Effects of phenytoin on acetylcholinesterase activity and cell protein in cultured chick embryonic skeletal muscle

1978 ◽  
Vol 56 (2) ◽  
pp. 287-293 ◽  
Author(s):  
C. Michael Cisson ◽  
Richard K. Entrikin ◽  
Barry W. Wilson
Keyword(s):  
Ache Activity ◽  
Creatine Phosphokinase ◽  
Direct Action ◽  
Acetylcholinesterase Activity ◽  
Lactic Dehydrogenase ◽  
Pectoral Muscle ◽  
Cell Protein ◽  
Leucine Incorporation ◽  
Chick Embryos

Cultured pectoral muscle from 11-day-old chick embryos was treated for 48 h with phenytoin (diphenylhydantoin, DPH) in concentrations ranging from 15 to 270 μg/ml on days 7–9 in vitro. Acetylcholinesterase (AChE, EC 3.1.1.7), creatine phosphokinase (CPK, EC 2.7.3.2), and lactic dehydrogenase (LDH, EC 1.1.1.27) activities, [3H]leucine incorporation into protein, and total protein of the cultures decreased in a dose-related manner with DPH concentrations of 30 μg/ml and greater. Total AChE activity and AChE activity released into the medium were specifically decreased with 15 μg DPH per millilitre. In cultures treated chronically with 15 μg DPH per millilitre on days 5–13 in vitro, total AChE activity and AChE activity released into the medium were 66.0 ± 13.2 and 64.7 ± 11.8% of untreated controls, respectively, but cellular AChE activity, cell protein, and [3H]leucine incorporation into protein were unaffected. The results indicate that DPH specifically decreases the total net synthesis of AChE activity by a direct action on cultured chick embryo muscle.

Verapamil-induced changes of muscle culture acetylcholinesterase

1979 ◽  
Vol 57 (12) ◽  
pp. 1421-1426 ◽  
Author(s):  
Gary T. Patterson ◽  
C. Michael Cisson ◽  
Richard K. Entrikin ◽  
Barry W. Wilson
Keyword(s):  
Ache Activity ◽  
Creatine Phosphokinase ◽  
Pectoral Muscle ◽  
Total Cell ◽  
Cell Protein ◽  
Leucine Incorporation ◽  
Chick Embryos ◽  
Total Cell Protein ◽  
Induced Changes ◽  
Increased Activity

Pectoral muscle of 11-day-old normal chick embryos was cultured for 7 days and then treated with verapamil for up to 48 h to test the hypothesis that verapamil affects acetylcholinesterase (ACHE) activity. Cellular ACHE activity increased, activity released into the medium decreased, and net activity decreased with increasing concentrations of verapamil up to 30 μg/mL. Neither protein synthesis nor total cell protein content changed with concentrations of 1–30 μg of verapamil/mL. One hundred micrograms of verapamil per millilitre was toxic to cultures. To determine the effect of verapamil on newly synthesized ACHE, 30 μg of it per millilitre was added to cells treated with diisopropylphosphorofluoridate (DFP). During recovery from DFP, the amount of newly synthesized ACHE released into the medium was increased threefold. Cellular leucine incorporation, total cell protein, and creatine phosphokinase activity were unaffected by the presence of verapamil during cellular recovery from DFP. The results indicate that verapamil has a direct effect on skeletal muscle cells by specifically decreasing net synthesis of ACHE and (or) inhibiting its release from the cell.

scholarly journals Intraperitoneal Exposure to Nano/Microparticles of Fullerene (C60) Increases Acetylcholinesterase Activity and Lipid Peroxidation in Adult Zebrafish (Danio rerio) Brain

2013 ◽  
Vol 2013 ◽  
pp. 1-11 ◽  
Author(s):  
Gonzalo Ogliari Dal Forno ◽  
Luiza Wilges Kist ◽  
Mariana Barbieri de Azevedo ◽  
Rachel Seemann Fritsch ◽  
Talita Carneiro Brandão Pereira ◽  
...  
Keyword(s):  
Lipid Peroxidation ◽  
Oxidative Damage ◽  
Ache Activity ◽  
Transcriptional Control ◽  
Direct Action ◽  
Acetylcholinesterase Activity ◽  
Biological Properties ◽  
Adult Zebrafish ◽  
Antioxidant Responses

