantly reverses the effect of diazepam and chlorodiazepoxide, while in relatively high doses of 1000 micrograms/kg the hormone significantly potentiates the hypnosis produced by the benzodiazepines. It is suggested that the interactions described may be the result of a modification of possible changes in cholinergic activity caused by the benzodiazepines. Although as yet no unequivocal explanation can be offered for the different effects, the observations described in this paper may explain the highly contradictory findings of other authors in both animal and human studies.

Adrenal Cortex Hormones↗

Activation and blockade of cardiac muscarinic receptors by endogenous acetylcholine and cholinesterase inhibitors.

Cholinesterase inhibitors are known to potentiate the effects of acetylcholine (ACh) and vagal stimulation on the myocardium. The studies presented here demonstrate that cholinesterase inhibitors (ChEI) also have activity in isolated atria in the absence of extrinsic cholinergic stimulation and that, depending on the ChEI, either indirect stimulation or direct blockade of cardiac muscarinic receptors can occur. Muscarinic agonists inhibit cyclic AMP formation in atria and the ChEIs physostigmine, neostigmine and echothiophate likewise produce a marked attenuation of isoproterenol-stimulated cyclic AMP accumulation The effect of physostigmine appears to result from muscarinic receptor activation by endogenous ACh as it is blocked by atropine. In contrast, the ChEI ambenonium does not stimulate but instead blocks muscarinic receptors coupled to cyclic AMP accumulation. Radioligand binding studies provide direct evidence that both ambenonium and demecarium are relatively potent muscarinic receptor antagonists, whereas physostigmine and other ChEI have little direct receptor activity. Physostigmine and ambenonium also have different effects on heart rate in vivo, the former potentiating and the latter apparently blocking vagal tone. The inhibition of cyclic AMP formation produced by physostigmine can be used as a measure of the concentration of endogenous ACh available at muscarinic receptor sites. Physostigmine blocks cyclic AMP formation in atria incubated in the absence of calcium or in the presence of tetrodotoxin, suggesting that endogenous ACh is spontaneously released in the absence of neuronal activity or depolarization-secretion coupling.

Acetylcholine↗

[Trial of cholinesterase reactivators as proserine antagonists].

HI-6 and TMB-4 were the most effective and safe of 7 cholinesterase reactivators tested as agents for the prophylaxis of proserine poisoning of male mice. The reactivator HI-6 strongly potentiated the prophylactic efficacy of a mixture of atropine and arpenal administered in the doses sufficient for the blockade of both the m- and h-cholinoreactive systems of mice. As demonstrated by experiments in vitro, HI-6 and TMB-4 did not reacivate proserine-inhibited cholinesterase. The natural anticholinesterase activity of HI-6 was negligible. Based on the correlation of the data obtained to the reported data indicating that HI-6 has a low ganglioblocking activity it is inferred that the direct effect on the receptor is of no importance for the potentiating effect. It is assumed that HI-6 modulates the cholinoreactive systems, which leads to a dramatic increase of the efficacy of cholinolytics.

Animals↗

Prenatal diagnosis of neural tube defects using the cholinesterases.

The usefulness of a cholinesterase assay and electrophoresis in the prenatal diagnosis of neural tube defects was investigated in amniotic fluids from 1,512 women. The assay used a alpha-naphthyl acetate as substrate and measured the combined activity of the enzymes acetylcholinesterase (AChE, E.C. 3.1.1.7) and cholinesterase (ChE, E.C. 3.1.1.8); the activity of both enzymes was raised in amniotic fluid from women carrying open NTDs. The alpha-naphthyl acetate assay distinguished fetuses with neural tube defects from normal fetuses more effectively than assays using acetylthiocholine as substate. A perfect score could be obtained on the sample tested when both enzyme assay and electrophoresis were done on those samples with activity greater than or equal to 4 mU. There was no correlation between gestational age between 13-21 weeks and activity of AChE + ChE (r = 0.03). The electrophoretic band of AChE activity proved to be a valuable diagnostic adjunct to both AFP or the AChE + ChE assay. A similar band or AChE activity was seen in adult brain and intestine but not in kidney, heart, liver, or lung, or in sera from women carrying normal or NTD fetuses.

Amniotic Fluid↗

Plasma cholinesterase activity and the duration of suxamethonium apnoea in children.

Plasma cholinesterase activity and the duration of suxamethonium apnoea were studied in 89 children and in 15 adults for comparison. Whereas the level of cholinesterase activity was not different in the two groups, children showed a significantly shorter duration of apnoea. There was poor correlation between the enzyme activity and duration of apnoea. The shorter duration of action of suxamethonium may be due to the larger volume of extracellular fluid in children.

Adult↗

Development of prestriate visual projections in the monkey and human fetal cerebrum revealed by transient cholinesterase staining.

