[Study of cholinesterase activity in mixed saliva of patients with somatic pathology.]

Cholinesterase activity in mixed saliva of patients with somatic pathology was investigated. The results showed differences in enzyme activity depending on the disease. The highest salivary cholinesterase activity was detected in patients with bronchopulmonary pathology, and the lowest - in cardiovascular diseases. A reliable relationship between the changes in the activity of cholinesterase in saliva and the rate of salivation, and with patients taking atypical antipsychotics and M, H-anticholinergic drugs, was revealed.

[The high-pressure chemistry, barophysiological chemistry, comparative enzymology of cholinesterase the 100th anniversary from the birth of A. P. Brestkin].

There are exposed the main landmarks of the scientific biography of Professor Aleksandr Pavlovich Brestkin, connected with his investigations in the field of chemistry of high pressures, physiological chemistry of caisson disease, kinetics of esterase catalysis, and in comparative enzymology of cholinesterases.

[To the 80-anniversary of cholinesterase. The cholinesterase club in Sechenov Institute of Evolutionary Physiology and Biochemistry].

For the second half of the XX century, Sechenov Institute of Evolutionary Physiology and Biochemistry of the Russian Academy of Sciences was the center of the Russian cholinesterase investigations ("the Russian cholinesterase club"). The close cooperation with chemists-syntheticians of different scientific schools provided success and fruitfulness of this scientific search. All these years, there was preserved dualism of this investigation: a study of the mechanism of functioning and kinetics of cholinesterase catalysis as well as the comparative-enzymological character of studies of cholinesterases of the animals being at different levels of evolutionary development.

Scutellarin protects against Aß-induced learning and memory deficits in rats: involvement of nicotinic acetylcholine receptors and cholinesterase.

AIM: To examine the protective effects of scutellarin (Scu) on rats with learning and memory deficit induced by ß-amyloid peptide (Aß). METHODS: Fifty male Wistar rats were randomly divided into 5 groups: control, sham operation, Aß, Aß+Scu, and Aß+piracetam groups. Aß(25-35) was injected into the lateral ventricle (10 µg each side). Scu (10 mg/2 mL) or piracetam (10 mg/2 mL was intragastrically administered per day for 20 consecutive days following Aß treatment. Learning and memory was assessed with Morris water maze test. The protein and mRNA levels of nicotinic acetylcholine receptor (nAChR) α4, α7, and ß2 subunits in the brain were examined using Western blotting and real-time PCR, respectively. The activities of acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE) in the brain and plasma were measured using Ellman's colorimetric method. RESULTS: In Aß group, the escape latency period and first platform cross was significantly increased, and the total number of platform crossings was significantly decreased, as compared with the control and the sham operation groups. Both Scu and piracetam treatment significantly reduced the escape latency period and time to cross platform, and increased the number of platform crosses, but there were no significant differences between Aß+Scu and Aß+piracetam groups. In Aß group, the protein levels of nAChR α4 and α7 subunits in the cerebral cortex were significantly decreased by 42%-47% and 58%-61%, respectively, as compared to the control and the sham operation groups. Scu treatment caused upregulation of α4 and α7 subunit proteins by around 24% and 30%, respectively, as compared to Aß group, but there were no significant differences between Aß+Scu and Aß+piracetam groups. The protein level of nAChR ß2 subunit had no significant difference among different groups. The mRNA levels of nAChR α4, α7, and ß2 subunits were not significantly changed. In Aß group, the activities of AChE and BuChE in the brain were significantly increased, but were significantly decreased in the plasma, as compared to the control and the sham operation groups. Scu or piracetam treatment restored the activities in brain and plasma nearly to the levels in the control group. CONCLUSION: The results suggest that Scu may rescue some of the deleterious effects of Aß, possibly by stimulating nAChR protein translation and regulating cholinesterase activity.

[Update on the current role of plasma cholinesterase]. / Papel de las colinesterasas plasmáticas. Actualización.

