Detachment of retina with use of diisopropyl fluorophosphate (fluropryl) in treatment of glaucoma.
Explore the source record for details and available documents.
Fluorides↗
SEARCH · PubMed Health
Results for “ISOFLUROPHATE”
Explore indexed PubMed citations for clinical trials, systematic reviews and public health research. Read source abstracts and follow each citation to its original PubMed record.
Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.
Cholinesterases and contractility of cat irides. Effect of echothiophate iodide.
Explore the source record for details and available documents.
Acetylcholine↗
Anticholinesterase induces nicotinic receptor modulation.
The effects of carbamate anticholinesterases, pyridostigmine and physostigmine, on the function of the nicotinic receptor (nAChR) in TE671 cells was studied, precluding their inhibition of acetylcholine hydrolysis by carbachol usage. In radioassay, the simultaneous application of carbachol and carbamates dose-dependently decreased carbachol-induced 22Na+ influx, compared with carbachol activation alone. Increasing cell preincubation in the presence of carbamates, however, potentiated influx at low concentrations in a time-dependent manner. This facilitating effect of carbamates, even at high concentrations, was significantly increased by washing out these drugs and was blocked by pretreatment with diisopropylfluorophosphate. Similar results were also obtained in whole-cell patch-clamp study. There were insignificant changes in desensitization properties during facilitation. It is thus supposed that facilitation cannot be explained by the inhibition of acetylcholine hydrolysis. These results support a previous hypothesis that acetylcholinesterase might modulate nAChR by an unknown mechanism. In addition, the clinical effects of carbamates may be partly attributed to this facilitation.
Carbachol↗
Diisopropylphosphorofluoridate-induced depression of compound action potential of frog sciatic nerve in vitro is mediated through the inhibition of cholinesterase activity.
Effect of diisopropylphosphorofluoridate (DFP), an irreversible cholinesterase (ChE) inhibitor, on compound action potential (CAP) of sciatic nerve in vitro was examined. Further, the role of cholinesterase reactivator (1 acetyl-4-hydroxy imino methyl pyridinium bromide; SPK-3) in reversing DFP-induced changes was also evaluated. Diisopropylphosphorofluoridate produced a dose-dependent depression of the CAP. A concentration as low as 0.01 microM DFP produced a 5% depression (P < 0.05) and the maximal depression (30% of control) was observed with 1 microM. The SPK-3 (up to 10 microM) had no effect on the CAP; SPK-3 (10 microM) antagonized the DFP-induced depression of the CAP partially but not after 1 microM DFP. However, the inhibitory concentration of DFP to produce 50% of the maximal depression (IC50) was 0.38 +/- 0.025 microM in the presence of SPK-3 (10 microM; n = 4), against 0.15 +/- 0.05 microM for DFP alone (n = 7). These IC50 values were significantly different (P < 0.05, Student's t-test). The DFP decreased nerve ChE activity by 41% in the absence of SPK-3 and by 31% in the presence of SPK-3. Although SPK-3 could not completely reactivate the inhibited enzyme, it seems reasonable to conclude that the DFP-induced depression of the action potential of sciatic nerve was mediated by inhibiting the ChE activity.
Action Potentials↗
Rapid, high-sensitivity imaging of radiolabeled gels with microchannel plate detectors.
A microchannel plate detector has been used to image tritium-labeled protein on one- and two-dimensional electrophoresis gels. The good spatial resolution (70 microns) and high sensitivity (6.0 dpm/mm2) of the imaging system allows detection of low levels (femto moles) of labelled proteins. We are currently using the detector for identification of new targets involved in organophosphate neurotoxicity.
Animals↗
Degradation of organophosphorous nerve agents by enzyme-polymer nanocomposites: efficient biocatalytic materials for personal protection and large-scale detoxification.
