Hormetic response of cholinesterase from Daphnia magna in chronic exposure to triazophos and chlorpyrifos.

In vivo activity of cholinesterase (ChE) in Daphnia magna was measured at different time points during 21-day exposure to triazophos and chlorpyrifos ranging from 0.05 to 2.50 microg/L and 0.01 to 2.00 microg/L, respectively. For exposure to triazophos, ChE was induced up to 176.5% at 1.5 microg/L and day 10 when measured by acetylthiocholine (ATCh), whereas it was induced up to 174.2% at 0.5 microg/L and day 10 when measured by butyrylthiocholine (BTCh). For exposure to chlorpyrifos, ChE was induced up to 134.0% and 160.5% when measured by ATCh and BTCh, respectively, with both maximal inductions detected at 0.1 microg/L and day 8. Obvious induction in terms of ChE activity was also detected in daphnia removed from exposures 24 hr after their birth and kept in a recovery culture for 21 days. Results indicated that the enzyme displayed symptoms of hormesis, a characteristic featured by conversion from low-dose stimulation to high-dose inhibition. In spite of that, no promotion in terms of reproduction rate and body size was detected at any tested concentrations regardless of whether the daphnia were collected at end of the 21-day exposure or at end of a 21-day recovery culture. This suggested that induction of ChE caused by anticholinesterases had nothing to do with the prosperity of the daphnia population.

Old and new questions about cholinesterases.

Cholinesterases have been intensively studied for a long time, but still offer many fascinating and fundamental questions regarding their evolution, activity, biosynthesis, folding, post-translational modifications, association with structural proteins (ColQ, PRiMA and maybe others), export or degradation. They constitute an excellent model to study these processes, particularly because of the sensitivity and specificity of enzymic assays. In addition, a number of provocative ideas concerning their proposed non-conventional, or non-catalytic functions deserve to be further documented.

Rifabutin autoinduction is caused by involvement of cytochrome P450 and cholinesterase.

Rifabutin exhibits remarkable autoinduction of its elimination, but the mechanism behind it was not fully known. Our work showed that rifabutin administration increased the metabolism of rifabutin itself in both in vivo studies and liver perfusion, and the half-life was decreased by 71.08% and 12.74%, respectively. Further results showed that rifabutin administration increased CYP3A, CYP2D and cholinesterase levels by 87.2%, 57.3% and 65.14%, respectively. The autoinduction phenomenon of rifabutin may, therefore, be attributed to induction of cholinesterase and CYP450 isoenzymes, such as

Cholinesterase activity in human pulmonary arteries and veins: correlation with mRNA levels.

Isolated intact human pulmonary arteries and veins were used to determine the acetylcholinesterase (AChE) or butyrylcholinesterase (BChE) activities in the absence or presence of two selective cholinesterase (ChE) inhibitors, iso-OMPA or BW284c51, respectively. These results were compared with the mRNA levels for each enzyme in human pulmonary vessels. Total ChE activities measured in presence of acetylthiocholine (ACTI, 1 mM) in intact vascular preparations were 45+/-04 and 114+/-07 mU/g tissue in human pulmonary arteries (n=14) and veins (n=14), respectively. These activities were completely abolished in presence of 10 microM neostigmine. In both types of vessels AChE and BChE activities were observed. These activities were at least 2-fold higher in human pulmonary veins when compared with arteries and were correlated with the accumulation of the corresponding transcripts (n=8). In each type of vessel, similar total ChE activities were detected in homogenized and intact preparations, while in human bronchial preparations this activity was 5-fold higher in homogenates than in intact preparations. Together these results provide evidence that the ChE activities in human pulmonary vessels may be extracellular and that the higher activity measured in veins as compared to arteries was associated with the differential accumulation of the corresponding transcripts.

Sodium selenite supplementation during pregnancy and lactation promotes anxiolysis and improves mnemonic performance in wistar rats' offspring.

Selenium is a micronutrient which is part of selenoprotein molecules and participates in a vast number of physiological roles and, among them,we have fetal and neonatal development. Therefore, the aimof this studywas to evaluate possible behavioral changes in offspring of female rats supplemented during pregnancy and lactation with sodium selenite. To address that, we treated two groups of female rats by saline or sodium selenite at a dose of 1mg/kg through oral route and performed neurochemical and behavioral tests. In the offspring, the thyroid profile and hippocampal neurochemistrywere evaluated. Behavioral testswere performed in pups both during childhood and adulthood. We found out that selenium (Se) supplementation increased serum levels of triiodothyronine (25%, p b 0.001) and thyroxine (18%, p b 0.05) and promoted a tryptophan hydroxylase 2 (TPH 2) expression decrease (17%, p b 0.01) and tyrosine hydroxylase (TH) expression increase (202%, p b 0.01) in the hippocampus. The cholinesterase activity was decreased (28%, p b 0.01) in Se supplemented rats, suggesting a neurochemical modulation in the hippocampal activity. During childhood, the Sesupplemented offspring had a reduction in anxiety-like behavior both in elevated plus maze test and in light­dark box test. In adulthood, Se-treated pups had an increase in the locomotor activity (36%, p b 0.05) and in rearing episodes (77%, p b 0.001) in the open field test, while in the elevated plus maze test they also exhibited an increase in the time spent in the open arms (243%, p b 0.01). For the object recognition test, Se-treated offspring showed increase in the absolute (230.16%, p b 0.05) and relative index discrimination (234%, p b 0.05). These results demonstrate that maternal supplementation by sodium selenite promoted psychobiological changes both during childhood and adulthood. Therefore, the behavioral profile observed possibly can be explained by neurochemical changes induced by thyroid hormones during the critical period of the central nervous system ontogeny.

