Embryonic abnormalities and genotoxicity induced by 2,4-dichlorophenoxyacetic acid during indirect somatic embryogenesis in Coffea.

Indirect somatic embryogenesis (ISE) is a morphogenetic pathway in which somatic cells form callus and, later, somatic embryos (SE). 2,4-dichlorophenoxyacetic acid (2,4-D) is a synthetic auxin that promotes the proliferation and dedifferentiation of somatic cells, inducing the ISE. However, 2,4-D can cause genetic, epigenetic, physiological and morphological disorders, preventing the regeneration and/or resulting abnormal somatic embryos (ASE). We aimed to evaluate the toxic 2,4-D effect during the Coffea arabica and C. canephora ISE, assessing the SE morphology, global 5-methylcytosine levels (5-mC%) and DNA damage. Leaf explants were inoculated in media with different 2,4-D concentrations. After 90 days, the friable calli were transferred to the regeneration medium, and the number of normal and abnormal SE was monthly counted. The increase of the 2,4-D concentration increased the number of responsive explants in both Coffea. At 9.06, 18.08 and 36.24 µM 2,4-D, C. arabica presented the highest values of responsive explants, differing from C. canephora. Normal and abnormal SE regeneration increased in relation to the time and 2,4-D concentration. Global 5-mC% varied at different stages of the ISE in both Coffea. Furthermore, the 2,4-D concentration positively correlated with global 5-mC%, and with the mean number of ASE. All ASE of C. arabica and C. canephora exhibited DNA damage and showed higher global 5-mC%. The allotetraploid C. arabica exhibited greater tolerance to the toxic effect of 2,4-D than the diploid C. canephora. We conclude that synthetic 2,4-D auxin promotes genotoxic and phytotoxic disorders and promotes epigenetic changes during Coffea ISE.

Cytotoxic and genotoxic effects induced by associated commercial glyphosate and 2,4-D formulations using the Allium cepa bioassay.

Studies assessing the toxicity of glyphosate and 2,4-dichlorophenoxyacetic acid mixture are scarce. The aim of this study was to evaluate the cytotoxicity and genotoxicity of the mixture of these herbicides using Allium cepa. Roots were exposed to glyphosate (1.56 and 11.66 mg mL-1), 2,4-D (0.28 and 17.5 mg mL-1) and mixture for 24 h, based on the average concentration applied in the field and the acute reference dose (ARfD) established in Brazil. Both isolated and associated herbicides induced a significative decrease in mitotic index (MI) (P < 0.0001) in all tested concentrations. Regarding the genotoxicity results, 2,4-D and the mixture showed, at concentrations applied in the field, a significative increase of chromosomal anomalies (CA) index compared to control (P < 0.0001) and glyphosate (P = 0.024 and P = 0.0002, respectively). All tested groups from the ARfD showed a significative difference compared to the control group (P < 0.0001), as well as glyphosate and 2,4-D isolated compared to the mixture (P = 0.0005 and P < 0.0001, respectively). The most observed CA were apoptotic bodies, giant cells, and nuclear erosions. We emphasize the need for further studies assessing the toxicity of these herbicides' mixture due to the distinct effects caused in different organisms.

2,4-dichlorophenoxyacetic acid induces ROS activation in NLRP3 inflammatory body-induced autophagy disorder in microglia and the protective effect of Lycium barbarum polysaccharide.

