Transient‐phase and steady‐state kinetics of lipase‐catalyzed ester hydrolysis 1

Abstract

An experimental study has been made of both the steady‐state and the transient‐phase (presteady‐state) kinetics of the hydrolyses of several saturated aliphatic esters of p‐nitrophenol catalyzed by wheat germ lipase. The analysis of the presteady‐state part revealed two transients indicating that lipase‐catalyzed reactions proceed via a two‐intermediate mechanism suggested for other esterases. The possibility of more than one species of the enzyme engaged in catalytic activity is discussed and a reaction mechanism scheme is proposed accordingly.     

The mechanism of delayed neuropathy caused by some organophosphorus esters: Using the understanding to improve safety

Abstract

The mechanism of delayed neurotoxicity of some OP (organophosphorus) esters such as DFP (di‐isopropyl phosphorofluoridate) and TOCP (tri‐o‐cresyl phosphate) involves an initial two‐step process affecting an esterase called NTE (neurotoxic esterase). This understanding permits the assessment of delayed neuropathic potential in terms of a quantitative measurement of inhibition of NTE in tissue taken from dosed hens. Structure/activity relationships have been rationalized and the neurotoxic potential of those OP esters which are direct inhibitors of esterases may now be assessed in vitro. The response of human NTE can usefully be compared with that of hen NTE. Nil delayed neurotoxic potential is associated with carbamate or phosphinate anticholinesterases which may be designed as insecticides.     

Residues of 2,4‐D in air samples from Saskatchewan: 1966–1975

Abstract

Residues of 2,4‐D (2,4‐dichlorophenoxyacetic acid) in air samples from several sampling sites in central and southern Saskatchewan during the spraying seasons in the 1966–68 and 1970–75 periods were determined by gas‐liquid Chromatographic techniques. Initially, individual esters of 2,4‐D were characterized by retention times and confirmed further by co‐injection and dual column procedures. Since 1973, however, only total 2,4‐D acid levels in air samples have been determined after esterification to the methyl ester and confirmed by gc/ms techniques whenever possible.

Up to 50% of the daily samples collected during the spraying season at any of the locations and during any given year contained 2,4‐D, with butyl esters being found most frequently. The daily 24‐hr mean atmospheric concentrations of 2,4‐D ranged from 0.01 to 1.22 μg/m³, 0.01 to 13.50 μg/m³, and 0.05 to 0.59 μg/m for the iso‐propyl, mixed butyl and iso‐octyl esters, respectively. Even when the samples were analysed for the total 2,4‐D content, i.e. from 1973 onwards, the maximum level of the total acid reached only 23.14 μg/m. In any given year and at any of the sampling sites, about 30% of the samples contained less than 0.01 μg/m³ of 2,4‐D. In another 40% of the samples, the levels of 2,4‐D ranged from 0.01 to 0.099 yg/m. Only about 30% of the samples contained 2,4‐D concentrations higher than 0.1 μg/m³, with only 10% or less exceeding 1 μg/m³.

None of the samples, obtained with the high volume particu‐late sampler, showed any detectable levels of 2,4‐D, indicating little or no transport of 2,4‐D adsorbed on dust particles or as crystals of amine salts.     

Biochemical effects of some organo‐phosphorus insecticides on new targets in white rats

Abstract

The three S‐n‐propyl phosphates and phosphothioates: RH 218, profenofos and prothiophos were compared with fenitrothion in their potential as inhibitors of rat liver and brain AChE. Fenitrothion was more potent as an inhibitor than the three S‐n‐propyl derivatives. Incubation of hepatic protein enhanced ChE inhibition in brain in the case of fenitrothion, while it reduced the inhibitory effect of the S‐n‐propyl derivatives. On the other hand, the four organophosphorus esters caused hypoglycemia in both male and female rats and also reduced their blood urea with different degrees.     