The antagonism of steroidal nondepolarizing neuromuscular blockers (NDMBs) moved forward recently with the introduction of sugammadex, the only drug able to immediately reverse the effects of curarization produced by NDMBs. This advance has necessitated reflection on the future role of pseudocholinesterase. In spite of the side effects of succinylcholine and published opinions on its use, this NDMB continues to be used in clinical anesthesia. Pseudocholinesterase is mainly found in the liver, plasma, and nervous system. The enzyme is synthesized in the liver in greater amounts than required although certain conditions lead to deficiency, which is usually asymptomatic. The only clinical expression is the apnea which develops after administration of succinycholine because this NDMB cannot be metabolized. In some patients, slight reductions in the antagonism of succinylcholine lead to rising neuromuscular concentrations of the drug in accordance with the degree and duration of the blockade. We review the various forms of pseudocholinesterase deficiency, including a discussion of genetic variants, clinical manifestations, and management. In addition to discussing the diagnosis of this condition and the clinical implications, we highlight the importance of practice protocols and access to a referral laboratory if one is not available within the immediate hospital.

Computational studies on cholinesterases: Strengthening our understanding of the integration of structure, dynamics and function.

Computational approaches have proved valuable in elucidating structure/function relationships in the cholinesterases in the context of their unusual three-dimensional structure. In this review we survey several recent studies that have enhanced our understanding of how these enzymes function, and have utilized computational approaches both to modulate their activity and to improve the design of lead compounds for their inhibition. An animated Interactive 3D Complement (I3DC) is available in Proteopedia at http://proteopedia.org/w/Journal:Neuropharmacology:2.

Amyloid, cholinesterase, melatonin, and metals and their roles in aging and neurodegenerative diseases.

The aging brain shows selective neurochemical changes involving several neural cell populations. Increased brain metal levels have been associated with normal aging and a variety of diseases, including Alzheimer's disease (AD). Melatonin levels are decreased in aging, particularly in AD subjects. The loss of melatonin, which is synthesized by the pineal gland, together with the degeneration of cholinergic neurons of the basal forebrain and the deposition of aggregated proteins, such as the amyloid beta peptides (Abeta), are believed to contribute to the development of cognitive symptoms of dementia. Aging and its variants, such as AD, should be viewed as the result of multiple "hits," including alterations in the levels of Abeta, metals, cholinesterase enzymes, and neuronal gene expression. Herein, we present evidence in support of this theory, based on several studies. We discuss melatonin's neuroprotective function, which plays an important role in aging, prolongation of life span, and health in the aged individual. It interacts with metals and, in some cases, neutralizes their toxic effects. Dietary supplementation of melatonin restores its age-related loss. In mice, an elevated brain melatonin significantly reduced levels of potentially toxic Abeta peptides. Thus, compensation of melatonin loss in aging by dietary supplementation could well be beneficial in terms of reducing metal-induced toxicity, lipid peroxidation, and losses in cholinergic signaling. We propose that certain cholinesterase inhibitors and the NMDA partial antagonist memantine, which are FDA-approved drugs for AD and useful to boost central nervous system functioning, can be made more effective by their combination with melatonin or other neuroprotectants. Herein, we highlight studies elucidating the role of the amyloid pathway, metals, melatonin, and the cholinergic system in the context of aging and AD. Finally, melatonin is present in edible plants and walnuts, and consuming foodstuffs containing melatonin would be beneficial by enhancing the antioxidative capacity of the organisms.

Cholinesterase in embryonic development.

I. Cholinesterase (ChE) activity was studied histochemically during early development of the sea urchin, the amphibian, the chick and the rat embryo. After formalin fixation and embedding in water-soluble carbowax, the enzyme reaction was carried out in serial section. 2. Independent from innervation ChE appears in every embryonic blastema in a very early stage of development. It disappears from the embryonic cells after they have assembled into definite organ structures. Thus, ChE plays a role in embryonic development which is different from its known function in the adult. Therefore, ChE activity present in differentiating cells during a limited phase of development, is termed "embryonic cholinesterase". 3. Embryonic ChE was invariably found in cells engaged in morphogenetic movements. This observation has led us to suppose that the enzyme in involved in the regulation of cellular movements during development. 4. In particular, embryonic ChE is described in the following locations: a) During sea urchin gastrulation ChE is present in the primary mesenchymal cells emigrating from the blastula wall and in the archenteron cells which are known to bring about the invagination movement by contraction of their pseudopods. b) In the early chick blastoderm ChE active "droplet cells" are described which are supposed to emigrate from the epiblast layer in order to form the hypoblast. c) During development of notochord and somites, during closure of the neural tube and development of the head anlage, the close correlation of ChE activity with various morphogenetic movements is demonstrated: ChE appears during aggregation and desaggregation of epithelial compounds. The active bending of preexisting epithelial sheets, such as the neural plate, is also accompanied by ChE activity in epithelial cells...