The pesticide 2,4-dichlorophenoxyacetic acid (2,4-D) exerts neurotoxic effects; however, its action mechanism remains unclear. Here, we used BV2 cells as a model and divided them into six groups: control group (serum-free medium), lipopolysaccharide (LPS) (1 µg/mL), 2,4-D (1.2 µmol/mL), Lycium barbarum polysaccharide (LBP; 300 µg/mL LBP), LPS (1 µg/mL) + LBP (300 µg/mL), and 2,4-D (1.2 µmol/mL) + LBP (300 µg/mL) with dimethyl sulfoxide as the solvent. Our results showed that 2,4-D treatment decreased superoxide dismutase and glutathione peroxidase activities and increased malondialdehyde content. The percentage of microglial activation (co-expression of ionized calcium-binding adaptor protein-1 + CD68) in the LPS and 2,4-D groups and the levels of tumor necrosis factor alpha, interleukin (IL) 1 beta, IL-6, and IL-18 in the cell supernatant were increased. The protein and mRNA levels of Nod-like receptor protein 3 (NLRP3), apoptosis-associated speck-like protein, caspase-1, IL-1ß, IL-18, and p62 increased, whereas those of LC3II/I and Beclin-1 decreased in the 2,4-D group. The protein expression and mRNA levels of NLRP3, cleaved caspase-1, IL-1ß, IL-18, and p62 decreased significantly, whereas the protein expression and mRNA levels of LC3II/I and Beclin-1 increased in small interfering RNA of NLRP3-treated BV2 cells stimulated with 2,4-D and LPS. In conclusion, 2,4-D enhanced cell migration, promoted oxidative stress, induced excessive release of mitochondrial reactive oxygen species, promoted microglial cell activation, released inflammatory factors, activated NLRP3 inflammasomes, and inhibited autophagy. Meanwhile, LBP reduced inflammation and the release of mitochondrial reactive oxygen species, inhibited NLRP3 inflammasome activation, and regulated autophagy, thereby playing a neuroprotective role.

Protective effect of Thymus munbyanus aqueous extract against 2,4-dichlorophenoxyacetic acid-induced nephrotoxicity in Wistar rats.

The aim of the present study was to examine the possible protective effects of the aqueous extract of Thymus munbyanus (TMAE) against 2,4-dichlorophenoxyacetic acid (2,4-D)-induced oxidative stress and renal injury in the kidney of male albino rats. Furthermore, TMAE was assessed to determine the phenolic content. In vitro, antioxidant activities were evaluated by DPPH radical scavenging, inhibition of ß-carotene bleaching, and reducing power. The results showed that TMAE contained high amounts of phenolics, flavonoids, and tannins. Additionally, 24 rats were randomly divided into four groups: a control group, and three groups treated with TMAE (10 mL/kg body weight), 2,4-D (5 mg/kg body weight), and 2,4-D/combined with the TMAE for 4 weeks. Treatment with 2,4-D induced kidney dysfunctions demonstrated as an increase in the potential markers of renal filtration, namely urea and creatinine, associated with a decrease in uric acid. Moreover, 2,4-D increased malondialdehyde and carbonyl protein levels. Additionally, reduced glutathione (GSH) content, as well as superoxide dismutase (SOD), glutathione peroxidase (GPx), and glutathione S-transferase (GST) activities were significantly decreased. Our results demonstrated that oral TMAE supplementation in 2,4-D-treated rats improved and restored some blood parameters and alleviated the adverse cytotoxic effects of 2,4-D by increasing certain antioxidants, consequently attenuating the intensity of oxidative stress induced by 2,4-D; this was confirmed by the histological improvements observed in the kidneys. In conclusion, TMAE demonstrated potential as a natural antioxidant, effectively alleviating 2,4-D induced kidney oxidative injury.

Elevated pulmonary levels of Axin2 in mice exposed to herbicide 2,4-D with or without endotoxin.

2,4-Dichlorophenoxyacetic acid (2,4-D), a member of the phenoxy family of herbicides is commonly used in agriculture for controlling broadleaf weeds but its uncontrolled and incoherent use has been linked to incidences of lung toxicity. The present study aimed to understand the molecular mechanisms behind the 2,4-D alone or in combination with endotoxin (lipopolysaccharide [LPS]) induced pulmonary toxicity. Blood and lung samples were collected from Swiss albino mice (n = 48) following chronic exposure to high (37 mg/kg; 1/10th of LD50 ) and low (18.5 mg/kg; 1/20th of LD50 ) doses of 2,4-D alone or in combination with endotoxin (80 µg/animal). Transcriptome analysis revealed Wnt Canonical signaling as one of the top dysregulated pathways in mice lung following exposure to 2,4-D with and without endotoxin (LPS) co-exposure. Global view of differentially expressed genes showed increased messenger RNA expression of Axin2 by 0.26, 2.58, 3.14, 2.59, and 2.97 folds following exposure to LPS, high dose alone or in combination with LPS and low dose alone or in combination with LPS, respectively. The microarray data were validated using quantitative polymerase chain reaction and immunohistochemistry. Furthermore, the plasma concentration of Axin2 was elevated in the high dose group as revealed by Sandwich ELISA. The data taken together suggest a role of Axin2 to activate the Canonical Wnt signaling pathway in 2,4-D and or endotoxin-induced lung damage in mice.

