Administration Of Acrylamide Research Articles

Prophylaxis with Bacopa monnieri Attenuates Acrylamide Induced Neurotoxicity and Oxidative Damage via Elevated Antioxidant Function

Acrylamide (ACR) is a water-soluble, vinyl monomer that has multiple chemical and industrial applications. Exposure to ACR causes neuropathy and associated neurological defects including gait abnormalities and skeletal muscle weakness, due to impaired neurotransmitter release and eventual neurodegeneration. Using in vivo and in vitro models, we examined whether oxidative events are involved in ACR-mediated neurotoxicity and whether these could be prevented by natural plant extracts. Administration (i.p.) of ACR in mice (40 mg/kg bw/ d for 5d) induced significant oxidative damage in the brain cortex and liver as evidenced by elevated lipid peroxidation, reactive oxygen species and protein carbonyls. This was associated with lowered antioxidant activities including antioxidant enzymes (catalase, glutathione-s-transferase) and reduced glutathione (GSH) compared to untreated controls. Similarly, exposure of N27 neuronal cells in culture to ACR (1-5 mM) caused dose-dependent neuronal death and lowered GSH. Interestingly, dietary supplementation with the leaf powder of Bacopa monnieri (BM) (which possesses neuroprotective properties and nootropic activity) in mice for 30 days offered significant protection against ACR toxicity and oxidative damage in vivo. Similarly, pretreatment with BM protected the N27 cells against ACR-induced cell death and associated oxidative damage. Co-treatment and pre-treatment of Drosophila melanogaster with BM extract protected against ACR-induced locomotor dysfunction and GSH depletion. We infer that BM displays prophylactic effects against ACR induced oxidative damage and neurotoxicity with potential therapeutic application in human pathology associated with neuropathy.

Role of coenzyme Q10 in testicular damage induced by acrylamide in weaned albino rats

Introduction Acrylamide is a vinyl monomer frequently used in the polymer industry. It has the potential to adversely affect male reproductive capacity. Coenzyme Q10 (CoQ10) is a strong antioxidant. Objectives The objectives of this study were to examine the histological changes in the testis after the administration of acrylamide and the possible protective role of CoQ10. Materials and methods Thirty weaned male albino rats aged 21 days were classified into three groups of10 rats each: group I was the control group; group II (acrylamide-treated rats) received oral acrylamide at 15 mg/kg body weight/day; and group III (protected group) received both acrylamide (at the same previous dose) and an intraperitoneal injection of CoQ10 at 10 mg/kg body weight/day. After 8 weeks, blood samples were taken to measure the serum testosterone level. The testes from each animal were dissected out and processed for light microscope examination using H&E stains and immunohistochemical stains for the detection of proliferating cell nuclear antigen and inducible nitric oxide synthase. Morphometric and statistical analyses were carried out. Results After the administration of acrylamide, variable degrees of tubular affection were observed. Some tubules were shrunken with disorganized germinal epithelium. Spermatogonia contained darkly stained nuclei. Cellular vacuolations as well as sloughed spermatogenic cells into the lumen were observed. The interstitium was widened with interstitial hyperplasia, eosinophilic material, and congestedcapillaries. Immunohistochemically, acrylamide treatment induced a markedreduction in the number of proliferating cell nuclear antigen immunoreactive spermatogonia and spermatocytes and an increase in the number of inducible nitric oxide synthase immunoreactive spermatids and spermatocytes. The concomitant administration of CoQ10 with acrylamide induced an observable protectionagainst these changes. Conclusion CoQ10 played a protective role against acrylamide-induced testicular damage.

Acrylamide-mediated subacute testicular and genotoxicity, is it reversible?