Comparative toxicity of some pesticides on human health and some aquatic species

Abstract

Inhibition studies have been carried on the hydrolysis reactions catalyzed by cholinesterases taken from electric eel, human and horse serums by Sevin, Aldrin and Malathion. Three esters of p‐nitrophenol have been used as substrates. The degree of inhibition has been used as a measure of the comparative toxicity of these pesticides on these cholinesterases.     

A new synthesis of tetrachlorofluorodibenzo‐p‐dioxin

Abstract

The 1,2,3,4‐tetrachloro‐7‐fluorodibenzo‐p‐dioxin has been synthesized via condensation of 4‐fluorocatechol and pentachloronitrobenzene. This compound could be used as an internal standard for the analysis of 2,3,7,8‐tetrachlorodibenzo‐p‐dioxin by Chromatographic methods.     

Sorption and photolysis studies in soil and sediment of the herbicide napropamide

Abstract

The influence of soil and sediment composition on sorption and photodegradation of the herbicide napropamide [N, N‐diethyl‐2‐(1‐naphthyloxy)propionamide] was investigated. Five soils and one sediment were selected for this study and the clay fractions were obtained by sedimentation. Sorption‐desorption was studied by batch equilibration technique and photolysis in a photoreactor emitting within 300–450 nm wavelenght with a maximum at 365 nm. Sorption increased with clay content and was not related to organic matter *content. High irreversibility of sorption was related to the greater montmorillonite content. The presence of soil or sediment reduced photolysis rate due to screen effect and this process did not depend on solid composition but on particle size distribution.     

Applications of isotope differentiation for metabolic studies with di‐(2‐ethylhexyl) phthalate

Abstract

The pervasiveness of the plasticizer di‐(2‐ethylhexyl) phthalate (DEHP) in the environment and especially in the laboratory results in a background that may cause severe interference with analytical studies. Animal‐to‐animal variability in the distribution of DEHP metabolites in excreta normally makes it necessary to use large groups of animals when different treatments are compared. Finally, radioactive tracers are usually considered undesirable for metabolic studies involving human subjects. All of these problems can be overcome through the use of muliple isotopic labels, especially ¹²C/¹³C/¹⁴C. Examples are given involving rats and monkeys, and applicability to humans is discussed. The principles involved are not limited to any particular class of test compounds. In rats, the competing pathways for metabolism of phthalate esters produce a different distribution of metabolites from a small intravenous dose of DEHP than from a large oral dose.     

Excretion balance,metabolic fate and tissue residues following treatment of rats with amitraz and N‘‐ (2,4‐dimethylphenyl)‐N‐methylformamidine

Abstract

Amitraz and its metabolite N‘‐ (2,4‐dimethylphenyl)‐N‐methyl‐formamidine (BTS‐27271) were administered orally to white rats. Both compounds were rapidly metabolized and eliminated primarily via the urine. The cumulative percentage of the dose eliminated in the urine was 77.6 for amitraz and 88.7 for BTS‐27271 by 96 hr posttreatment. Amitraz degradation products present in urine included BTS‐27271, 2,4‐dimethylformanilide, 2,4‐dimethylaniline, 4‐formamido‐3‐methylbenzoic acid, 4‐amino‐3‐methylbenzoic acid, and several unknowns. BTS‐27271 degradation products in rat urine were similar to those found with amitraz. Tissue residues generally were low (<25 ppb) with the exception of those in liver.     

Fate and residues of 14C‐chloramphenicol in laying chickens 1

Abstract

Studies were conducted to determine the metabolic fate of chloramphenicol (CAP) in White Leghorn using the ^l4C‐labelled compound. In one experiment birds were administered orally via intra‐crop, a single dose of 100 mg (equivalent to 66 mg kg^‐1 body weight) of CAP containing 14 μCi ¹⁴C‐CAP, and its absorption, elimination and distribution in plasma were recorded. Orally dosed ¹⁴C‐compound was rapidly absorbed, efficiently distributed in plasma and eliminated in excreta (>70% in 5 hr). After 5 h, CAP equivalent residues in tissues were lower than 15 μg g^‐1 for this treatment. In a second experiment birds were given intra‐crop dose of either 0.5 or 5 mg of CAP (each dose contained 2.5 μCi ¹⁴C‐CAP) daily for five consecutive days followed by a seven day withdrawal period and elimination of ¹⁴C in excreta and eggs was monitored. More than 95% of the administered ¹⁴C was eliminated within the first 24 h after dosing. Radiocarbon (¹⁴C) was deposited preferentially in yolks compared to albumen or other tissues. Residues declined when feeding was stopped. Various metabolites were isolated and identified by a combination of TLC, LC, and LC‐MS. The main metabolic route of CAP in laying hens appears to be the glucuronidation. Cleavage of the dichloroacetate moiety was only a minor route.     