Cholinesterase (ChE) staining was used to reveal the timing and pattern of development of afferents to the prestriate visual cortex (areas 18, 19, 20, and 21 of Brodmann) in a series of developing human and monkey fetal brains. This investigation was possible because the nucleus pulvinaris of the thalamus, the main source of subcortical projections to the prestriate cortex, displays positive reactivity after thiocholine incubation during the last three quarters of gestation, while neighboring thalamic nuclei that project to the adjacent neocortical areas are unstained. Staining of the pulvinar and its prestriate projections passes through six broad stages. Stage I begins in both species at the end of the first third of gestation. Positively stained fibers originate from the pulvinar and enter but do not extend beyond the hemispheric stalk. During stage II, pulvinar axons gradually invade the intermediate zone of the occipital lobe, and in stage III they reach the level of the subplate zone. In stage IV, which occurs around mid-gestation in both species, cholinesterase-positive fibers accumulate within the subplate zone subjacent to the developing prestriate cortex. During stage V, ChE-positive fibers penetrate the prospective prestriate cortex but do not yet form the alternating columnar pattern characteristic of pulvinar input to this area in the adults. Rather, ChE activity is concentrated in two continuous bands situated within prospective layers III-IV and VI; also a narrow band is visible in upper layer I. In stage V a clear histochemical border forms between prestriate and striate areas with ChE activity in prospective area 17 limited mostly to the superficial strata of layers I and II. This histochemical differentiation precedes the emergence of cytoarchitectonic landmarks. During stage VI, which begins in the last fifth of gestation in both species, the pulvinar become progressively less stainable and its projections can no longer be traced by ChE histochemistry.

Afferent Pathways↗

[Properties of the nervous tissue cholinesterase and carboxylesterase in the locust, Locusta migratoria].

It was shown that locust cholinesterase splits various thiocholine esters with different rate. Hydrolysis of p-NPA is due to the effect of carboxylesterase. The latter differs from cholinesterase by a low sensitivity to eserine and cation-containing organophosphorus inhibitor methylsulfomethylate-O-ethyl-S-(beta-ethylmercaptoethyl) methylthiophosphonate, as well as by higher sensitivity to triorthocresylphosphate. The results obtained are discussed in relation to possible differences of the active surface of the enzymes studied.

Acetates↗

Structural difference at the active site of dibucaine resistant variant of human plasma cholinesterase.

Human plasma cholinesterase from five different genotypes -- E1U E1U, E1U E1A, E1A E1A, E1U E1S, E1A E1S, and E1U E1U C5+ -- was purified 8,000 fold from serum by a two-step procedure involving chromatography on DEAE-cellulose and preparative disc electrophoresis. The esterases were labeled with diisopropyl-1, 3-C14-fluorophosphate (DFP) aminoethylated, and digested by trypsin. The trytic digests were subjected to high voltage electrophoresis, and the radioactive peptides were detected by radioautography. Comparison of the peptides revealed different electrophoretic mobilities of the usual and atypical (dibucaine resistant) plasma cholinesterase peptides. The results are consistent with a structural abnormality of the active center in the variant enzyme. No difference was observed an the esteratic site of the enzyme with C5 component.

Binding Sites↗

[Electrocorticograms and the histochemical effects in the rat accompanying the reactivation of cholinesterases after the effects of an organophosphorus inhibitor].

The administration of paraoxon, an organophosphate inhibitor of cholinesterases, elicits theta rhythms, fast rhythms and paroxystical alterations on the electrocorticogram of the Rat. The disappearance of these abnormalities of the cortical activity after administration of aromatic oximes seems to be in good correlation with the restoration of cerebral cholinesterase activity revealed by histochemical method.

Animals↗

Immunological comparison of the usual and atypical human serum cholinesterase phenotypes.

Antiserum prepared against highly purified usual human serum cholinesterase (the most common phenotype) cross-reacted identically with the atypical serum cholinesterase. The level of circulating atypical enzyme protein, determined immunologically, was about 30% lower when the enzyme came from an atypical rather than a usual phenotype, and the level of enzyme activity measured enzymatically at Vmax with either o-nitrophenylbutyrate or benzoylcholine as substrate showed approximately the same degree of reduction. The average specific activity (activity at Vmax per microgram of enzyme protein) in sera from 28 usual and 20 atypical individuals did not differ significantly. These findings suggest that the atypical enzyme not only has altered catalytic properties (Km) but also might be synthesized more slowly, or cleared in vivo more rapidly, than the usual enzyme.

Antigen-Antibody Complex↗

Interlaboratory study of the proficiency of cholinesterase phenotyping.