Role of butyrylcholinesterase in canine tracheal smooth muscle function.

The role of butyrylcholinesterase (BuChE) and acetylcholinesterase (AChE) in in regulating acetylcholine (ACh) lifetime was investigated by use of selective cholinesterase (ChE) inhibitors. Addition of 1 microM tetraisopropylpyrophosphoramide (iso-OMPA) led to a 98% inhibition of BuChE activity with little or no effect on AChE activity. This inhibition was accompanied by a 26% increase in the amplitude and a 43% prolongation in the half-relaxation time of contractions elicited by electric field stimulation (EFS). Coapplication of BW 284C51 (a selective AChE inhibitor) and 1 microM iso-OMPA resulted in increases of 2-fold in the amplitude and 10-fold in the half-relaxation time of EFS-induced contractions. These alterations were accompanied by small but sustained baseline contractures that were antagonized completely by incubation with exogenous BuChE (2.5 U/ml). The results suggest that BuChE serves to coregulate the lifetime of ACh in canine tracheal smooth muscle.

Non-classical actions of cholinesterases: role in cellular differentiation, tumorigenesis and Alzheimer's disease.

The cholinesterases are members of the serine hydrolase family, which utilize a serine residue at the active site. Acetylcholinesterase (AChE) is distinguished from butyrylcholinesterase (BChE) by its greater specificity for hydrolysing acetylcholine. The function of AChE at cholinergic synapses is to terminate cholinergic neurotransmission. However, AChE is expressed in tissues that are not directly innervated by cholinergic nerves. AChE and BChE are found in several types of haematopoietic cells. Transient expression of AChE in the brain during embryogenesis suggests that AChE may function in the regulation of neurite outgrowth. Overexpression of cholinesterases has also been correlated with tumorigenesis and abnormal megakaryocytopoiesis. Acetylcholine has been shown to influence cell proliferation and neurite outgrowth through nicotinic and muscarinic receptor-mediated mechanisms and thus, that the expression of AChE and BChE at non-synaptic sites may be associated with a cholinergic function. However, structural homologies between cholinesterases and adhesion proteins indicate that cholinesterases could also function as cell-cell or cell-substrate adhesion molecules. Abnormal expression of AChE and BChE has been detected around the amyloid plaques and neurofibrillary tangles in the brains of patients with Alzheimer's disease. The function of the cholinesterases in these regions of the Alzheimer brain is unknown, but this function is probably unrelated to cholinergic neurotransmission. The presence of abnormal cholinesterase expression in the Alzheimer brain has implications for the pathogenesis of Alzheimer's disease and for therapeutic strategies using cholinesterase inhibitors.

Conclusions and comments. Xth International Symposium on Cholinergic Mechanisms.

Ancient medicine men of Egypt and Arabia employed, under another name, the cholinergic agents, as did the hunters, warriors and shamans of Africa and South America. An explosion of cholinergic science occurred in the last and the current century, and the ISCMs witnessed and catalyzed this progress. The Xth ISCM emphasized the molecular characteristics of the receptors, cholinesterase and of the system engaged in liberation of Ach.

Synaptic transmission between rat spinal cord explants and dissociated superior cervical ganglion neurons in tissue culture.