Aerobic degradation of diuron by aquatic microorganisms

Abstract

Degradation of diuron [3‐(3,4‐dichlorophenyl)‐l,1‐dimethyl‐urea] by microorganisms obtained from pond water and sediment was determined under aerobic conditions. Enrichment procedures were used to isolate cultures capable of degrading the herbicide. Several mixed fungal/bacterial and mixed bacterial cultures were isolated that could degrade diuron. The mixed cultures degraded 67–99% of the added diuron forming from six to seven products which were separated via TLC. The major degradation product detected in most culture extracts was 3,4‐dichloroaniline. Other identified products formed were 3‐(3,4‐dichlorophenyl)‐1‐methylurea and 3‐(3,4‐dichlorophenyl)urea.     

Leaching,mobility and persistence of tebufenozide in columns packed with forest litter and soil

Abstract

Leaching, downward mobility and persistence of tebufenozide was investigated under laboratory conditions in columns packed with forest litter and soil, after fortification with the analytical grade material (purity > 99.6%) and with two commercial formulations, RH‐5992 2F (aqueous flowable) and RH‐5992 ES (emulsion suspension). Two types of litter and soil were used: one type with relatively high amounts of sand and the other with high amounts of clay.

The concentrations eluted in the leachates were lower when the analytical material (dissolved in acetone) was used for fortification, than when the two formulations (diluted with water) were used. The amount leached was higher for RH‐5992 2F than for RH‐5992 ES. The type of substrate, i.e., sandy or clay type, had only marginal influence on the amounts eluted in the leachates. Downward movement of tebufenozide from the top 2‐cm layer to the untreated middle and bottom layers (3‐cm segments) was consistently lower when the analytical material was used for fortification, than when the two formulations were used. Downward movement was higher for RH‐5992 2F than for RH‐5992 ES. Persistence of tebufenozide in substrates, maintained under submerged conditions for 70 days after leaching, indicated an initial 2‐week lag period prior to the onset of degradation. Formulation‐related differences were observed in the half‐life (DT₅₀) values. When the analytical material was used for fortification, the DT₅₀ ranged from ca 54 to 59 d. However, when the formulations were used for fortification, the DT₅₀ showed a higher range, i.e., from ca 62 to 67 d for RH‐5992 2F and ca 70 to 80 d for RH‐5992 ES. Formulation ingredients appear to have caused enhanced adsorption of tebufenozide onto the substrates, thus delaying degradation.     

Comparative study of the linearity and sensitivity of electron capture and flame photometric detectors using a pesticide standard

Abstract

A study was undertaken to compare and evaluate the different types of ⁶³Ni and ³H electron capture (ECD) and flame photometric detectors (FPD) as part of the Canadian Check Sample Program on Pesticide Residue Analysis. Twenty‐seven laboratories were supplied with chlorpyrifos standard solutions for the determination of linear range and minimum detectable amounts (MDA). Each laboratory selected its own operating conditions except for the FPD study. Results for the FPD were compared with standardized flame conditions designed to optimize the exponential factor in the S‐mode. The MDA of the ECDs (pulsed and direct‐current) ranged from 0.09 to 0.17 pg/sec for ⁶³Ni and ³H sources respectively. The corresponding MDA for the linearized ECDs (pulse modulated or constant current) was 0.04 pg/sec and a linear range of 10⁴ or greater with nitrogen as the carried gas. Use of argon/methane and a pulse width of 0.1 usec extended this range to 10⁵ but the sensitivity was not as great as with nitrogen carrier gas. The practical limit of detector sensitivity was found to be instrument dependent depending primarily on the noise level. In the FPD study, operating in the P‐mode gave an average MDA of 5.97 pg/sec and a linear range of 2 x 10³. Similarly, the S‐mode parameters were 81.1 pg/sec and 8 x 10² respectively with an average exponential factor of n 1.8. Attempts to apply optimized detector gas‐flow conditions improved sensitivity and linearity in the S‐mode only.     