Background and aim Humans may be susceptible to acrylamide poisoning that may induce genotoxicity and testicular toxicity. Studies on recovery from acrylamide-induced reproductive toxicity are scarce. This study aimed to evaluate the natural recovery of testicular and genotoxicity caused by subacute exposure to acrylamide in rats. Materials and methods Thirty-six male Sprague–Dawley rats were divided into six acrylamide-treated groups and one control group. They were dosed orally once a day with acrylamide at 45 mg/kg/day for 5 days and then held for different recovery periods (3, 5, 10, 15, 25, and 55 days). The parameters examined were sperm count and morphology, hormonal assay, histopathology, comet assay, and apoptosis detection in testis. Statistical analysis was carried out by one-way analysis of variance using the SPSS software (significance was considered at P<0.05). Results Sperm abnormalities (detached heads and intersegmentation) detected were decreased markedly after 55 days of recovery. Transforming growth factor-β1 level was significantly increased following 10 days of recovery and then reverted to control levels. Abnormalities in the testis were detected between 3 and 25 days of recovery. None of these abnormalities was observed after 55 days of recovery besides a reduction in the luminal sperm reserve of the epididymis. Significant DNA damage was also induced by arylamide in blood leukocytes and reached its maximum after 3 days of recovery (60%). A gradual reduction in cell damage and tail moment occurred until 15 days of recovery (1%). Conclusion This study showed that oral administration of acrylamide in rats at a dose of 45 mg/kg for 5 consecutive days induced systemic effects as well as reproductive toxicity. In general, these effects were reversible, although the extent of the recovery during the 55-day period studied ranged from complete to partial.

Hazardous effects of acrylamide on immature male and female rats

Acrylamide (ACR) is an industrial chemical which induces neurotoxic effects in experimental animals and humans. The present study was carried out to investigate the hematological, biochemical, neurological and histopathological effects of ACR on immature male and female rats. Animals were divided into 2 main groups; immature male group and immature female group and all rats were treated for 28 consecutive days. Each main group subsequently was divided into 2 subgroups: (I) Untreated control group that received a daily oral administration of distilled water and (II) ACR treated rats which received a daily oral administration of ACR (15 mg/kg/body weight). The results obtained indicate that ACR administration induced some behavioral disorders in the movement of immature male and female rats as well as loss of body weight. ACR induced a significant decrease in hemoglobin (Hb), erythrocytes (RBCS), hematocrit (HCT) and lymphocyte levels of young female rats. ACR significantly increased serum glucose, total cholesterol and triglycerides concentrations of both immature male and female rats. While, significant increase in the total urea concentration was noticed only in the immature male rats following ACR administration. Moreover, ACR induced marked increase in the activities of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) in the immature male and female rats. On the other hand, the activities of serum alkaline phosphatase (ALP) and acetylcholinesterase (AChE) were significantly decreased in both treated groups. ACR caused a significant increase in norepinephrin (NE), glutamate, aspartate and taurine, while it reduced dopamine (DA) and serotonin (5-HT) levels. In conclusion, the present study showed that, ACR induced hazardous effects on immature male and female rats. So, we recommended that children must avoid fast or junk foods. Key words: Acrylamide, hematological, biochemical, neurological. histopathological, immature.

Reduced BACE1 Activity Enhances Clearance of Myelin Debris and Regeneration of Axons in the Injured Peripheral Nervous System

β-Site amyloid precursor protein (APP) cleaving enzyme 1 (BACE1) is an aspartyl protease best known for its role in generating the amyloid-β peptides that are present in plaques of Alzheimer's disease. BACE1 has been an attractive target for drug development. In cultured embryonic neurons, BACE1-cleaved N-terminal APP is further processed to generate a fragment that can trigger axonal degeneration, suggesting a vital role for BACE1 in axonal health. In addition, BACE1 cleaves neuregulin 1 type III, a protein critical for myelination of peripheral axons by Schwann cells during development. Here, we asked whether axonal degeneration or axonal regeneration in adult nerves might be affected by inhibition or elimination of BACE1. We report that BACE1 knock-out and wild-type nerves degenerated at a similar rate after axotomy and to a similar extent in the experimental neuropathies produced by administration of paclitaxel and acrylamide. These data indicate N-APP is not the sole culprit in axonal degeneration in adult nerves. Unexpectedly, however, we observed that BACE1 knock-out mice had markedly enhanced clearance of axonal and myelin debris from degenerated fibers, accelerated axonal regeneration, and earlier reinnervation of neuromuscular junctions, compared with littermate controls. These observations were reproduced in part by pharmacological inhibition of BACE1. These data suggest BACE1 inhibition as a therapeutic approach to accelerate regeneration and recovery after peripheral nerve damage.