Physiological properties of the synapses formed between explants of spinal cord and dissociated autonomic ganglion neurons in tissue culture were studied using intracellular and extracellular stimulation and recording techniques (as well as iontophoresis) with a culture perfusion system allowing continuous microscopic observation during repeated changes of the bathing medium. The principal neurons of the superior cervical ganglion (SCGN) were dissociated from perinatal rats and the spinal cord explants were obtained from 15-day rat fetuses; these were allowed to mature for 3-10 weeks in co-culture. Recordings from over 1000 SCGN established that: (a) spontaneous small depolarizations and action potentials occurred in 20% of the SCGN studied, (b) the EPSPs observed in SCGN after spinal cord stimulation were sensitive to decreased Ca2+ and increased Mg2+, as well as to D-tubocurare, hexamethonium and mecamylamine, but not to atropine (at 10(-6) M concentration) or to the alpha-adrenergic blocking agents phentolamine or phenoxybenzamine; no potentiation of the EPSPs was seen with neostigmate or eserine, (c) acetylcholine directly applied to the SCGN was seen to mimic the responses seen after spinal cord stimulation; tetrodotoxin blocked both direct and iontophoretically fired action potentials, with only a suprathreshold acetylcholine potential remaining. These synapses were not sensitive to alpha-bungarotoxin. It is concluded that the synapses formed by spinal cord neurites on principal SCGN in tissue culture are nicotinic cholinergic, and that the evoked EPSPs recorded in this study are thus similar to the orthodromic fast EPSPs observed in vivo. No slow synaptic responses were observed and no demonstrable effects were noted that could be attributed to adrenergic transmission.

Are the effects of gestational stress on motor development and cerebellar cholinesterase activity mediated prenatally?

Prenatal random noise and light stress retards early motor development in the rat. To determine whether this delay is pre- or postnatally mediated and whether the developing cerebellum may play a role in the delay, rats were subjected to this form of stress and their offspring were fostered or cross-fostered onto control rats and vice versa. Their motor development was assessed by a series of behavioral tests between birth and day 10 of life. Prenatally stressed pups (Eo), whether reared by stressed (Em) or control mothers (Cm), showed a retardation in motor development. However, stressed pups matured earlier when reared by a control mother than by a stressed dam. Control pups (Co) raised by stressed mothers showed normal behavioral development. Cerebellar cholinesterase activity, which has been shown to undergo characteristic alterations with normal maturation, did not differ between stressed and control pups at birth. However, on day 10, cholinesterase activity was lower in EmCo than in CmCo pups, whereas that in CmEo and EmEo pups did not differ significantly from controls. It is concluded that three effects of random noise and light stress during pregnancy may be distinguished: (1) A prenatal effect as expressed by a delay in motor development, (2) a postnatal (rearing) effect on motor development and (3) a rearing effect expressed as a decrease in cerebellar ChE activity.

Clinical and analytical considerations in the utilization of cholinesterase measurements.

Many theories have been advanced but the true physiological function for serum cholinesterase has still not been identified. Evidence has been presented for the abnormal expression of cholinesterase genes in many types of human tumors. Cholinesterase measurements are still used to monitor exposure to organophosphate insecticides and their clinical application requires a good understanding of the inter and intra-individual variation, as well as some knowledge of the time sequence between exposure and measurement of the cholinesterase activity. The use of serum cholinesterase measurement in liver disease varies in different countries. A case has not been made for the cost-effectiveness of adding serum cholinesterase as part of a screening procedure for the diagnosis of liver disease. During the last 10 years much information has been obtained on the molecular biology and genetics of acetylcholinesterase and butyrylcholinesterase, distinct enzymes encoded by two different, but related genes. It has been established that BChE is included by a single gene which corresponds to the E1 locus. The complete amino acid sequence of human serum cholinesterase and the location of disulfide bonds within the sequence have been described. The molecular basis of many variants of human serum cholinesterase has been described in detail. It is not rare for multiple mutations to occur within a single butyrylcholinesterase gene or there may be combination of mutations. At least 11 silent variants of human butyrylcholinesterase have been identified. There still exists a wide variety of substrates and analytical conditions for butyrylcholinesterase measurement in a number of clinical situations. No real evidence has been provided for clinical value for their use in the diagnosis of Alzheimer disease or monitoring the use of cholinesterase inhibitors in the treatment of pre-senile dementia of Alzheimer type. However, the insights from molecular biology technology may well open up more challenges in a variety of clinical situations.

