Biochemical alterations in skeletal muscle induced by 2,4-dichlorophenoxyacetic butyl ester during chick embryonic development

2,4-Dichlorophenoxyacetic butyl ester (2,4-D b.e.) (3.1 mg/egg) was applied on fertile hen eggs before starting the incubation. Chicks hatched from treated eggs showed motor dysfunctions, postural troubles and edematous muscles. The electromyography revealed muscular weakness, prolonged motor distal latency, and myotonia. The biochemical composition of leg and complexus muscles from 1-day-old chicks was determined. A significant diminution (24%) in the glycogen level of leg muscles was produced by the treatment. There was a small increase (15%) in sarcoplasmic proteins from leg muscles and an increase of a 20 kD protein in the myofibrillar proteins from complexus muscles. Even though total lipid content was not changed, 2,4-D b.e. treatment produced a diminution of sterol esters (20%) and phosphatidylcholine (11%) and an increase of phosphatidylserine (61%), triglycerides (37%) and free fatty acids (FFA) (448%) in leg muscles. Increases of phosphatidylethanolamine (16%), sterols (58%) and FFA (267%) were detected in complexus muscles. A remarkable increase (700-1500%) of unsaturated FFA, e.g. oleic, linoleic and arachidonic acids, was observed. Considering the avian embryo lipid metabolism, it is proposed that FFA and triglycerides were accumulated because they could not be metabolized in the mitochondria. Since FFA are potent cytotoxic compounds, their increase may be a key factor in the 2,4-D b.e. toxic action in muscle and other tissues during embryonic development.     

赛庚啶对犬心肌线粒体膜ATP酶活性和~(45)Ca~(2+)摄取功能的影响

结果表明,本实验条件下制备的心肌线粒体基本上没有细胞膜及肌浆网的污染 赛庚啶(CyP)对正常犬心肌线粒体膜ATP酶活性有一定的抑制作用,其IC_(50)为O.839mmol·L~(-1).正常犬心肌线粒体的~(45)Ca~(2+)摄取至第10min趋于稳定,其Ca~(2+)摄取量可达141±s 22 nmol·mg~(-1)蛋自;Cyp对心肌线粒体的~(45)Ca~(2+)摄取过程有较强的抑制作用,其IC_(50)为31μmol·L~(-1)。     

Effects of 2,4-dichlorophenoxyacetic acid on Rhizobium sp. in pure culture

The effects of 2,4-Dichlorophenoxyacetic acid on the growth rate, chemical composition, ¹⁴C-2,4-dichlorophenoxyacetic acid and ⁴⁵Ca²⁺ uptake by Rhizobium sp. M 4 able to nodulate Arachis hypogaea were determined. Cellular growth was diminished by the presence of 10^?3 M 2,4-dichlorophenoxyacetic acid in the medium. Alterations in cellular chemical composition, in ¹⁴C-2,4-dichlorophenoxyacetic acid and in ⁴⁵Ca²⁺ uptake were found.     

Nucleic acid content and residue determination in tissues of chicks born from 2,4-dichlorophenoxyacetic butyl ester treated eggs

The effects of 2,4-dichlorophenoxyacetic butyl ester (2,4-D b.e.) on protein and nucleic acid contents of different tissues of chicks born from externally treated hen's eggs were studied. Residues of 2,4-D b.e. were also determined and quantified by gas liquid chromatography (GLC). A diminished DNA content was observed in skeletal muscle and stomach, an increased one was found in cerebrum and cerebellum, whereas no variations were observed in the other analysed tissues. Protein levels were found decreased in kidney, cerebrum and heart. Besides, the GLC determinations revealed the presence of the 2,4-D b.e. in all the studied tissues finding the highest concentrations in kidney. 2,4-D did not appear to be a significant metabolite of 2,4-D b.e. in this experimental system. The dissimilar effects of 2,4-D b.e. on protein and nucleic acid contents of the studied tissues would be related to a different tissue susceptibility to the drug and/or to drug distribution.     

Dissimilation of 2,4-dichlorophenoxyacetic acid by Azotobacter chroococcum

1. A strain of Azotobacter chroococcum which could use 2,4-dichlorophenoxyacetic acid (2,4-D) as sole carbon source was isolated. 2. The strain metabolized 2,4-D via p-chlorophenoxyacetic acid, p-chlorophenol and 4-chlorocatechol; the last metabolite was cleaved by catechol 1,2-dioxygenase. 3. The enzyme exhibited broad substrate specificity.     