Effects of 2,4-dichlorophenoxyacetic acid on the expression of NLRP3 inflammasome and autophagy-related proteins as well as the protective effect of Lycium barbarum polysaccharide in neonatal rats.

The pesticide 2,4-dichlorophenoxyacetic acid (2,4-D) has neurotoxic effects, but its mechanism is not clear. In this study, a 2,4-D (75 mg/kg. b.w) exposure model was established in SD rats with colostrum. Lipopolysaccharide (1 mg/kg b.w) was used as the positive control, and Lycium barbarum polysaccharide (LBP, 50 mg/kg b.w) was used as an intervention factor to explore the neurotoxic effect of 2,4-D and the neuroprotective effect of LBP. Our research results show that 2,4-D causes a decrease in the number of hippocampal CA3 pyramidal cells and pyknosis in nuclei with a triangular or irregular shape and that rats show signs of anxiety or depression. In rat serum, superoxide dismutase, and glutathione peroxidase activity decreased, while malondialdehyde content increased. Protein and mRNA levels of TNFα, IL-6, IL-1ß, IL-18, NLRP3, ASC, caspase-1, IL-1ß, IL-18, and p62 increased, while those of LC3-II/LC3-I and Beclin-1 decreased in hippocampal tissues. In conclusion, 2,4-D increased the oxidative stress level, induced neuroinflammatory response, and decreased the autophagy level in experimental rats. LBP may have upregulated the autophagy level in the body by inhibiting the activation of the NLRP3 inflammasome, thus playing a neuroprotective role.

2,4-Dichlorophenoxyacetic acid induced hepatic and renal toxicological perturbations in rat model: Attenuation by selenium supplementation.

2,4-Dichlorophenoxyacetic acid (2,4-D) is a commercially used herbicide to manage broadleaf weeds that have various toxicological and ecological effects. In view of ever-escalating use of 2,4-D, risk assessment becomes mandatory to ensure the safety of both human health and the ecosystem. Oxidative injury has been expected as a possible mechanism implicated in 2,4-D toxicity. The present study was planned and conducted to explore the antioxidant potential of selenium (Se) supplementation to moderate the 2,4-D hepatic and renal toxicity in a rat model. The rats were randomly assigned to four equal groups and treated via oral gavage for a period of 4 weeks. Group I: received deionized water as a vehicle, group II: received 2,4-D (150 mg-1 kg-1 day-1), group III: received Se supplement (1 mg-1 kg-1 day-1), and group IV: received 2,4-D (150 mg-1 kg-1 day-1) and Se supplement (1 mg-1 kg-1 day-1) simultaneously. After 4 weeks of administration, 2,4-D induced toxicity was observed, as manifested by disrupted levels of plasma urea, creatinine, alkaline phosphatase (ALP), aspartate aminotransferase (AST), and alanine aminotransferase (ALT). Further, 2,4-D caused a considerable increase in tissue malondialdehyde (MDA) levels and decreased activity of antioxidant enzymes, including superoxide dismutase, catalase, and glutathione reductase. Se supplementation exhibited its antioxidant properties by significantly improving urea, creatinine, ALP, AST, and ALT, and MDA levels and antioxidant enzyme activities. In conclusion, the results suggest that 2,4-D induced hepatic and renal toxicities were attenuated by Se supplementation probably owing to its antioxidant properties.

Magnesium supplementation ameliorates toxic effects of 2,4-dichlorophenoxyacetic acid in rat model.