Effects of oral acrylamide intake on blood viscosity parameters in rats

Acrylamide which is formed via reaction of reducing sugars with amino acids during food processing at high temperatures is not only neurotoxic and carcinogenic, but it also damages erythrocyte membrane and generates micronucleated erythrocytes. In the present study, effects of chronic administration of acrylamide at a dose which does not induce neurotoxicity were evaluated on blood viscosity parameters (hematocrit, erythrocyte deformability, erythrocyte aggregation and plasma viscosity). Twenty adult male Sprague-Dawley rats were divided into control and acrylamide groups. The acrylamide group received 10 mg/kg/day acrylamide, whereas the control group received saline (vehicle), both in 10 ml/kg/day volume via gastric gavage. Erythrocyte aggregation and deformability were measured with LORCA and plasma viscosity with cone-plate viscometer. Erythrocyte deformability was measured before, and at the end of the 3rd and the 5th weeks of acrylamide administration. Hematocrit, erythrocyte aggregation and plasma viscosity were measured only at the end of the 5th week. Acrylamide caused a significant decrease in the deformability index of erythrocytes (at the end of the 3rd week, control: 0.606 ± 0.003, acrylamide: 0.595 ± 0.003, p < 0.05) (at the end of the 5th week, control: 0.606 ± 0.002, acrylamide: 0.588 ± 0.002, p < 0.01). Aggregation tendency and plasma viscosity were slightly higher in the acrylamide group, however the difference was not statistically significant. These results imply that acrylamide which does not cause neurotoxicity in rats may alter blood viscosity if chronically taken.

Potent neuroprotective role of novel melatonin derivatives for management of central neuropathy induced by acrylamide in rats

Acrylamide (ACR) has been shown to be a neurotoxic agent for both laboratory animals and human. The present study aimed at synthesizing new functionalized melatonin derivatives bearing promising heterocyclic moiety that could be expected to have protective effect against ACR-induced neurotoxicity in adult female rats. The novel melatonin derivatives 4, 6, 7 and 11 were synthesized and their chemical structures were confirmed by studying their analytical and spectral data. The administration of ACR [i.p., 50 mg kg −1 body weight (b. wt.)] alone resulted in significant increase in brain malondialdehyde level (MDA) and lactate dehydrogenase (LDH) activity whereas it caused significant decrease in brain monoamines levels and antioxidant enzymes activity. Treatment with melatonin derivatives 4, 6, 7 and 11 (i.p., 50 mg kg −1 b. wt) prior to ACR produced significant decrease in brain MDA level and LDH activity with concomitant significant increase in brain monoamines and antioxidant enzymes activity. It could be concluded that the new synthesized melatonin derivatives exhibited promising protective activity against ACR-induced neurotoxicity.

Lack of modifying effects of prepubertal exposure to acrylamide (AA) on N-methyl-N-nitrosourea (MNU)-induced multi-organ carcinogenesis in F344 rats

Acrylamide (AA) has been reported to be formed in fried and baked foods with various concentrations, and exposure levels to AA from cooked foods in children are estimated to be higher than those in adults. In order to evaluate the carcinogenicity of AA exposure during childhood, we conducted a medium-term carcinogenicity study with prepubertal administration of AA followed by treatments of a multi-organ-targeted genotoxic carcinogen and a promoting agent for thyroid carcinogenesis in rats. A total of 36 postpartum F344 rats were given drinking water containing AA at 0, 20, 40 or 80 ppm for 3 weeks during the lactation period, and their weaned offspring received the same AA-containing water for 3 more weeks. Offspring were then injected with N-methyl-N-nitrosourea (MNU; 40 mg/kg body weight, i.p.) once at week 7 after birth. Half the animals of the 0 and 40 ppm groups were additionally treated with the anti-thyroid agent sulfadimethoxine (SDM; 125 ppm) in the drinking water thereafter. Offspring were subjected to complete necropsy at week 50. All the major organs and macroscopic abnormalities were excised and examined histopathologically. There was no significant difference in the incidences of hyperplastic and neoplastic lesions in the target organs of AA and/or MNU, such as the brain, spinal cord, pituitary gland, thyroid, adrenal glands, uterus, mammary glands, clitoral gland and tunica vaginalis. In conclusion, no significant modifying actions of AA on MNU-induced multi-organ carcinogenesis were exhibited in any organs of rats when exposed prepubertally under the present experimental conditions.