Eptastigmine augments basal and GHRH-stimulated growth hormone release in young and old dogs.

The aim of the present work was to evaluate the effect on the growth hormone (GH) secretion of eptastigmine, a new long-acting cholinesterase inhibitor, in unanesthetized beagle dogs. In a first study, 5 young dogs were given single doses (0.5, 1.0, and 2.0 mg/kg, i.m.) of the drug or saline in a randomized cross-over manner. Blood samples were collected immediately before and, at regular intervals, until 150 min after drug injection. GH plasma concentrations were determined by radioimmunoassay. Plasma cholinesterase activity was measured with a potentiometric method. There was a significant logistic relationship (r = 0.601, P < 0.01) between the administered dose of eptastigmine and the log-transformed areas under the GH plasma concentration-time curve (AUC) with a calculated ED50 for eptastigmine of 0.63 +/- 0.36 mg/kg. There was also a significant linear relationship (r = 0.630, P < 0.01) between log-transformed AUC of GH levels and AUC of plasma cholinesterase activity. In a second study we evaluate the ability of eptastigmine (2.0 mg/kg, i.m.) to potentiate the GH-releasing effect of the GH-releasing hormone (GHRH, 2.0 micrograms/kg, i.v.) in young and old dogs. Eptastigmine was administered 45 min before GHRH and blood collected every 15 min until 90 min after GHRH injection. In young dogs, maximum GH plasma levels (Cmax) were 6.1 +/- 1.0 ng/ml after GHRH compared to 22.5 +/- 2.3 ng/ml after GHRH preceded by eptastigmine (P < 0.01). In old animals, Cmax were 4.6 +/- 1.4 ng/ml after GHRH vs 13.2 +/- 7.4 ng/ml after combined administration of GHRH and eptastigmine (P < 0.05). These data indicate that eptastigmine is very effective in augmenting basal and stimulated GH secretion in old dog. The good activity also shown in old animals suggests a potential use of this drug to reverse the age-dependent decline in GH secretion responsible for many involutional changes of aging.

Non-quantal acetylcholine release after cholinesterase inhibition in vivo.

After anticholinesterase treatment in vivo, depolarization of the postsynaptic muscle fibre membrane by about 4 mV develops due to non-quantally released acetylcholine from the motor nerve terminal. This conclusion was supported by experiments with the curarization of diaphragm slices from anticholinesterase treated mice during intracellular microelectrode recordings.

Localization of a novel adhesion-promoting site on acetylcholinesterase using catalytic antiacetylcholinesterase antibodies displaying cholinesterase-like activity.

A monoclonal antibody (MAb) raised against human acetylcholinesterase was found to have catalytic activity. A similar phenomenon was observed in a polyclonal antibody raised against the same antigen. The antibodies were demonstrated to be pure, and no contamination with either acetylcholinesterase or butyrylcholinesterase was found. Both antibodies hydrolyzed acetylthiocholine, an acetylcholinesterase substrate, and the MAb followed Michaelis-Menten kinetics. Six other MAbs and one other polyclonal antibody showed no evidence of catalytic activity. Acetylcholinesterase is a key component in the transmission of the nerve impulse, and is also expressed nonsynaptically during embryonic development, and abnormalities in expression are seen in neural tumors and degenerative disorders. This unusual expression is believed to be associated with a novel function of the enzyme related to differentiation and cell adhesion. Autoantibodies to acetylcholinesterase have been observed in a variety of neurologic, muscular, and autoimmune disorders. In an investigation of the possible role of acetylcholinesterase in cell adhesion, we showed that the enzyme promoted neurite outgrowth in neuroblastoma cell lines, and conversely, that certain antiacetylcolinesterase antibodies abrogated cell-substrate adhesion. Interestingly, the antibodies most effective in this regard were catalytic. Preliminary epitope analysis indicated a conformational epitope in the N-terminal domain. This domain contains the active site within a deep gorge and the peripheral anionic site at the rim of the gorge. Peripheral-site inhibitors, but not active-site inhibitors, also interfered with adhesion, and competed with the catalytic monoclonal binding to acetylcholinesterase, indicating that the epitope recognized is associated with the peripheral anionic site. The inhibitor data also support the supposition that catalysis in these antibodies may have arisen from stable complexation of acetylcholinesterase with an inhibitor. We conclude that the catalytic antiacetylcholinesterase antibody interacts with structures associated with the peripheral anionic site, thus defining a novel site on the molecule involved in cell adhesion. This finding has implications for our understanding of the potential importance of this peripheral site in a variety of congenital, neoplastic, and degenerative conditions.