2,4-Dichlorophenoxyacetic acid (2,4-D) is an extensively used herbicide in the field of agriculture, its ever-escalating use induces toxicity, health effects, and environmental impact. Oxidative stress plays a key role in pathogenesis of 2,4-D-induced liver and kidney damage. Magnesium (Mg) is a highly effective antioxidant agent in restoring oxidative damage by directly influencing the metabolic and physiological processes. Therefore, the present study aimed to evaluate Mg role in ameliorating the oxidative damages provoked by 2,4-D in rat model. Male Wistar rats (180-220 g) were distributed into four groups and treated intragastrically for 4 weeks. Group 1: control, group 2: 2,4-D (150 mg/kg body weight/day), group 3: simultaneously treated with 2,4-D (150 mg/kg body weight/day) and Mg supplement (50 mg/kg body weight/day), and group 4: Mg supplement (50 mg/kg body weight/day). Under experimental conditions, plasma hepatic and renal biomarkers, tissue oxidative status, and antioxidant enzymes activities were investigated. Results demonstrated that 2,4-D intoxication caused hepatic and renal impairments as indicated by the significantly increased (p < 0.001) alkaline phosphatase, alanine aminotransferase, aspartate aminotransferase, urea, creatinine, and blood urea nitrogen levels. In addition, 2,4-D caused a significant enhancement (p < 0.001) in the level of malondialdehyde as well as reduction (p < 0.001) of the superoxide dismutase, catalase, and glutathione reductase activities in both hepatic and renal tissues. Mg treatment prevented and reversed the toxic variations induced by 2,4-D. In general, these outcomes suggest that Mg may have antioxidant potential and ameliorative effects against 2,4-D provoking hepatic and renal toxicity in rat model.

Potential Role of Selenium Against Hepatotoxicity Induced by 2,4-Dichlorophenoxyacetic Acid in Albino Wistar Rats.

The present study aims to investigate the hepatoprotective effects of selenium on toxicity induced by 'Désormone Lourd' based on 2,4-dichlorophenoxyacetic acid in Wistar rats. Male Wistar rats were divided into four groups and were treated orally. The (C) group was used as a control, while the test groups were treated with Se (0.2 mg/kg b.w.), 2,4-D (5 mg/kg b.w.) or both (2,4-D + Se) for 4 weeks. Our results showed that chronic treatment with 2,4-D resulted in hepatotoxicity, as revealed by an increase in liver function markers Aminotransferases (ALT, AST), alkaline phosphatase (ALP), lactate dehydrogenase (LDH) and total bilirubin (TB), along with reduced total protein content and albumin. An overall pro-oxidant effect was associated with a decrease in the reduced glutathione (GSH) content and the enzymatic activity of glutathione-S-transferase (GST), catalase (CAT), superoxide dismutase (SOD) and glutathione peroxidase (GPx), and an increase in malondialdehyde (MDA) and protein carbonyl levels (PCO). Microscopic observation of liver in 2,4-D-treated rats reveals lesions, which results in perivascular inflammatory infiltration around the vessel, sinusoidal dilatation and vacuolization of hepatocytes. However, selenium supplementation in 2,4-D-treated rats elicited a reduction in the toxic effects of the pesticide by improving the studied parameters, which was confirmed by the histological study of the liver. Selenium appears to have a promising prophylactic effect through its effective anti-radical action against the hepatotoxic effects of 2,4-D.

Modulation of physiological responses with TiO2 nano-particle in Azolla pinnata R.Br. under 2,4-D toxicity.

The present work is emphasised with the herbicidal tolerance of Azolla pinnata R.Br. and its modulation with TiO2 nano-particle. Both carbohydrate and nitrogen metabolism were effected with 2,4-D as herbicide and in few cases TiO2-NP had recovered few detrimental effects. From the nutrient status in Azolla it recorded the recovery of nitrogen as well as potassium by TiO2-NP but not in case of phosphorus. However, a conversion of nitrate to ammonium was more induced by TiO2-NP under herbicidal toxicity. Similar results were obtained for inter-conversion of amino acid-nitrate pool, but no changes with glutamine synthase activity with TiO2-NP. Initially, the effects of 2,4-D was monitored with changes of chlorophyll content but had not been recovered with nanoparticle. Photosynthetic reserves expressed as both total and reducing sugar were insensitive to TiO2-NP interference but activity of soluble and wall bound invertase was in reverse trend as compared to control. The 2,4-D mediated changes of redox and its oxidative stress was ameliorated in plants with over expressed ADH activity. As a whole the Azolla bio system with TiO2 supplementation may be useful in sustenance against 2,4-D toxicity through recovery of nitrogen metabolism. Thus, Azolla-TiO2-NP bio system would be realised to monitor the herbicidal toxicity in soil and its possible bioremediation.