Stimulation of canine cardiac sarcoplasmic reticulum Ca2+ uptake by dihydropyridine Ca2+ antagonists

We examined the effects of four Ca2+ antagonists that possess the ability to bind to calmodulin-felodipine, nitrendipine, prenylamine, and verapamil--as well as the effect of the calmodulin antagonist trifluoperazine on Ca2+ uptake and Ca2+ + Mg2+/ATPase activity in canine cardiac sarcoplasmic reticulum. In the presence of 20-30 microM felodipine and 100-200 microM nitrendipine, Ca2+ uptake increased from 69 nmoles X mg-1 X min-1 to 107 and 108 nmoles X mg-1 X min-1, respectively, with half-maximal stimulation occurring at 7.5 and 28 microM respectively. Ca2+ + Mg2+/ATPase activity was unchanged over the same concentration ranges. In contrast, both Ca2+ uptake and Ca2+ + Mg2+/ATPase activities were inhibited in the presence of 10-100 microM trifluoperazine (IC50 = 25 microM), 10-100 microM prenylamine (IC50 = 35 microM) and 100-200 microM verapamil (inhibition insufficient for IC50 determination). None of the drugs affected membrane permeability to Ca2+ as determined by passive 45Ca2+ efflux in the presence of ethyleneglycol bis(beta-amenoethyl ether)N,N,N1-tetraacetic acid (EGTA). Drug inhibition of calmodulin-dependent turkey gizzard myosin light chain kinase activation in a purified protein system was used as a direct measure of calmodulin antagonism, and felodipine, nitrendipine, trifluoperazine, prenylamine, and verapamil blocked this activation at IC50 values of 9.8, 55, 6.4, 31, and 93 microM respectively. None of the drugs studied, however, had any effect upon endogenous phospholamban phosphorylation in our cardiac sarcoplasmic reticulum preparations. These observations indicate that dihydropyridine Ca2+ antagonists stimulate cardiac sarcoplasmic reticulum Ca2+ uptake in vitro either by increasing the efficiency of the transport process or by inhibiting Ca2+-dependent Ca2+ release, and suggest that these effects do not result from interference with calmodulin-mediated processes.     

Radioimmunoassay for 2,4-dichlorophenoxyacetic acid

Antisera to 2,4-dichlorophenoxyacetic acid (2,4-D), a widely used herbicide, have been obtained from rabbits following immunization with various 2,4-D-protein conjugates. Employing [¹²⁵I] 2,4-D-tyramine as the radioligand for the antisera, very poor assay sensitivity was achieved because of a much higher affinity of the antibodies to the tracer. When using [6-³H] 2,4-D(specific radioactivity 465 GBq/mmol) a sensitive and specific radioimmunoassay (RIA) for 2,4-D could be developed, which allows determination directly in water, plasma and urine samples. Levels as low as approximately 100 pg (450 femtomoles) of 2,4-D can be detected. The antiserum is fairly specific for 2,4-D. Other related phenoxycarboxylic acids and dichlorophenol showed a cross-reactivity smaller than 10%. After a single administration of 2,4-D (0.91 mg/100 g body weight, orally) to rats, plasma and urine levels were determined at different times. Results correspond to those found in the literature, thus indicating the utility of the RIA. Further applications and limitations are discussed.     

Transient effects of 2,4-dichlorophenoxyacetic acid (2,4-D) exposure on some metabolic and free radical processes in goldfish white muscle

This study aims to assess effects of 96 h goldfish exposure to 1, 10 and 100 mg/L of the herbicide, 2,4-dichlorophenoxyacetic acid (2,4-D), on metabolic indices and free radical process markers in white muscle of a commercial fish, the goldfish Carassius auratus L. Most oxidative stress markers and antioxidant enzymes were not affected at 2,4-D fish treatment. 2,4-D fish exposure induced the elevated levels of total (by 46% and 40%) and reduced (by 77% and 73%) glutathione in muscles of goldfish of 10 mg/L 2,4-D and recovery (after 100 mg/L of 2,4-D exposure) groups, respectively. However, in muscles of 100 mg/L 2,4-D exposed goldfish these parameters were depleted (by 47% and 64%). None of investigated parameters of protein and carbohydrate metabolisms changed in white muscles of 2,4-D exposed fish, with exception of lactate dehydrogenase activity, which was slightly (by 11–15%) elevated in muscles of goldfish exposed to 10–100 mg/L of 2,4-D, but also recovered. Thus, the short term exposure of goldfish to the selected concentrations of 2,4-D does not substantially affect their white muscle, suggesting the absence of any effect under the environmentally relevant concentrations.     