[Cholinesterases]. / Les cholinestérases.

OBJECTIVE: To review current data on butyrylcholinesterase. DATA SOURCES: Search through Medline data bases of articles in French or English. STUDY SELECTION: Original articles and case reports were selected. Letters to editor were excluded. DATA EXTRACTION: The articles were analyzed in order to obtain current data on biochemical structure, action, major pathological variations, especially with regard to the recent informations obtained by molecular biology concerning the identification of genetic variants. DATA SYNTHESIS: Butyrylcholinesterase must be differentiated from acetylcholinesterase, which cannot hydrolyse succinylcholine. The physiological action of butyrylcholinesterase remains unknown, although it can hydrolyse many drugs. Excluding genetical mutations, several physiopathological situations alter butyryl-cholinesterase activity. Butyrylcholinesterase activity assessment does not allow the diagnosis of genetic variants. Whatever the origin, only deficits of more than 50% modify significantly the metabolism of succinylcholine or mivacurium. The diagnosis of a prolonged neuromuscular blockade is obtained with systematic monitoring of the neuromuscular function in case of administration of mivacurium or succinylcholine. Mivacurium should only be re-injected when one response at train of four is obtained. In case of prolonged neuromuscular blockade, the anticholinesterasic agent should not be administered when no response at train of four is obtained. The biochemical methods using inhibitors (dibucaine, fluoride) of the butyrylcholinesterase and a familial study lead to the diagnosis in most cases because the atypical and fluoride variants are the most frequent. When results are doubtful, genetic molecular methods with the use of PCR and restriction enzymes allow a rapid diagnosis.

[Reorganization of the nervous system during regeneration in Dugesia tigrina]. / Perestroika nervnoi sistemy pri regeneratsii planarii Dugesia tigrina

The normal structure of the nervous system in Dugesia tigrina Girard and the total morphodynamics of the nervous system during regeneration have been studied by means of choline esterase assay. The nervous system reacts to local damages of the planarian body; accumulations of nervous elements form in the wound region. Following the transverse cut of a planarian, the regeneration of the nervous system is not reduced to the completion of lacking parts. In this case (as well as in that of asexual reproduction) the nervous system manifests a considerable morphological lability and undergoes morphological rearrangements accompanied by the appearance of additional, frequently unpaired, nerve trunks. The data obtained are to be taken into account in neurobiological studies on planarians.

[Interest of the cholinesterase assay during organophosphate poisonings]. / Intérêt du dosage des cholinestérases dans le cadre des intoxications aux organophosphorés.

Cholinesterases are the main targets of organophosphorus compounds. The two enzymes present in the blood (butyrylcholinesterase, BChE; acetylcholinesterase, AChE) are biomarkers of their systemic toxicity. Activity of the plasma BChE is very often determined as it allows a rapid diagnostic of poisoning and is a marker of the persistence of the toxicant in the blood. The activity of the red blood cell AChE gives a better picture of the synaptic inhibition in the nervous system but the assay is less commonly available in routine laboratories. Better biomarker of the exposure, it allows a diagnosis of the severity of the poisoning and helps to assess the efficacy of oxime therapy. Besides the practical aspects of blood collection and sample processing, and the interpretation of the assays, this review stresses the complementarity of both enzyme assays and recalls their crucial interest for the confirmation of poisoning with an organophosphorus in a situation of war or terrorist attack and for the monitoring of occupational exposures.