Even though technologies involving nano/microparticles have great potential, it is crucial to determine possible toxicity of these technological products before extensive use. Fullerenes C60are nanomaterials with unique physicochemical and biological properties that are important for the development of many technological applications. The aim of this study was to evaluate the consequences of nonphotoexcited fullerene C60exposure in brain acetylcholinesterase expression and activity, antioxidant responses, and oxidative damage using adult zebrafish as an animal model. None of the doses tested (7.5, 15, and 30 mg/kg) altered AChE activity, antioxidant responses, and oxidative damage when zebrafish were exposed to nonphotoexcited C60nano/microparticles during 6 and 12 hours. However, adult zebrafish exposed to the 30 mg/kg dose for 24 hours have shown enhanced AChE activity and augmented lipid peroxidation (TBARS assays) in brain. In addition, the up-regulation of brain AChE activity was neither related to the transcriptional control (RT-qPCR analysis) nor to the direct action of nonphotoexcited C60nano/microparticles on the protein (in vitroresults) but probably involved a posttranscriptional or posttranslational modulation of this enzymatic activity. Taken together these findings provided further evidence of toxic effects on brain after C60exposure.

Effect of Ethanol on Erythrocyte Acetylcholinesterase Activity

1986 ◽  
Vol 23 (4) ◽  
pp. 458-462 ◽  
Author(s):  
Nuha A Haboubi ◽  
D I Thurnham
Keyword(s):  
Ache Activity ◽  
Acetylcholinesterase Activity ◽  
Inhibitory Effect ◽  
In Vitro Experiments ◽  
Previous Exposure ◽  
Organophosphorus Poisoning ◽  
The Mean ◽  
Erythrocyte Ache ◽  
Early Indication

Erythrocyte acetylcholinesterase (AchE) activity was measured in the blood of 36 alcoholic subjects and 41 healthy volunteers. The mean activity in the alcoholics was significantly lower than that in the control subjects. In vitro experiments showed that ethanol inhibited the AchE activity immediately and in proportion to the concentration of ethanol used. Incubation times up to 6 h did not increase the inhibition significantly. Incubation of normal red cells with ethanol for 15 h, followed by washing, showed also that AchE activity was inhibited by the previous exposure to ethanol and that washing did not reduce the inhibitory effect. The possibility is considered that depressed erythrocyte AchE activity may be an early indication of potential disturbances of the autonomic nervous system. The importance of reporting ethanol intake in patients with organophosphorus poisoning is stressed.

scholarly journals In vivo and in vitro effects of fructose on rat brain acetylcholinesterase activity: an ontogenetic study

2014 ◽  
Vol 86 (4) ◽  
pp. 1919-1926 ◽  
Author(s):  
CARINE A. GUIMARÃES ◽  
MAIRIS S. BIELLA ◽  
ABIGAIL LOPES ◽  
PEDRO F. DEROZA ◽  
MARIANA B. OLIVEIRA ◽  
...  
Keyword(s):  
Cerebral Cortex ◽  
Ache Activity ◽  
Young Patients ◽  
Brain Structures ◽  
Acetylcholinesterase Activity ◽  
Control Group ◽  
Inherited Disorders ◽  
Old Rats

Increased fructose concentrations are the biochemical hallmark of fructosemia, a group of inherited disorders on the metabolic pathway of this sugar. The main clinical findings observed in patients affected by fructosemia include neurological abnormalities with developmental delay, whose pathophysiology is still undefined. In the present work we investigated the in vitro and in vivo effects of fructose on acetylcholinesterase (AchE) activity in brain structures of developing rats. For the in vitro experiments, fructose was added at increasing concentrations to the incubation medium. It was observed that fructose provoked an inhibition of acetylcholinesterase activity in cerebral cortex of 30-day-old-rats, even at low concentrations (0.1 mM). For the in vivo experiments, rats were killed 1 h after a single fructose administration (5 µmol/g). Control group received the same volume of saline solution. We found that AchE activity was increased in cerebral cortex of 30- and 60-day-old rats receiving fructose administration. Finally, we observed that AchE activity was unaffected by acute fructose administration in cerebral cortex, striatum or hippocampus of 15- and 90-day-old rats. The present data suggest that a disruption in cholinergic homeostasis may be involved in the pathophysiology of brain damage observed in young patients affected by fructosemia.