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...

Influence of chronic oral intake of cannabis extract on oxidative and hydrolytic metabolism of xenobiotics in rat.

Dietary intake of petroleum ether extract of cannabis leaves by rats in doses of 158, 250 and 500 mg/kg in the first, second and third week, respectively, caused selective induction of hepatic microsomal carboxylesterases/amidases without affecting the renal hydrolytic activity. Acetanilide N-deacetylase, p-nitrophenylacetate (NPA) esterase and acetylsalicylic acid (ASA) esterase I and II (active at pH 5.5 and 7.4) were stimulated 125, 64, 82 and 60%, respectively, whereas the activities of procaine esterase and acetylaminofluorene (AAF) N-deacetylase remained unaltered. The hydrolysis of acetylcholine was also unchanged. Upon withdrawal of treatment microsomal hydrolytic activity receded to basal levels within 7 days. Curiously though, the two-fold induction of thiacetazone N-deacetylase (118%), a cytosolic hydrolase, remained largely undiminished (62%). An appraisal of the hepatic cytochrome P450 mediated oxidative metabolism revealed approximately three-fold induction of aromatic hydrocarbon hydroxylase (AHH) metabolizing benzo(a)pyrene whereas the N-demethylation of aminopyrene was unaffected. These activities were restored to normal when resin administration was discontinued.

cDNA cloning and characterization of Geotrichum candidum lipase II.

Geotrichum candidum produces two extracellular lipases, I and II. A lipase II cDNA clone was isolated from a cDNA library by colony hybridization using the 32P-labeled fragment of lipase I cDNA isolated previously. The nucleotide sequence of lipase II cDNA determined by the dideoxy chain terminating method includes the N- and C-terminal amino acid sequences of lipase II, and the overall amino acid composition deduced from the cDNA coincides with that deduced on amino acid analysis of this protein. The cloned lipase II cDNA codes a protein of 544 amino acids and a part of the signal sequence of 13 amino acids. The peptide chain lengths of lipases I and II are the same, their overall identity being 84%. Furthermore, four Cys residues are completely conserved, which may participate in the formation of disulfide bridges. A homology search indicated that the G. candidum lipases and Candida cyclindracea lipase are homologous enzymes and that they are members of the cholinesterase family.

Possible induction of cholinesterase in epileptic patients treated with anticonvulsant drugs: relationship with lipoprotein levels.

The effect of enzyme-inducing anticonvulsant drugs on the serum concentrations of lipoproteins has been widely studied. However, there is little agreement between the results with regard to the possible development of a lipoprotein profile related to an increased or decreased cardiovascular risk. It has been suggested that cholinesterase (ChE) could be induced by these drugs, something of undeniable interest as ChE appears to have a relation to the metabolism of lipoproteins. The serum activity of ChE was determined in a group of 90 adult epileptic patients (56 male and 34 female) treated with phenobarbital, phenytoin, and carbamazepine. The liver enzyme induction produced by these drugs was then evaluated by determining serum gamma-glutamyltranspherase activity and urinary excretion of D-glucaric acid. A significant increase of serum ChE (p < 0.005) was found in the group of patients compared to a control group (n = 49) with a similar distribution for age and sex. A significant correlation was found for both male and female patients between ChE and concentrations of triglycerides, phospholipids, cholesterol, low-density lipoprotein (LDL) phospholipids, LDL-cholesterol, and apolipoprotein B (p < 0.01). Similarly, in female patients, ChE had a significant correlation with the total cholesterol/high-density lipoprotein (HDL) cholesterol and LDL-cholesterol/HDL-cholesterol ratios (p < 0.01). The ChE/HDL-cholesterol relationship, which has been proposed as a marker for cardiovascular risk, presented significant correlations with the total cholesterol/HDL-cholesterol and LDL-cholesterol/HDL-cholesterol ratios in patients of both sexes (p < 0.001). In the case of epileptic patients treated with enzyme-inducing anticonvulsant drugs, there may be an association between the possible induction of ChE and the metabolism of lipoproteins containing apolipoprotein B.

Ultrastructural localization of non-specific cholinesterase activity in rat muscle spindles.