Effect of 2,4-dichlorophenoxyacetic acid on rat maternal behavior

Exposure to 2,4-dichlorophenoxyacetic acid (2,4-D) has several deleterious effects on the nervous system such as alterations in the concentrations of neurotransmitters in the brain and/or behavioral changes, myelination rate, ganglioside pattern [Bortolozzi, A., Duffard, R., Antonelli, M., Evangelista de Duffard, A.M., 2002. Increased sensitivity in dopamine D(2)-like brain receptors from 2,4-dichlorophenoxyacetic acid (2,4-D)-exposed and amphetamine-challenged rats. Ann. N.Y. Acad. Sci. 965, 314-323; Duffard, R., García, G., Rosso, S., Bortolozzi, A., Madariaga, M., DiPaolo, O., Evangelista de Duffard, A.M., 1996. Central nervous system myelin deficit in rats exposed to 2,4-dichlorophenoxyacetic acid throughout lactation. Neurotoxicol. Teratol. 18, 691-696; Evangelista de Duffard, A.M., Orta, C., Duffard, R., 1990. Behavioral changes in rats fed a diet containing 2,4-dichlorophenoxyacetic butyl ester. Neurotoxicology 11, 563-572; Evangelista de Duffard, A.M., Bortolozzi, A., Duffard, R.O., 1995. Altered behavioral responses in 2,4-dichlorophenoxyacetic acid treated and amphetamine challenged rats. Neurotoxicology 16, 479-488; Munro, I.C., Carlo, G.L., Orr, J.C., Sund, K., Wilson, R.M. Kennepohl, E. Lynch, B., Jablinske, M., Lee, N., 1992. A comprehensive, integrated review and evaluation of the scientific evidence relating to the safety of the herbicide 2,4-D. J. Am. Coll. Toxicol. 11, 559-664; Rosso et al., 2000], and its administration to pregnant and lactating rats adversely affects litter growth and milk quality. Since normal growth of the offspring depends on adequate maternal nursing and care, we evaluated the effect of 2,4-D on rat maternal behavior as well as the dam's monoamine levels in arcuate nucleus (AcN) and serum prolactin (PRL) levels. Wistar dams were exposed to the herbicide through the food from post partum day (PPD) 1 to PPD 7. Dams were fed either with a 2,4-D treated diet (15, 25 or 50mg 2,4-D/kg/daybw) or with a control diet. We observed that maternal nesting behavior was not modified by 2,4-D treatment. However, mother-pup interactions, specially the nursing behavior, were altered. Retrieval, crouching and licking of pups were reduced or suspended after 2,4-D treatment. We also observed an increase in the latency of retrieval and crouching in the dams treated with the herbicide. Dams showed movement along cage peripheries, food consumption during the light phase and high self-grooming. In addition of the deficits observed in maternal behavior parameters, increased catecholamine levels and a drastic decrease in indolamine levels in the AcN of treated dams were determined. Serum PRL levels were also diminished by 62%, 68% and 70% with respect to control dams in the 15, 25 and 50mg 2,4-D/kgbw treated dams, respectively. In conclusion, exposure to 2,4-D during the first post partum days produced changes in maternal behavior, serum prolactin and monoamine levels in the AcN of treated dams.     

Characterization of 2,4-dichlorophenoxyacetic acid transport and its relationship with polyamines in Azospirillum brasilense

We have previously shown that 2,4-dichlorophenoxyacetic acid (2,4-D) inhibits Azospirillum brasilense growth, the synthesis of DNA, RNA and proteins. These toxic effects are prevented when polyamines are added to the culture medium. The purposes of our research were to determine the effects of the herbicide on the number of viable Azospirillum brasilense cells, characterize the 2,4-D transport system and to study the effects of polyamines upon the latter in this microorganism. We found that 2,4-D reduced the number of viable cells and that 2,4-D transport is energyindependent, since it was not affected by metabolic inhibitors. Polyamines did not alter 2,4-D uptake, further supporting the hypothesis that the herbicide most likely produces its toxic effects by interfering with the polyamine metabolism.     