The Effect of Galactose Metabolic Disorders on Rat Brain Acetylcholinesterase Activity

2000 ◽  
Vol 55 (9-10) ◽  
pp. 852-856 ◽  
Author(s):  
Stylianos Tsakiris ◽  
Kleopatra H. Schulpis
Keyword(s):  
Enzyme Activity ◽  
Ache Activity ◽  
Direct Action ◽  
Acetylcholinesterase Activity ◽  
Classical Galactosemia ◽  
Control Value ◽  
Brain Ache ◽  
Cholinergic Dysfunction ◽  
The Brain ◽  
Brain Acetylcholinesterase

Abstract To evaluate whether in classical galactosemia galactose (Gal), galactose-1-phosphate (Gal-1-P) and galactitol (Galtol) affect brain acetylcholinesterase (AChE) activity, various concentrations (1-16 mм) of these compounds were preincubated with brain homogenates of suckling rats as well as with pure eel Electroforus electricus AChE at 37 °C for 1 h. Initially, Galtol (up to 2.0 mм) increased (25%) AChE activity which decreased, thereafter, reaching the control value in high Galtol concentrations. Gal-1-P decreased gradually the enzyme activity reaching a plateau (38%), when incubated with 8-16 mM. However, when the usually found 2 mм of Galtol and 2 mм of Gal-1-P. concentrations in galactosemia were added in the incubation mixture simultaneously, brain AChE was stimulated (16%). Galtol or Gal-1-P modulated brain AChE as well as enzyme activity of E.electricus in the same way. Gal, Glucose (Glu) and glucose-1-phosphate (Glu-1-P) had no effect on AChE activity. It is suggested that Galtol as well as Gal-1-P can affect acetylcholine degradation acting directly on AChE molecule. Consequently the direct action of these substances on the enzyme might explain the brain cholinergic dysfunction in untreated galactosemia patients.

ATP and Protein Synthesis Assays to Evaluate the Toxicities of Preservatives in Vitro

1991 ◽  
Vol 19 (1) ◽  
pp. 60-67
Author(s):  
Corrado L. Galli ◽  
Barbara Viviani ◽  
Giampiero Casartelli ◽  
Marina Marinovich
Keyword(s):  
Protein Synthesis ◽  
Protein Content ◽  
Cellular Protein ◽  
Cell Protein ◽  
Leucine Incorporation ◽  
Maximal Effect ◽  
Atp Level ◽  
Cellular Protein Content

Cellular protein content, protein synthesis, ATP level and lactate dehydrogenase (LDH) activity, measured in a murine epidermal cell line (HEL/30), were used as the endpoints for determining the cytotoxicities of 17 antimicrobial chemicals. The relative toxicities of the test compounds were quantified by the determination of the concentrations inducing a 50% inhibition of [3H]-leucine incorporation into proteins (IC50), causing a 50% reduction of ATP level or final cell protein content or producing the maximal effect on LDH leakage (EC50) after 2 hours of treatment. The results indicate a good correlation between both the reduction of ATP level and inhibition of protein synthesis and the minimal inhibitory concentration (MIC) on different microorganisms, suggesting that ATP and protein synthesis assays could be useful as prescreening methods for testing the cytotoxicities of preservatives.

In vitro Influence of Phenylalanine on Acetylcholinesterase Activity and DNA

1998 ◽  
Vol 53 (3-4) ◽  
pp. 291-293 ◽  
Author(s):  
Kleopatra H. Schulpis ◽  
George A. Karikas ◽  
Joana Tjamouranis
Keyword(s):  
Mental Retardation ◽  
Synaptic Transmission ◽  
Molecular Interaction ◽  
Ache Activity ◽  
Metabolic Disorders ◽  
Acetylcholinesterase Activity ◽  
Hplc Method ◽  
Molecular Effect ◽  
Permeability Changes

Abstract Acetylcholinesterase (AChE) is a significant compo­nent of the membrane contributing to the permeability changes during synaptic transmission and conduction. Phenylketonuria is a group of metabolic disorders in which phenylalanine (Phe) is highly elevated in blood (up to 0 .1 ᴍ) resulting in*mental retardation etc. AChE activity was measured spectrophotometrically after incu­ bation with various Phe concentrations. Phe interaction with DNA was evaluated with an established HPLC method. Phe was found to inhibit AChE almost 40% , at a concentration of 5 mᴍ, whereas a 62.5% DNA peak exclusion (molecular interaction) was observed when Phe was incubated with DNA at a concentration of 3 mᴍ. In addition the ratio of DNA: Phe determined the potency of the observed molecular effect.

scholarly journals Antimicrobial Agent Triclosan Inhibits Acetylcholinesterase Activity in Vitro in a Dose-Dependent Manner