The biocatalytic destruction of organophosphates has become an important focus area, as efficient "clean" technologies are sought for chemical weapons decommissioning, counteracting nerve agent attacks, and protecting against organophosphate pesticide poisoning. A novel method is advanced for immobilizing the broad-spectrum enzyme organophosphorous hydrolase (OPH) from Pseudomonas diminuta, based on the formation of nanocomposite protein-silicone polymers. The resulting materials are highly active, stable, and versatile biocatalysts for the liquid and gas phase detoxification of organophosphates, and can be fabricated as monoliths, sheets, thick films, granulates, or macroporous foams. This approach offers an efficient avenue to robust, high-performance biocatalytic OPH-containing polymers that outperform immobilized OPH catalysts reported to date. The method provides for the first time a route to biocatalytic materials that may be suitable for "active" protective wear, as well as bulk catalysts for the destruction of large volumes of organophosphates. The preparation of OPH-silicone biocomposites, their performances in the liquid and gas phase detoxification of paraoxon, dichlorvos, and diisopropyl fluorophosphate, and their features are discussed.
Adsorption↗
Nanocrystalline metal oxides as destructive adsorbents for organophosphorus compounds at ambient temperatures.
Nanocrystals of magnesium oxide react with organophosphorus compounds at room temperature by dissociative chemisorption, which we term "destructive adsorption". This process involves cleavage of P-O and P-F bonds (but not P-C bonds) and immobilization of the resultant molecular fragments. These ultrafine powders have unusual crystalline shapes and possess high surface concentrations of reactive edge/corner and defect sites, and thereby display higher surface reactivity, normalized for surface area, than typical polycrystalline material. This high surface reactivity coupled with high surface area allows their use for effective decontamination of chemical warfare agents and related toxic substances. Herein data is presented for paraoxon, diisopropylfluorophosphate (DFP), and (CH3CH2O)2P(O)CH2-SC6H5 (DEPTMP). Solid-state NMR and IR spectroscopy indicate that all OR and F groups dissociate; this leaves bound -PO4, -F, and -OR groups for paraoxon, DFP, and DEPTMP, respectively. For paraoxon, it was shown that one monolayer reacts. For DEPTMP, the OR groups dissociate, but not the P-CH2SC6H5 group. The nanocrystalline MgO reacts much faster and in higher capacity than typical activated carbon samples, which physisorb but do not destructively adsorb these phosphorous compounds.
Adsorption↗
Purification and characterization of hemolymph juvenile hormone esterase from the cricket, Gryllus assimilis.
Juvenile hormone esterase (JHE) from the serum of the cricket, Gryllus assimilis, was purified to homogeneity in a four-step procedure involving polyethylene glycol precipitation, hydrophobic interaction FPLC, and ion exchange FPLC. This procedure could be completed in 4 days and resulted in a greater than 900-fold purification with greater than 30% recovery. The purified enzyme exhibited a single band on a silver-stained SDS PAGE gel and had an apparent subunit molecular mass of 52 kDa. The native subunit molecular mass, determined by gel permeation FPLC, was 98 kDa, indicating that JHE from Gryllus assimilis is a dimer of two identical or similar subunits. The turnover number of the purified enzyme (1.41 s(-1)), K(M(JH-III)) (84 +/- 12 nM) of nearly-purified enzyme, and k(cat)/K(M) (1.67 x 10(7) s(-1) M(-1)) were similar to values reported for other well-established lepidopteran and dipteran JHEs. JHE from Gryllus assimilis was strongly inhibited by the JHE transition-state analogue OTFP (octylthio-1,1,1-trifluoro-2-propanone; I(50) = 10(-7) M) and by DFP (diisopropyl fluorophosphate; I(50) = 10(-7) M). The shapes of the inhibition profiles suggest the existence of multiple binding sites for these inhibitors or multiple JHEs that differ in inhibition. Isoelectric focusing separated the purified protein into 4 isoforms with pIs ranging from 4.7-4.9. N-terminal amino acid sequences (11-20 amino acids) of the isoforms differed from each other in 1-4 positions, suggesting that the isoforms are products of the same or similar genes. Homogeneously purified JHE hydrolyzed alpha-napthyl esters, did not exhibit any detectable acetylcholinesterase, acid phosphatase, or aminopeptidase activity, and exhibited only very weak alkaline phosphatase activity. JHE exhibited a low (11 microM) K(M) for long-chain alpha-naphthyl esters, indicating that JHE may have physiological roles other than the hydrolysis of JH-III. Purification of JHE represents a key step in our attempts to identify the molecular causes of genetically-based variation in JHE activity in G. assimilis. This represents the first homogeneous purification of JHE from a hemimetabolous insect.