Weight-of-the-evidence evaluation of 2,4-D potential for interactions with the estrogen, androgen and thyroid pathways and steroidogenesis.

A comprehensive weight-of-the-evidence evaluation of 2,4-dichlorophenoxyacetic acid (2,4-D) was conducted for potential interactions with the estrogen, androgen and thyroid pathways and with steroidogenesis. This assessment was based on an extensive database of high quality in vitro, in vivo ecotoxicological and in vivo mammalian toxicological studies. Epidemiological studies were also considered. Toxicokinetic data provided the basis for determining rational cutoffs above which exposures were considered irrelevant to humans based on exceeding thresholds for saturation of renal clearance (TSRC); extensive human exposure and biomonitoring data support that these boundaries far exceed human exposures and provide ample margins of exposure. 2,4-D showed no evidence of interacting with the estrogen or androgen pathways. 2,4-D interacts with the thyroid axis in rats through displacement of thyroxine from plasma binding sites only at high doses exceeding the TSRC in mammals. 2,4-D effects on steroidogenesis parameters are likely related to high-dose specific systemic toxicity at doses exceeding the TSRC and are not likely to be endocrine mediated. No studies, including high quality studies in the published literature, predict significant endocrine-related toxicity or functional decrements in any species at environmentally relevant concentrations, or, in mammals, at doses below the TSRC that are relevant for human hazard and risk assessment. Overall, there is no basis for concern regarding potential interactions of 2,4-D with endocrine pathways or axes (estrogen, androgen, steroidogenesis or thyroid), and thus 2,4-D is unlikely to pose a threat from endocrine disruption to wildlife or humans under conditions of real-world exposures.

Oxidative stress and metabolic perturbations in Escherichia coli exposed to sublethal levels of 2,4-dichlorophenoxyacetic acid.

The chlorophenoxy herbicide 2,4-dichlorophenoxyacetic acid (2,4-D) is used extensively worldwide despite its known toxicity and our limited understanding of how it affects non-target organisms. Escherichia coli is a suitable model organism to investigate toxicity and adaptation mechanisms in bacteria exposed to xenobiotic chemicals. We developed a methodical platform that uses atomic force microscopy, metabolomics and biochemical assays to quantify the response of E. coli exposed to sublethal levels of 2,4-D. This herbicide induced a filamentous phenotype in E. coli BL21 and a similar phenotype was observed in a selection of genotypically diverse E. coli strains (A0, A1, B1, and D) isolated from the environment. The filamentous phenotype was observed at concentrations 1000 times below field levels and was reversible upon supplementation with polyamines. Cells treated with 2,4-D had more compliant envelopes, significantly remodeled surfaces that were rougher and altered vital metabolic pathways including oxidative phosphorylation, the ABC transport system, peptidoglycan biosynthesis, amino acid, nucleotide and sugar metabolism. Most of the observed effects could be attributed to oxidative stress, consistent with increases in reactive oxygen species as a function of 2,4-D exposure. This study provides direct evidence that 2,4-D at sublethal levels induces oxidative stress and identifies the associated metabolic changes in E. coli.

Post-Vietnam military herbicide exposures in UC-123 Agent Orange spray aircraft.