Electron microscopical localization of non-specific cholinesterase activity was studied in the encapsulated part of rat hindlimb muscle spindles. After incubation of the muscle tissue in a medium containing butyrylthiocholine bromide as substrate and BW284c51 as the specific inhibitor of acetylcholinesterase, a distinct electron-dense precipitate corresponding to non-specific cholinesterase activity was found along the whole length of muscle spindles. The richest source of non-specific cholinesterase activity were the motor end-plates present in the polar and juxtaequatorial regions. Much smaller amounts of reaction deposits were found at the secondary sensory terminals in the juxtaequatorial zones. The primary sensory terminals in the equatorial zone contained only low amounts of the reaction product. A fine homogeneous reaction product was localized in the narrow spaces between Schwann cell processes or in gaps between the Schwann cell, and axonal and muscle membranes. A granular precipitate was localized on the basal lamina in the synapse region of motor terminals or covering Schwann cell processes and sensory terminals with adjacent intrafusal muscle fibres. Our results suggest that most of non-specific cholinesterase in muscle spindles is synthesized by the Schwann cells; but a small amount can also be synthesized by fibroblast-like cells forming the inner capsule of muscle spindles. Non-specific cholinesterase thus coexists with acetylcholinesterase at motor end-plates, but is single at sensory terminals. The function of non-specific cholinesterase in sensory receptors is still not clear. It seems most probable that non-specific cholinesterase in muscle spindles may play a role in the maintenance of the external milieu around nerve endings, especially in the sensory region.

Ethnic differences in the frequency distribution of serum cholinesterase activity.

We studied the activity of the enzyme pseudocholinesterase (serum cholinesterase) and its sensitivity to inhibition by dibucaine and fluoride in 400 (200 Iranian and 200 Irish) healthy subjects. The results show Irish subjects have significantly higher serum cholinesterase activity than Iranian subjects (7.82 +/- 0.14 vs 5.22 +/- 0.09 u/ml, mean +/- SEM, p < 0.01). Furthermore, the percent of inhibition of enzyme activity by dibucaine (82.19 +/- 0.68 vs 69.29 +/- 0.68) and fluoride (79.90 +/- 70.13 +/- 0.62) was also significantly higher (p < 0.001) in Irish than in Iranian subjects. One subject (Iranian) with very low pseudocholinesterase activity and a dibucaine number below 20 (atypical) had a history of apnoea following succinylcholine (suxamethonium). These data indicate that the frequency of atypical and heterozygote genes for cholinesterase activity leading to prolonged apnoea is much higher in Iranian than Irish populations. This study emphasises the importance of ethnic pharmacology.

[Synthesis of multiple forms of brain cholinesterases]. / Sintez mnozhestvennykh form kholinésterazy mozga.

Highly purified multiple forms of brain cholinesterase were isolated by a new method. Eight forms were identified as acetyl cholinesterases, seven forms--as butyryl cholinesterases, four forms--as mixed acetyl- and butyryl cholinesterases and four forms--as aliesterases. Biosynthesis of multiple forms of brain cholinesterase was monitored by following incorporation of 14C-glycine into the purified fractions. Cats were killed at different periods (3 hrs, 3,6 and 9 days) after intoxication with isopropyl hydroxymethyl fluorophosphate. The restoration of the enzymatic activity after the inhibition did not correspond to the incorporation of the labelled amino acid into the enzyme protein. There was a transcient increase in the incorporation of the label after an increase in acetylcholine concentration.

[Induction of various enzymes in the rabbit uterus with estradiol]. / Induktsiia nekotorykh fermentov matki krolikov éstradiolom.

Effect of estradiol propionate on activity of seven enzymes from rabbit uterus was studied. Simultaneously with the known estrogen-induced enzymes, activity of some other enzymes from uterus cells (tyrosine transaminase, acetylcholinesterase, butyrylcholinesterase) was also studied. The hormone induced all the enzymes studied except of butyrylcholinesterase. After induction with the estrogen a new isoenzyme fraction was found in peroxidase: at the same time, content of isoenzymes of lactate dehydrogenase, tyrosine transaminase and acetylcholinesterase was increased.

[Codominant nature of alleles of locus E2 controlling synthesis of human blood serum cholinesterase]. / Kodominantnaia priroda allelei lokusa E2, kontroliruiushchego sintez kholinésterazy syvorotki krovi cheloveka.

The C5 component of ChE2 (locus E2) of human serum is not an autosomal dominant trait. This component is a part of the group of isozymes C5-10 which can be detected by polyacrylamide gel electrophoresis in acid medium (pH 4.8). The new electrophoretic data show that polymorphism of ChE2 is determined rather by a pair of autosomal codominant alleles called ChE2A and ChE2B. Serum of subjects with phenotypes ChE2AB (C5+) and ChE2BB (C5-) shows the same level of cholinesterase activity on the average.