Acetone↗
Juvenile hormone catabolism and oviposition in the codling moth, Cydia pomonella, as functions of age, mating status, and hormone treatment.
In vitro catabolism of juvenile hormone (JH) in haemolymph of adult female Cydia pomonella was ascribed mainly to juvenile hormone esterase (JHE) activity. No significant differences were noted between virgin and mated females 0-96 h post-emergence. Changes in JHE activity did not appear dependent upon fluctuations in JH titre; conversely, changes in JHE activity could not explain the changes in JH titres. Maximal JHE activity was recorded at 24 h (331.47 +/- 47.25 pmol/h/microl; 355.93 +/- 36.68 pmol/h/microl, virgin; mated insects, respectively) and preceded the peak in JH titres at 48 h. Topical application of JH II (10 ng-10 microg) or fenoxycarb (50 ng) enhanced JHE activity up to 640 and 56%, respectively. Treatment upon emergence with 10 microg JH II induced enzymic activity for less than 24 h, and when 10 microg JH II or 50 ng fenoxycarb were applied, circulating JH titres returned to control levels within 24 h. Oviposition was highly sensitive to exogenous JH and declined significantly with dosages >100 pg. To allow a degree of oocyte maturation before JH treatment, the hormone was administered at 6, 12, 24, or 48 h post-emergence and/or females were mated. Neither measure "protected" the system; oviposition declined immediately after JH application.
Acetone↗
Aspirin survival in human blood modulated by the concentration of erythrocytes.
In vitro aspirin hydrolysis rates were measured in fresh human whole blood and in its separate components. The half-life of aspirin in whole blood was relatively rapid (mean 22.2 +/- 3.9 minutes) and exhibited a significant negative correlation with hematocrit (r = -0.96). Hydrolysis of aspirin in buffer that contained only washed red blood cells (40%) was significantly more rapid (mean half-life 17.5 +/- 2.0 minutes) than that in whole blood. When aspirin was incubated in solutions of washed red blood cells that contained human serum albumin in various concentrations, the aspirin half-life was found to vary directly with the concentration of albumin used; at normal levels of albumin, the hydrolysis rate of aspirin approximated that measured in whole blood. The presence of diisopropyl fluorophosphate in low concentrations (0.02-0.05 mM) markedly inhibited the rate of aspirin hydrolysis in washed red blood cells and whole blood. We conclude that enzymes(s) linked to the erythrocyte, probably membrane-bound acetylcholinesterase, control the survival of aspirin in blood.
Aspirin↗
Rheumatoid synovial fibroblast adhesion to human articular cartilage. Enhancement by neutrophil proteases.
OBJECTIVE: To determine if preexposure of human articular cartilage to activated neutrophils alters rheumatoid synovial fibroblast adhesion to human articular cartilage. METHODS: Human articular cartilage was exposed to either activated neutrophils, interleukin-1, or supernatants obtained from activated neutrophils that had been treated with different protease inhibitors. Radiolabeled rheumatoid synovial fibroblasts were then incubated with the cartilage and the number of counts associated with the cartilage was determined. RESULTS: Pretreatment of human articular cartilage with either activated neutrophils or supernatants obtained from activated neutrophils enhanced subsequent rheumatoid synovial fibroblast adhesion. In contrast, interleukin-1 treatment of cartilage did not alter the adhesion of synovial fibroblasts. The enhanced adhesion could be attenuated by pretreatment of the neutrophil supernatants with either diisopropylfluorophosphonate or EGTA and almost completely abolished by using both inhibitors. CONCLUSION: This study demonstrates that adhesion of rheumatoid synovial fibroblasts to human articular cartilage can be enhanced by exposing the cartilage to proteases released by neutrophils. These results suggest that neutrophil products may play a role in enhancing adhesion of rheumatoid synovium to cartilage in vivo.