2021 ◽  
Author(s):  
Narasimha Pullaguri ◽  
Andrea Kagoo ◽  
Anamika Bhargava
Keyword(s):  
Antimicrobial Agent ◽  
Ache Activity ◽  
Acetylcholinesterase Activity ◽  
Consumer Products ◽  
In Vivo Studies ◽  
Dependent Manner ◽  
Human Health Effects ◽  
Dose Dependent

The antimicrobial agent, Triclosan, is widely used in many consumer products. It has been designated as a "contaminant of emerging concern (CEC)" because its exposure is known to cause adverse ecological and human health effects. Triclosan is not labelled as GRAS/GRAE (generally recognized as safe and effective), but its use is still prevailing. In vivo studies have revealed that exposure to triclosan results in a decreased acetylcholinesterase (AChE) activity. However mechanistic insights into AChE inhibition by triclosan are missing. Using in vitro AChE activity assay with purified AChE, we show that triclosan acts as a direct inhibitor of AChE and inhibits AChE activity in a dose-dependent manner. Given the function of AChE, any alteration in its activity can be neurotoxic. Our results provide important mechanistic insights into triclosan induced neurotoxicity with AChE as a target.

In vitro Effects of Chlorpyrifos on the Acetylcholinesterase Activity of Euryhaline Fish, Oreochromis mossambicus

2010 ◽  
Vol 65 (3-4) ◽  
pp. 303-306 ◽  
Author(s):  
Janapala Venkateswara Rao ◽  
Pathakoti Kavitha
Keyword(s):  
Ache Activity ◽  
Kinetic Studies ◽  
Gill Tissue ◽  
Acetylcholinesterase Activity ◽  
Dependent Manner ◽  
Concentration Dependent Manner ◽  
In Vitro Effects ◽  
Brain Ache Activity ◽  
Vitro Effect

The in vitro effect of a widely used organophosphorus insecticide, chlorpyrifos (CPP), on the acetylcholinesterase (AChE) activity was studied in vitro. The kinetic constants Km and Vmax and the bimolecular constant ki were determined in vitro. The in vitro AChE study indicated that CPP is neurotoxic and that it alters the apparent Km values widely in a concentration-dependent manner, resulting in a competitive type of inhibition. Based on the ki values, the sensitivity of AChE in brain is greater than that in gill tissue, at 7.3 · 10 - 5 M and 11.92 · 10 - 5 M, respectively. The study points to the importance of kinetic studies and the results suggest that in biomonitoring programmes brain AChE activity can be a good diagnostic tool for CPP toxicity.

Alanine Reverses the Inhibitory Effect of Phenylalanine on Acetylcholinesterase Activity

2002 ◽  
Vol 57 (5-6) ◽  
pp. 506-511 ◽  
Author(s):  
Stylianos Tsakiris ◽  
Kleopatra H. Schulpis
Keyword(s):  
Ache Activity ◽  
Acetylcholinesterase Activity ◽  
Competitive Inhibitor ◽  
Inhibitory Effect ◽  
Electrophorus Electricus ◽  
Enzyme Stimulation ◽  
Brain Acetylcholinesterase ◽  
Competitive Activation

The aim of this work was to evaluate, in vitro, the effect of ʟ-alanine (Ala) on suckling rat brain acetylcholinesterase (AChE) and on eel Electrophorus electricus pure AChE inhibited by ʟ-phenylalanine (Phe) as well as to investigate whether Phe or Ala is a competitive inhibitor or an effector of the enzyme. AChE activity was determined in brain homogenates and in the pure enzyme after 1 h preincubation with 1.2 mm of Phe or Ala as well as with Phe plus Ala. The activity of the pure AChE was also determined using as a substrate different amounts of acetylthiocholine. Ala reversed completely the inhibited AChE by Phe (18-20% in 500-600 μᴍ substrate, p<0.01). Lineweaver-Burk plots showed that Vmax remained unchanged. However, KM was found increased with Phe (150%, p<0.001), decreased with Ala alone (50%, p<0.001) and unaltered with Phe plus Ala. It is suggested that: a) Phe presents a competitive inhibitory action with the substrate whereas Ala a competitive activation; b) Ala competition with Phe might unbind the latter from AChE molecule inducing the enzyme stimulation; c) Ala might reverse the inhibitory effect of Phe on brain AChE in phenylketonuric patients, if these results are extended into the in vivo reality.

Sign in / Sign up

Export Citation Format

Share Document