BACKGROUND: During the Vietnam War, approximately 20 million gallons of herbicides, including ~10.5 million gallons of dioxin-contaminated Agent Orange, were sprayed by about 34 UC-123 aircraft that were subsequently returned to the United States, without decontamination or testing, to three Air Force reserve units for transport operations (~1971-1982). In 1996, observed dioxin contamination led to withdrawal of these UC-123s from public auction and to their smelting in 2009. Current Air Force and Department of Veterans Affairs policies stipulate that "dried residues" of chemical herbicides and dioxin had not lead to meaningful exposures to flight crew and maintenance personnel, who are thus ineligible for Agent Orange-related benefits or medical examinations and treatment. Sparse monitoring data are available for analysis. METHODS: Three complementary approaches for modeling potential exposures to dioxin in the post-Vietnam war aircraft were employed: (1) using 1994 and 2009 Air Force surface wipe data to model personnel exposures and to estimate dioxin body burden for dermal-oral exposure for dried residues using modified generic US Environmental Protection Agency intake algorithms; (2) comparing 1979 Air Force 2,4- dichlorophenoxyacetic acid and 2,4-5-trichlorophenoxyacetic acid air samples to saturated vapor pressure concentrations to estimate potential dioxin exposure through inhalation, ingestion and skin contact with contaminated air and dust; and (3) applying emission models for semivolatile organic compounds from contaminated surfaces to estimate airborne contamination. RESULTS: Model (1): Body-burden estimates for dermal-oral exposure were 0.92 and 5.4pg/kg body-weight-day for flight crew and maintainers. The surface wipe concentrations were nearly two orders of magnitude greater than the US Army guidance level. Model (2): measured airborne concentrations were at least five times greater than saturated vapor pressure, yielding dioxin estimates that ranged from 13.2-27.0pg/m(3), thus supporting the likelihood of dioxin dust adsorption. Model (3): Theoretical models yielded consistent estimates to Model 2, 11-49pg/m(3), where the range reflects differences in experimental value of dioxin vapor pressure and surface area used. Model (3) results also support airborne contamination and dioxin dust adsorption. CONCLUSIONS: Inhalation, ingestion and skin absorption in aircrew and maintainers were likely to have occurred during post-Vietnam use of the aircraft based on the use of three complementary models. Measured and modeled values for dioxin exceeded several available guidelines. Deposition-aerosolization-redeposition homeostasis of semivolatile organic compound contaminants, particularly dioxin, is likely to have continually existed within the aircraft. Current Air Force and Department of Veterans Affairs policies are not consistent with the available industrial hygiene measurements or with the widely accepted models for semivolatile organic compounds.

An F1-extended one-generation reproductive toxicity study in Crl:CD(SD) rats with 2,4-dichlorophenoxyacetic acid.

2,4-Dichlorophenoxyacetic acid (2,4-D) was assessed for systemic toxicity, reproductive toxicity, developmental neurotoxicity (DNT), developmental immunotoxicity (DIT), and endocrine toxicity. CD rats (27/sex/dose) were exposed to 0, 100, 300, 600 (female), or 800 (male) ppm 2,4-D in diet. Nonlinear toxicokinetic behavior was shown at high doses; the renal clearance saturation threshold for 2,4-D was exceeded markedly in females and slightly exceeded in males. Exposure was 4 weeks premating, 7 weeks postmating for P1 males and through lactation for P1 females. F1 offspring were examined for survival and development, and at weaning, pups were divided in cohorts, by sex and dose, and by systemic toxicity (10), DNT (10), DIT (20), and reproductive toxicity (≥ 23). Remaining weanlings were evaluated for systemic toxicity and neuropathology (10-12). Body weight decreased during lactation in high-dose P1 females and in F1 pups. Kidney was the primary target organ, with slight degeneration of proximal convoluted tubules observed in high-dose P1 males and in high-dose F1 males and females. A slight intergenerational difference in kidney toxicity was attributed to increased intake of 2,4-D in F1 offspring. Decreased weanling testes weights and delayed preputial separation in F1 males were attributed to decreased body weights. Endocrine-related effects were limited to slight thyroid hormone changes and adaptive histopathology in high-dose GD 17 dams seen only at a nonlinear toxicokinetic dose. 2,4-D did not cause reproductive toxicity, DNT, or DIT. The "No Observed Adverse Effect Level" for systemic toxicity was 300 ppm in both males (16.6 mg/kg/day) and females (20.6 mg/kg/day), which is approximately 6700- to 93 000-fold higher than that reported for 2,4-D exposures in human biomonitoring studies.