Review of 2,4-dichlorophenoxyacetic acid (2,4-D) biomonitoring and epidemiology

A qualitative review of the epidemiological literature on the herbicide 2,4-dichlorophenoxyacetic acid (2,4-D) and health after 2001 is presented. In order to compare the exposure of the general population, bystanders and occupational groups, their urinary levels were also reviewed. In the general population, 2,4-D exposure is at or near the level of detection (LOD). Among individuals with indirect exposure, i.e. bystanders, the urinary 2,4-D levels were also very low except in individuals with opportunity for direct contact with the herbicide. Occupational exposure, where exposure was highest, was positively correlated with behaviors related to the mixing, loading and applying process and use of personal protection. Information from biomonitoring studies increases our understanding of the validity of the exposure estimates used in epidemiology studies. The 2,4-D epidemiology literature after 2001 is broad and includes studies of cancer, reproductive toxicity, genotoxicity, and neurotoxicity. In general, a few publications have reported statistically significant associations. However, most lack precision and the results are not replicated in other independent studies. In the context of biomonitoring, the epidemiology data give no convincing or consistent evidence for any chronic adverse effect of 2,4-D in humans.     

Review of 2,4-dichlorophenoxyacetic acid (2,4-D) biomonitoring and epidemiology

A qualitative review of the epidemiological literature on the herbicide 2,4-dichlorophenoxyacetic acid (2,4-D) and health after 2001 is presented. In order to compare the exposure of the general population, bystanders and occupational groups, their urinary levels were also reviewed. In the general population, 2,4-D exposure is at or near the level of detection (LOD). Among individuals with indirect exposure, i.e. bystanders, the urinary 2,4-D levels were also very low except in individuals with opportunity for direct contact with the herbicide. Occupational exposure, where exposure was highest, was positively correlated with behaviors related to the mixing, loading and applying process and use of personal protection. Information from biomonitoring studies increases our understanding of the validity of the exposure estimates used in epidemiology studies. The 2,4-D epidemiology literature after 2001 is broad and includes studies of cancer, reproductive toxicity, genotoxicity, and neurotoxicity. In general, a few publications have reported statistically significant associations. However, most lack precision and the results are not replicated in other independent studies. In the context of biomonitoring, the epidemiology data give no convincing or consistent evidence for any chronic adverse effect of 2,4-D in humans.     

Studies on the mechanism of action of a new Ca-2+ antagonist, 8-(N,N-diethylamino)octyl 3,4,5-trimethoxybenzoate hydrochloride in smooth and skeletal muscles.

1. The rabbit aortic strip, guinea-pig ileum and rabbit skeletal muscle sarcoplasmic reticulum preparations were used to determine at which sites and in what manner 8-(N,N-diethylamino)-octyl 3,4,5,-trimethoxybenzoate (TMB-8) interferes with Ca2+ availability in smooth and skeletal muscles. 2. TMB-8 (50 muM) significantly inhibited equivalent responses of the rabbit aortic strip to KCl and noradrenaline. 3. TMB-8 (65 muM) produced no significant alteration in the extracellular space of the guinea-pig ileum as measured with [3H]-sorbitol. 4. The resting cellular Ca2+ influx as well as the resting 45Ca2+ efflux in the guinea-pig ileum preparation were significantly inhibited by TMB-8 (65 muM). 5. TMB-8 (5 muM and 50 muM) had no significant effect on the uptake of 45Ca2+ by the sarcoplasmic reticulum preparation of skeletal muscle; however, TMB-8 (5 muM) did significantly inhibit the caffeine (20 mM)-induced release of 45Ca2+ from this preparation. 6. It is concluded that TMB-8 reduces Ca2+ availability in smooth and skeletal muscles by stabilizing Ca2+ binding to cellular Ca2+ stores and thereby inhibits the release of this Ca2+ by contractile stimuli.     

Comparison of three tetramic acids and their ability to alter membrane function in cultured skeletal muscle cells and sarcoplasmic reticulum vesicles.