Chemical evaluation of SZI‐121, a tetrazine type miticide

Abstract

SZI‐121, a tetrazine type acaricide, has been improved by Chinoin. The compound is very active against phytophagous mites and has good toxicological and environmental properties. The chemical structure was designed to get improvements in mode of action and efficacy simultaneously. SZI‐121 acts not only as a contact ovicide but due to very good translaminar activity it can enter mites via ingestion. SZI‐121 possesses transovarian activity and it also able to stop the development of mites at chrysalis stages.     

A study on the effect of deoxynivalenol on serotonin receptor binding in fig brain membranes

Abstract

Central serotoninergic (5‐hydroxytryptamine, 5HT) pathways are believed to be involved in the mechanisms of anorexia and/or emesis evoked by the trichothecene mycotoxin deoxynivalenol (DON). Using an in vitro membrane receptor binding assay, the competitive potency of DON was investigated against several radioactive ligands that have a high affinity for selective 5HT‐receptor subgroups. Receptor site densities and displacement profiles in twelve selected regions of pig brain were investigated. Overall, DON possessed only minimal efficacy to competently block any of the 5HT‐ligands tested. IC₅₀ values (50% inhibitory concentration) of at least 5 mM DON were required to inhibit binding, and in certain regions concentrations of 100 mM were ineffective. In comparison, several standard 5HT‐antagonists showed 10³‐10⁵ times greater capability than DON to displace binding of these ligands. Because these results indicated DON possesses only weak affinity for the 5HT‐receptor subtypes investigated here, this suggested that in vivo, unless relatively high concentrations of the toxin are present, its pharmacological effects may be mediated by mechanisms other than a functional interaction with serotoninergic receptors at the central level.     

Lung subcellular fractions and surfactant lipid metabolism of rats exposed with DDT or endosulfan intratracheally

Abstract

The effect of intracheally administered DDT (5 mg/100 g body weight) or endosulfan (1 mg/100 g body weight) for three cosecutive days have been studied on lipid metabolism of rat lung subcellular fractions. Both the insecticides did not affect the lung weight and the protein contents of microsomes, lamellar bodies and surfactant but significantly increased the phospholipid contents of microsomal and surfactant system. Most of the neutral lipid components of lung subcellular fractions were also increased by DDT or endosulfan treatments, except that of surfactant triglycerides which were decreased by DDT treatment. DDT or endosulfan both increased the incorporation of radioactive [methyl‐³H]choline into microsomal phosphatidylcholine (PC) and surfactant dipalmitoylphosphatidylcholine (DPPC) without affecting the incorporation of radioactive [methyl‐¹⁴C]methionine, showing the increased synthesis of PC via CDPcholine pathway. The results presented in this communication showed that DDT and endosulfan, the two different chloroinsecticides have similar effects on microsomal lipid metabolism but produce different biochemical manifestations on the secretion of surfactant phospholipids.     

Toxic effects of carbofuran and glyphosate on semen characteristics in rabbits

Abstract

The present study was undertaken to investigate the effect of chronic treatment with two sublethal doses of Carbofuran (carbamate insecticide) and Glyphosate (organophosphorus herbicide) on body weight and semen characteristics in mature male New Zealand white rabbits. Pesticide treatment resulted in a decline in body weight, libido, ejaculate volume, sperm concentration, semen initial fructose and semen osmolality. This was accompanied with increases in the abnormal and dead sperm and semen methylene blue reduction time. The hazardous effect of these pesticides on semen quality continued during the recovery period, and was dose‐dependent. These effects on sperm quality may be due to the direct cytotoxic effects of these pesticides on spermatogenesis and/or indirectly via hypothalami‐pituitary‐testis axis which control the reproductive efficiency.     