2,4-D exposure and risk assessment: comparison of external dose and biomonitoring based approaches.

Conventional chemical exposure assessment relies upon measurements or estimates of chemical concentrations in environmental media, food, or products, in combination with assumptions regarding contact rates, in order to estimate external doses (ppm in air) or intake rates of chemicals (e.g., mg/kg/day ingested). A risk assessment is conducted by comparing these external or intake dose estimates to appropriate (e.g., route-specific) exposure guidance values (e.g., Reference Dose or Reference Concentration) to assess whether exposures are exceeding levels of concern. Human biomonitoring, in which concentrations of chemicals are measured in blood or urine, is being increasingly used as an alternative or complementary exposure assessment. The Biomonitoring Equivalent, which is the translation of a Reference Dose to an equivalent concentration of a compound in blood or urine, provides a parallel means to interpret biomonitoring data in order to assess whether chemical-specific exposures exceed levels of concern. This manuscript presents a side-by-side comparison of the two approaches for assessing exposures and risks for a case study compound, 2,4-dichlorophenoxyacetic acid (2,4-D). The findings from this case study indicate that the external dose-based assessments result in estimates of exposure and resulting hazard quotients that are consistently several-fold higher than those based on biomonitoring data. These comparisons support a conclusion that exposure assessments conducted as part of the registration process for 2,4-D incorporate sufficiently conservative assumptions.

Uncaria tomentosa extracts protect human erythrocyte catalase against damage induced by 2,4-D-Na and its metabolites.

The effect of ethanolic and aqueous extracts from leaves and bark of Uncaria tomentosa was studied, with particular attention to catalase activity (CAT - EC. 1.11.1.6). We observed that all tested extracts, at a concentration of 250 µg/mL were not toxic to erythrocyte catalase because they did not decreased its activity. Additionally, we investigated the protective effect of extracts on changes in CAT activity in the erythrocytes incubated with sodium salt of 2,4-dichlorophenoxyacetic acid (2,4-D-Na) and its metabolites i.e., 2,4-dichlorophenol (2,4-DCP) and catechol. Previous investigations showed that these chemicals decreased activity of erythrocyte catalase (Bukowska et al., 2000; Bukowska and Kowalska, 2004). The erythrocytes were divided into two portions. The first portion was incubated for 1 and 5h at 37°C with 2,4-D-Na, 2,4-DCP and catechol, and second portion was preincubated with extracts for 10 min and then incubated with xenobiotics for 1 and 5h. CAT activity was measured in the first and second portion of the erythrocytes. We found a protective effect of the extracts from U. tomentosa on the activity of catalase incubated with xenobiotics studied. Probably, phenolic compounds contained in U. tomentosa scavenged free radicals, and therefore protected active center (containing -SH groups) of catalase.

Hypolipidimic and antioxidant activities of virgin olive oil and its fractions in 2,4-diclorophenoxyacetic acid-treated rats.

OBJECTIVE: We examined the effects of extra virgin olive oil (EVOO) and its hydrophilic and lipophilic fractions on serum lipids, oxidative stress, and morphologic and functional liver damages induced by 2,4-diclorophenoxyacetic acid (2,4-D). METHODS: Male Wistar rats were divided randomly into eight groups: control; 2,4-D at a dose of 5 mg/kg of body weight (2,4-D); 2,4-D plus EVOO (2,4-D/EVOO); 2,4-D plus the hydrophilic fraction (2,4-D/OOHF); 2,4-D plus the lipophilic fraction (2,4-D/OOLF); only EVOO (EVOO); only the hydrophilic fraction (OOHF); and only the lipophilic fraction (OOLF). These components were administered daily by gavage for 4 wk. RESULTS: A hepatic architecture aberration, increased activities of aspartate and alanine aminotransferase enzymes, total and low-density lipoprotein cholesterol, and malondialdehyde (MDA) level, and a decreased antioxidant defense system were observed in the 2,4-D group. The administration of EVOO restored the damage caused by 2,4-D by a significant decrease of plasma total and low-density lipoprotein levels and a moderate increase of high-density lipoprotein cholesterol. The 2,4-D/OOHF group exhibited a pronounced enhancement of the antioxidant defense system by an increase of superoxide dismutase, catalase, and glutathione peroxidase levels and a decrease of plasma and liver MDA levels. However, less improvement in the liver histoarchitecture and antioxidant status was observed in rats supplemented with OOLF diet, despite its richness in α-tocopherol. CONCLUSION: Extra virgin olive oil may be a potential functional food source of antioxidants than can decrease the frequency of cardiovascular diseases and liver damage.