Adult↗
Activation of carboxylesterases in root cortical parenchymal cells of Pisum sativum during xylem induction in vitro.
A quantitative cytochemical study of naphthol AS-D esterase activity in explants from roots of Pisum sativum grown on basal medium for 3 or 6 days showed similar levels of activity to those seen in sections of cortex and stele from intact roots of similar ages. Explants grown in the presence of auxins or cytokinin alone showed a threefold or twofold increase in cortical parenchymal activity, respectively. On adding both hormones to initiate xylem element formation, there was also a threefold increase in activity in the cortex. In all three cases, the stimulated activity was totally inhibited by either 10(-4) M diisopropylfluorophosphate (DFP) or 10(-4) M diethyl p-nitrophenylphosphate (E600), indicating carboxylesterase activity. The low level of activity normally present in cortical cells was inhibitor resistant, so indicating acetylesterase activity. Thus, auxins and cytokinins appear to activate mainly similar carboxylesterases during the initiation of xylem elements from cortical parenchyma cells.
2,4-Dichlorophenoxyacetic Acid↗
Comparison of the localization of acetylcholinesterase and non-specific cholinesterase activities in mammalian and avian retinas.
Explore the source record for details and available documents.
Acetylcholinesterase↗
Acetylcholinesterase staining in subdivisions of the cat's inferior olive.
The present paper describes a unique distribution of true AChE activity in the IO. In the dorsal accessory olive three areas with high AChE activity can be distinguished. The medial accessory olive can be subdivided into a caudal part which shows rostro-caudally directed bands with different enzymatic activity, and a rostral part which shows a more uniform, medium activity. In the nucleus beta and the dorso-medial cell column, AChE activity is low. The ventral and dorsal lamellae of the principal olive contain areas with high, medium, and low activity. The dorsal cap is strongly positive, while the ventrolateral outgrowth is negative for AChE. Enzyme distribution cannot be fully explained on the base of the known afferent and efferent connections with the IO. However, histochemical results provide evidence that generally supports a subdivision of the IO that mirrors these connections (Brodal, '40; Armstrong et al., '74' Boesten and Voogd, '75; Groenewegen et al., '75).
Acetylcholinesterase↗
Identification of acetylcholinesterase-reactive neurons and neuropil in neostriatal transplants.
To identify and describe neurons in neostriatal transplants that synthesize acetylcholinesterase (AChE), the present study has utilized the irreversible AChE inhibitor diisopropylfluorophosphate (DFP) combined with AChE histochemistry. Dissociated suspensions of tissue taken from the striatal ridge of embryos at 14 days of gestation were transplanted into the neostriatum of adult rats 5 days after intrastriatal kainic acid lesions. Two types of AChE neurons have been identified in transplants treated with DFP. One type resembled the large intensely reactive AChE neuron that is thought to be a cholinergic interneuron of the normal neostriatum. The other type resembled smaller, less reactive AChE neurons of the neostriatum, as well as medium-sized, lightly reactive AChE neurons of the globus pallidus. Qualitative observations suggest that these less reactive AChE neurons were more numerous in transplants compared to the normal neostriatum. Both AChE neuronal types were found in segregated clusters throughout the grafts. Transplants processed for AChE histochemistry without DFP treatment contained two types of AChE neuropil. Dark areas of AChE neuropil similar in intensity to the normal neostriatum were found between larger areas of lighter AChE neuropil. These results demonstrate that neostriatal transplants contain AChE neurons and suggest that these neurons contribute to the AChE reactivity within the graft.
Acetylcholinesterase↗
Inability to block the activity of guinea pig migration inhibitory factor with di-isopropyl phosphorofluoridate.
Explore the source record for details and available documents.
Animals↗
A specific serine proteinase is inducible in Lyt-2+,L3T4- and Lyt-2-,L3T4+ T cells in vitro but is mainly associated with Lyt-2+,L3T4- effector cells in vivo.