Cyclopiazonic acid is a potent inhibitor of calcium uptake and Ca(2+)-ATPase activity in sarcoplasmic and endoplasmic reticulum. In L6 muscle myoblasts, cyclopiazonic acid stimulates the uptake of tetraphenylphosphonium, a lipophilic membrane potential probe, and has antioxidant properties. The purpose of the present study was to investigate the structural requirements necessary for causing the surface charge alterations, and the antioxidant activity in L6 skeletal muscle myoblasts, and for inhibition of calcium transport by rat skeletal muscle sarcoplasmic reticulum vesicles. This was accomplished by comparing the effects of two structurally related tetramic acids, cyclopiazonic acid imine and tenuazonic acid, with cyclopiazonic acid. Cyclopiazonic acid imine inhibited oxalate-assisted 45Ca2+ uptake and ATPase activity in sarcoplasmic reticulum vesicles and stimulated tetraphenylphosphonium accumulation by L6 muscle myoblasts. However, these effects required an approximately fourfold higher concentration than that of cyclopiazonic acid. Tenuazonic acid, up to 1 mM, had no effect on oxalate-assisted 45Ca2+ uptake or Ca(2+)-ATPase activity in sarcoplasmic reticulum vesicles and did not stimulate tetraphenylphosphonium accumulation by L6 muscle myoblasts. Cyclopiazonic acid was only slightly more effective than cyclopiazonic acid imine at preventing the patulin-induced increase in thiobarbituric acid positive substance (used to estimate lipid peroxidation); tenuazonic acid was totally ineffective. Previously, it was shown that cyclopiazonic acid was twice as effective as cyclopiazonic acid imine at preventing increases in thiobarbituric acid positive substance in cultured renal cells, LLC-PK1. Thus, the indole nucleus of cyclopiazonic acid is essential for the membrane-associated biological activity; however, modification of the acetyl group reduces the potency of the activity.     

The metabolic fate of 2,4-dichlorophenoxyacetic acid-n-butyl ester in the wistar rat

The metabolic fate of 2,4-dichlorophenoxyacetic acid (2,4-D)-n-butyl ester in rats has not been extensively studied. Upon subcutaneous administration of a 100 mg/kg dose of 2,4-D butyl ester to four male Wistar rats, urine samples were analyzed by three analytical techniques for the presence of the butyl ester and metabolites. Thin layer chromatography (TLC), gas chromatography using an electron capture detector (GC-ECD), and gas chromatography-mass spectroscopy (GC-MS) were the techniques employed. 2,4-D butyl ester was rapidly hydrolyzed in the body to form 2,4-D acid. Ninety-five percent of the administered dose was excreted into the urine as the free acid within 48 h of injection, while only a small fraction (5%) was excreted over an additional 48 h. No amino acid conjugates or the parent 2,4-D butyl ester could be detected in the urine of treated rats. A minor metabolite (2% of dose) was detected by GC-MS analysis of urine samples. This compound appears to be a side chain metabolite of the 2,4-D butyl ester. Some chemical properties of the metabolite were characterized, and a 2,4-D hydroxyethyl ester structure proposed. The mechanism of this minor metabolic formation remains unknown.This research was supported in part by the Veterans Administration Research Program     

2,4-D toxicosis. I: A pilot study of 2,4-dichlorophenoxyacetic acid- and dicamba-induced myotonia in experimental dogs.

English Pointer dogs dosed po with encapsulated 2,4-dichlorophenoxyacetic acid (2,4-D) or 2-methoxy-3,6-dichlorobenzoic acid (dicamba) developed varying degrees of myotonia. Dogs given 175 or 220 mg of 2,4-D/kg body weight rapidly developed clinical and electromyographic (EMG) manifestations consistent with a diagnosis of myotonia or pseudomyotonia. Dogs given 2,4-D at 86.7, 43.7 or 8.8 mg/kg body weight developed subclinical manifestations of myotonia detectable only with an electromyograph. The administration of 2,4-D at 1.3 or 1.0 mg/kg body weight failed to produce detectable EMG changes. One dog given dicamba at 86.7 mg/kg body weight developed clinical and EMG manifestations of myotonia similar to those induced by the highest doses of 2,4-D.     

K+, MG2+-aspartate (KMg-ASP)--mediated prevention of isoproterenol(ISO)-induced metabolic changes in the myocardium.

Isoproterenol (ISO) in a single dose of 7.5 mg/kg b.w. i.v was found to produce alterations of cardiac metabolism in dogs. After 2 h, high energy phosphate stores and glycogen were reduced, whereas the levels of lactate, pyruvate, the lactate/pyruvate ratio and myofibrillar ATPase activity were elevated. Ca2+ -accumulation by sarcoplasmic reticulum (SR) and mitochondria were increased (P less than 0.01). After 24 h, a partial recovery in the parameters followed could be observed. Only myofibrillar ATPase activity and the Ca2+ -uptake by SR and mitochondria were lowered. When K,Mg-ASP was administered, i.v. concurrently with ISO, myofibrillar ATPase and Ca2+ -accumulation by SR did not differ from controls 2 h after ISO application. Also the other parameters exhibited a tendency to improve (P less than 0.01), but did not reach control levels. 24 h after ISO application we could observe a similar effect of K,Mg-ASP in the prevention of Ca2+ -overload accompanying metabolic changes.     