Dietary olive oil effect on antioxidant status and fatty acid profile in the erythrocyte of 2,4-D- exposed rats.

BACKGROUND: Oxidative stress produced by reactive oxygen species (ROS) has been linked to the development of several diseases such as cardiovascular, cancer, and neurodegenerative diseases. This study investigates the possible protective effect of extra virgin olive oil (EVOO), lipophilic fraction (OOLF) and hydrophilic fraction (OOHF) on oxidative stress and fatty acid profile of erythrocytes in 2,4-D treated rats. METHODS: Male Wistar rats were divided randomly into eight groups: control (C), (2,4-D) at a dose of 5 mg/kg b.w., (2,4-D/EVOO) was given 2,4-D plus EVOO, (2,4-D/OOHF) that received 2,4-D plus hydrophilic fraction, (2,4-D/OOLF) treated with 2,4-D plus lipophilic fraction, (EVOO) that received only EVOO, (OOHF) was given hydrophilic fraction and (OOLF) treated with lipophilic fraction. These components were daily administered by gavages for 4 weeks. RESULTS: 2,4-D treatment lead to decrease of antioxidant enzyme activities, namely, superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) and glutathione reductase (GR) associated with a higher amount of MDA level. Erythrocyte membranes' fatty acid composition was also significantly modified with 2,4-D exposure. EVOO and hydrophilic fraction supplemented to rats with or not 2,4-D treatment enhanced the antioxidant enzyme activities and reduced the MDA level. However, lipophilic fraction did not show any improvement in oxidative damage induced by 2,4-D in spite its richness in MUFA and vitamins. CONCLUSION: EVOO administered to 2,4-D-treated rats protected erythrocyte membranes against oxidative damage by means of preventing excessive lipid peroxidation to increase the MUFA composition and increase maintaining antioxidants enzymes at normal concentrations.

Pesticides, soft-tissue sarcoma and non-Hodgkin lymphoma--historical aspects on the precautionary principle in cancer prevention.

BACKGROUND: After the 2(nd) World War a long range of chemical agents have been introduced on the market, both in Sweden and most other countries. From the 1950's several pesticides gained increasing use in agriculture and forestry. In the 1970's public concern increased in Sweden especially regarding use of phenoxy herbicides to combat deciduous wood, although statements from different authorities were reassuring of the safety. MATERIALS AND METHODS: At the end of the 1970's the author and his colleagues published the first scientific evidence of an association between exposure to phenoxyacetic acids, chlorophenols and certain malignant tumours, i.e., soft-tissue sarcoma and malignant lymphoma. The study subjects were also exposed to contaminating dioxins such as 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). Later studies showed also an association between certain persistent organic pollutants such as polychlorinated biphenyls and non-Hodgkin lymphoma (NHL) with an interaction with titers of antibodies to Epstein-Barr virus early antigen. These results have been corroborated in other studies. DISCUSSION: Over the years industry and its allied experts have attacked our studies, but in 1997 IARC classified TCDD as a human carcinogen, Group I. The increasing incidence of NHL in Sweden levelled off about 1990. The author postulated that the regulation or ban of the use of chlorophenols, certain phenoxy herbicides and some persistent organic pollutants in Sweden back in the 1970s has contributed to the now decreasing incidence of NHL. Unfounded criticism from industry experts may prohibit the precautionary principle and early warnings of cancer risk can be ignored. Cancer risks by certain chlorinated phenols may serve as a model of how the precautionary principle should be used by taking early warnings seriously.