We report the results of a proficiency survey into the accuracy of cholinesterase (EC 3.1.1.8) phenotyping undertaken by 12 laboratories over a three-year period, during which 21 samples of nine different genotypes were analyzed. Specimens from patients who were homozygous for the atypical variant AA or from UA and AF heterozygotes were identified without difficulty, but the less-well-recognized combinations AJ and AK were detected by only two and three laboratories, respectively. Some degree of difficulty was also experienced in differentiating among UU, UF, and FS specimens. Clinically significant ascription errors occurred on 11 occasions. We recommend that detailed studies of cholinesterase status be undertaken only by laboratories experienced in the assay, those likely to be able to produce accurate results and offer appropriate advice.

Cholinesterases↗

[Effect of cholinesterase inhibitors on the electrical excitability of the membrane of frog muscle fiber].

The effects of reversible (galanthamine, eserine) and irreversible organophosphate (armine) cholinesterase inhibitors on excitable muscle fibre membrane of the frog involved elongation of the rise and the half-decay time of the AP (galanthamine and eserine), the critical level of depolarization being shifted to a more negative value. Armine decreased the rise and the half-decay time, the critical level of depolarization being shifted to the positive direction; subsequent rising of the threshold often caused blocking of neuro-muscular transmission during the first minutes of the armine action, pretreatment of preparations with galanthamine (eserine) preventing the threshold increase and armine blocking of transmission. Probably mechanism of these drugs action on the membrane channels is discussed. The negative shift of critical level seems to underlie the protective action of galanthamine and eserine in case of poisoning with irreversible cholinesterase inhibitors.

Action Potentials↗

Comparative study of cholinesterase in two snails Pila globosa and Lymnaea acuminata.

Enzymatic activity of acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE) was measured in the nervous and cardiac tissue of two snails Pila globosa and Lymnaea acuminata. Data on substrate specificity, enzyme kinetic constants and specific enzyme inhibitors indicated that, L. acuminata possessed both AChE and BuChE, while, P. globosa had only AChE. Pharmacological experiments carried out on isolated organs of the two snails indicated that isolated organs of P. globosa were sensitive to low doses of acetylcholine (ACh) and to relatively higher doses of butyrylcholine (BuCh). The responses elicited by ACh could be enhanced by eserine but that of BuCh could not be increased by ethopropazine which has been shown to be a specific inhibitor of BuChE in snails. Isolated hearts of L. acuminata on the other hand, were more sensitive to BuCh as compared to ACh. The BuCh effect was enhanced by ethopropazine while the effect of both ACh and BuCh increased by prior application of eserine. There is a correlation between the occurrence of the two types of cholinesterase (ChE) and the sensitivity of the isolated organs to a specific substrate of these enzymes suggesting a possible correlation between transmitters and cholinesterase system.

Acetylcholinesterase↗

Plasma cholinesterase activity in bank blood.

Plasma cholinesterase activity was periodically measured in bank blood. Of the initial plasma cholinesterase activity 87% was retained after 21 days of storage at 4 C in citrate-phosphate-dextrose solution. In addition, plasma frozen for 7 weeks at -70 C showed no decrease in enzyme activity. No difference could be demonstrated in the stability of the enzyme related to whether the blood was stored in citrate-phosphate-dextrose or in acid-citrate-dextrose solution. Bank blood transfusion prior to or during succinylcholine administration should not significantly alter the duration of action of succinylcholine, and would probably be effective in the management of prolonged apnea resulting from a deficiency of this enzyme.

Adult↗

[Effect of pH on the cholinesterase hydrolysis of different substrates].

The effect of pH on the kinetic constants of cholinesterase-catalyzed hydrolysis of isoamylacetate and acetylthiocholine was investigated. For the first substrate the rate-limiting step is the enzyme acetylation reaction, while the overall hydrolysis rate of the second substrate is limited by the deacetylation reaction. It was concluded that a basic group is involved in the deacetylation reaction and in the non-covalent binding step of both ionic and non-ionic substrates. An acidic group participates in the both reaction steps. It was suggested that protonation of the basic group leads to a conformational transition of the free enzyme and can also participate in the catalytic mechanism. The second basic group characterized by Ka3 can be the functional group of the enzyme anionic site. Taken together, the influence of pH on cholinesterase catalysis can be generalized by the overall reaction scheme.

Animals↗

Characterization of serum cholinesterase variants by kinetic analysis and isoelectric focusing.

Serum cholinesterase enzyme (E.C. 3.1.1.8) shows a considerable degree of genetically determined heterogeneity. In this communication we describe some kinetic properties of serum cholinesterase variants (Km and Vmax) from individuals predisposed to prolonged apneic periods after administration of the anesthetic, succinylcholine. Isoelectric focusing, in a narrow pH range (pH 4-6) of sera from normal and atypical phenotypes permitted the detection of differences in protein bands near the isoelectric pH of the enzyme.

Adolescent↗