Herbicide safeners: Tools for improving the efficacy and selectivity of herbicides

Abstract

Safeners (also known as antidotes) are chemical or biological agents that reduce the toxicity of herbicides to crop plants by a physiological or molecular mechanism. Commercialized safeners are mainly chemical compounds that enhance the tolerance of selected grass crops such as maize (Zea mays L.), grain sorghum [Sorghum bicolor (L.) Moench], rice (Oryza sativa L.), and wheat (Triticum aestivum L.) to chloroacetanilide, thiocarbamate, sulfonylurea, imidazolinone, and aryloxyphenoxypropionate herbicides. In practice, safeners are applied either to the crop prior to planting (seed safeners) or to the soil together with the herbicide, formulated as a prepackaged mixture. Safeners act as "bioregulators”; controlling the amount of a given herbicide that reaches its target site in an active form. A safener‐induced enhancement of the metabolic detoxification of herbicides in protected plants is the most apparent mechanism for the action of all commercialized safeners. Herbicide‐detoxifying enzymes such as glutathione transferases (GST), cytochrome P‐450 monooxygenases (CytP450), esterases, and UDP‐glucosyltransferases are induced by herbicide safeners. At the molecular level, safeners appear to act by activating or amplifying genes coding for these enzymes (e.g., GST).     

Cobalt induced changes in immune response and adenosine triphosphatase activities in rats

Abstract

The immuno‐biochemical effects of cobaltous chloride in rats receiving iron‐sufficient and deficient diets were investigated. Rats receiving 100 ppm or more cobalt showed a significant reduction in thymus and body weights along with a marked decrease in hemoglobin, hematocrit, sheep agglutinins and plaque forming cells. These effects were more pronounced in rats receiving cobalt mixed with iron‐deficient diet than those fed on iron‐sufficient diet. The Na⁺‐K⁺ and mitochondrial (Oligomycin‐sensitive) Mg²+ATPase activities in brain and liver of rats fed with iron‐deficient diets were decreased significantly. However, the ATPase activities in these tissues from rats fed with cobalt mixed with iron‐sufficient diets were not altered.     

Changes in lipid profiles of liver microsomes of rats following intratracheal administration of DDT or endosulfan

Abstract

The effect of intratracheal administration of DDT (5 mg/100 g body weight) or endosulfan (1 mg/100 g body weight) to rats for three consecutive days, has been studied on liver lipid metabolism. The administration of DDT but not endosulfan significantly increased the liver weight and the microsomal protein contents. Both DDT and endosulfan treatments significantly increased the contents of microsomal phosphatidylcholine (PC), total‐free‐ and esterified cholesterol. The distribution of unsaturated fatty acids of microsomal PC and PE was increased by DDT treatment. The intratracheal administration of DDT caused fatty infiltration of liver which was probably due to increased synthesis of triglycerides (TG). This is supported by the increased incorporation of radioactive palmitate‐l‐¹⁴C into microsomal TG. However, the increased incorporation of palmitate‐l‐¹⁴C into microsomal PC and phosphatidylethanolamine (PE) after the DDT treatment, was due to the increased transacylation reaction supported by the decreased activity of microsomal phospholipase A. The intratracheal adminstration of endosulfan did not has pronounced effect on liver fatty infiltration, or transacylation reaction in microsomal PC and PE. However, the results have shown that the treatments of DDT or endosulfan increased the PC contents and the incorporation of radioactive [methyl‐³H]choline into PC of microsomes, resulting the increased synthesis of PC via CDPcholine pathway. Thus, the intratracheally administered DDT or endosulfan to rats showed that both the insecticides cause manifestations in the biochemistry of microsomal membrane lipids, although the effects of DDT being more pronounced. Therefore, the translocation effects of these insecticides or metabolites from lung to liver is established.