Recently, we and others reported on the expression of a serine proteinase in long-term cultured murine T lymphocyte cell lines. In an attempt to explore the distribution and possible regulation of this enzyme in T lymphocyte subsets, we performed the presented detailed study. We found that the proteinase is not expressed by thymocytes and resting T cells but can be induced by lectin or antigen in combination with lymphokine sources in vitro in macrophage-depleted unselected T cells as well as in both T cell subsets (Lyt-2+,L3T4- and Lyt-2-,L3T4+) separated by flow cytofluorometry. Furthermore, it appears that cell-associated proteinase activity is increasing with prolonged culture period of sensitized T lymphocytes and that it is higher in antigen-activated as compared to lectin-activated T cells. When tested for substrate specificity the T cell-associated proteinase was shown to preferentially cleave model peptide substrates carrying L-arginine at position P1 in combination with nonpolar amino acids at position P2 and P3. As concluded from its sensitivity to proteinase inhibitors the enzyme can be classified as a serine proteinase and by molecular sieving at high ionic strength it was shown to have a mol. mass of approximately 50-60 kDa. Analysis of in vivo activated T cells revealed that this particular proteinase was expressed in flow cytofluorometry sorted lymphocytic choriomeningitis virus-specific Lyt-2+,L3T4- cytolytic T lymphocytes but not in Lyt-2-,L3T4+ T cells presensitized with either Listeria monocytogenes or I-A alloantigens. The data demonstrate that the two T cell subsets (Lyt-2+,L3T4-; Lyt-2-,L3T4+) have distinct in vitro induction requirements for the expression of proteinase and that after activation of T cells in vivo the enzyme is preferentially associated with Lyt-2+,L3T4- effector cells.
Animals↗
The T cell-specific serine proteinase TSP-1 is associated with cytoplasmic granules of cytolytic T lymphocytes.
This study describes the localization of the previously purified T cell-specific serine proteinase, termed TSP-1 (M. M. Simon et al., EMBO J. 1986. 5: 3267), within cytoplasmic granules of cytolytic T cell lines (CTLL). Subcellular fractionation of disintegrated CTLL (ruptured by nitrogen cavitation) was accomplished by Percoll density gradient centrifugation of cell lysates (postnuclear supernatant). Individual fractions were tested for proteinase activity on chromogenic peptide substrates and for the presence of TSP-1 by Western blot analysis. In addition, each fraction was assayed for cytolytic activity against sheep red blood cells (SRBC), for protein and for additional marker enzymes to assess the enrichment for cellular organells. All serine enzyme-type molecules including TSP-1 expressed by CTLL were identified by labeling cell lysates or gradient fractions with the serine proteinase-specific affinity ligand tritiated diisopropyl fluorophosphate [( 3H]DFP) in the presence or in the absence of class-specific or enzyme-specific proteinase inhibitors and subsequent sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The data demonstrate that the Percoll gradient fraction, which was shown by morphological examination in the electron microscope to be highly enriched for cytoplasmic granules, also contained greater than 80% of proteinase activity in addition to the granule-associated structures cytolysin and arylsulfatase. The identity of the granule-associated proteinase in two independent cell lines, CTLL HY3-Ag3 and CTLL 1.D.9, with the serine proteinase TSP-1 is indicated by its specificity for the chromogenic substrate H-D-Pro-Phe-Arg-p-nitroanilide, its sensitivity to class-specific as well as TSP-1-specific enzyme inhibitors and by its reactivity with a polyvalent TSP-1-specific rabbit antiserum. Both CTLL contain a [3H]DFP-labeled protein that migrates with a molecular mass of 60 kDa under nonreducing conditions and with 30 kDa under reducing conditions and which can be inactivated by the TSP-1-specific inhibitor H-D-Pro-Phe-Arg-chloromethylketone. CTLL HY3-Ag3 (a long-term culture CTLL with natural killer-like activity) but not CTLL 1.D.9 (an antigen-specific short-term cultured CTLL) express in addition a further [3H]DFP-binding protein which migrates with 27 kDa under nonreducing or reducing conditions. No substrate specificity was found for this molecule. The possible function of the granule-associated serine proteinase TSP-1 is discussed.
Amides↗