Percutaneous penetration of 2,4-dichlorophenoxyacetic acid and 2,4-D dimethylamine salt in human volunteers.

The percutaneous penetration of 2,4-dichlorophenoxyacetic acid (2,4-D) and 2,4-D dimethylamine salt (DMA) was evaluated separately in five male volunteers who participated in both experiments. Urine samples collected for 144 h following dermal applications of 10 mg to the dorsum of the hand (9 cm2) were analyzed for 2,4-D. Following the acid application, an average of 4.46 +/- 0.849% was recovered in the urine and a significantly lower amount of 1.76 +/- 0.568% following the DMA application. Significantly higher amounts of 2,4-D DMA (7.68 +/- 0.493 mg) were washed off the hand 6 h following application as compared with 2,4-D acid (5.35 +/- 0.384 mg). These results indicate that, in addition to the differences in physical and chemical properties of the two compounds that will affect absorption, the amount of the chemical absorbed is related inversely to the amount of washed off. Urinary excretion of 2,4-D was not complete in all volunteers 144 h following either application, but in all cases it was approaching the limit of detection. An average of 84.8 +/- 2.55% and 76.8 +/- 8.05% of the total recovered in 144 h was recovered in the urine 96 h following 2,4-D acid and 2,4-D DMA application, respectively. Average, approximated half-lives for excretion were 39.5 +/- 8.1 h for the acid application and 58.5 +/- 13.2 h for the DMA application.     

Dantrolene and calcium uptake by the sarcoplasmic reticulum of malignant hyperpyrexia-susceptible pigs

Sarcoplasmic reticulum membranes were isolated from the skeletal muscle of pigs susceptible or resistant to the anaesthetic complication malignant hyperpyrexia. The skeletal muscle relaxant dantrolene was shown to be without effect on the Ca2+ -dependent ATPase activity of either sarcoplasmic reticulum preparation. The active transport of calcium by sarcoplasmic reticulum from both sources was also unaffected by the presence of dantrolene. These findings indicate that dantrolene does not exert its pharmacological action in skeletal muscle by stimulating calcium uptake by the sarcoplasmic reticulum.     

Poisoning with 2,4-dichlorophenoxyacetic acid treated by hemodialysis

In this paper four patients are presented who had been poisoned by 2,4-dichlorophenoxyacetic acid (2,4-D). The first patient, aged 51 years, had attempted to commit suicide by taking orally 400 ml of a 40% solution of 2,4-D. He was admitted in a coma, 6.5 h after poisoning. Extracorporal hemodialysis was performed and the course of the illness was satisfactory. The second patient, aged 80 years, had accidentally drunk 100 ml of a 40% solution of 2,4-D. He was admitted in a coma a few hours after poisoning. Hemodialysis and resin hemoperfusion were performed and the course of the illness was satisfactory. Prior to the above therapy the patient had a 2,4-D serum concentration of 177 mg/100 ml. 2,4-D clearance was 56,3 ml/min during this therapy. The third patient, aged 24 years, had drunk 200 ml of a 40% solution of 2,4-D in a suicide attempt, and paraquat poisoning was also suspected. He was admitted 10 h after poisoning and immediately hemodialysis and hemoperfusion were carried out: the course of the illness was satisfactory. On admittance the concentration of 2,4-D in serum was 122.5 mg/100 ml, and clearance was 72.9 ml/min during treatment.The fourth patient, aged 50 years, had accidentally drunk 100–200 ml of a 40% solution of 2,4-D. He was admitted in a coma 3 h after poisoning. Hemodialysis was performed and the course of the illness was satisfactory. On admittance the concentration of 2,4-D in serum was 37 mg/100 ml and clearance was 68.7 ml/min. On the fourth day after admittance clinical signs of 2,4-D poisoning appeared again, with a serum concentration of 43.9 ml/100 ml. The patient's condition improved after further hemodialysis. In the first two patients there was a prolonged corrected Q-T interval during coma.