Study on Influence of Apitherapy in Acrylamide Experimentally Induced Liver Disease Regarding the Hepatic Enzyme Profile in Wistar Rats

Introduction. The purpose of this experiment was to study the influence of apitherapy in acrylamide-induced liver disease in rats. The experimental study objectives were the evaluation of liver enzyme parameters (ALT, TGO, AP, GGT), in conditions of chronic liver damage induced by acrylamide. To reduce the factors that accelerate the progress of liver damage we have administered Apitherapy products. Materials and methods. Materials: Apitherapy products Apiregya, ApiImunomod, ApiImunostim. The Apitherapy products were purchased from Stupina LLC. The animals were handled under thiopental induced general anesthesia. Determining the level of investigated parameters, i.e. serum total protein was measured using an automatic analyzer (Aeroset, Abbott) and commercial kits (Abbott, USA). A total of 40 Wistar white rats were used in the study; they were divided evenly into four groups: standard diet control group (group I), apitherapy and royal jelly control group (group II), acrylamide group (group III), acrylamide-apitherapy- royal jelly control group (group IV). The animals were kept under standard light and temperature conditions with access to food and water ad libitum. The toxic liver disease in rats was experimentally induced by gastric gavage administration of acrylamide (in aqueous solution, 50 mg /kg body weight). Results and discussions. Administration of apitherapy and royal jelly (RJ) products in experimental animals with acrylamide–induced hepatopathy (Group IV) produced a reduction towards normal limits of liver enzymes compared to the other studied groups, as follows: i). compared to the control group with standard diet (Group I), for ALT (93.85  2.26 vs. 110.66  10.52); AST (64.78  1.47 vs. 57.94  9.1), alkaline phosphatase (112.42  4.99 vs. 86.44  6.93), GGT (0.571  0.07 vs. 0.233  0.11); ii) compared to the group treated with apitherapy and RJ (Group IV), for: ALT (55.28  1.49 vs. 110.66  10.52); AST (35.35  0.71 vs. 57.96  9.1), alkaline phosphatase (97.28  1.38 vs. 86.44  6.93), GGT (0.457  0.05 vs. 0.233  0.11). The administration of the apitherapy + RJ to the group which received the first the acrylamide (group IV) was effective for maintaining the TGO values within close limits to the standard food group values (group II); bringing within the absolute normal limits for TGP value and bringing within limits comparable with the values of group II, which received the apitherapy and royal jelly. Conclusions. It is recommended to administer the Apitherapy preparations Apiregya, ApiImunostim, ApiImunomod, for normalizing the liver enzymes in liver damage induced by toxic food. Acknowledgments. This paper was financially supported by the European Social Fund and the Romanian Government under project PERFORM-ERA (ID-57649).

Study of the Influence Of Apitherapy in Acrylamide Experimentally Induced Liver Disease Regarding Blood Count Parameters in Wistar Rats

Introduction. The purpose of this experiment was to study the influence of apitherapy in acrylamide-induced liver disease in rats. The experimental study objectives were the evaluation of blood count parameters in conditions of chronic liver damage induced by acrylamide. To reduce the factors that accelerate the progress of liver damage we have administered Apitherapy products. Materials and Methods. Materials: Apitherapy products Apiregya, ApiImunomod, ApiImunostim. The Apitherapy products were purchased from Stupina LLC. The animals were handled under thiopental induced general anesthesia. Determining the level of investigated parameters, i.e. serum total protein was measured using an automatic analyzer (Aeroset, Abbott) and commercial kits (Abbott, USA). A total of 40 Wistar white rats were used in the study; they were divided evenly into four groups: standard diet control group (group I), apitherapy and royal jelly control group (group II), acrylamide group (group III), acrylamide-apitherapy- royal jelly control group (group IV). The animals were kept under standard light and temperature conditions with access to food and water ad libitum. The toxic liver disease in rats was experimentally induced by gastric gavage administration of acrylamide (in aqueous solution, 50 mg /kg body weight). Results and discussions. The administrationof the apitherapy + RJ to the group that first received the acrylamide (group IV) was effective for maintaining the parameters within limits that were close to the standard food values group (group II) regarding the leukocyte count, erythrocyte number, hemoglobin, hematocrit, VEM, HEM, CHEM, erythrocyte distribution width, thrombocyte count, average thrombocyte volume, width of thrombotic distribution. Conclusions. The study proves that Apitherapy preparations like Apiregya, ApiImunostim, ApiImunomod are efficient in treating liver damage disease and they normalize the liver blood count. Acknowledgments: Financing was done by Human Resources Development Operational Programme through performance by integrating doctorate in European research area project (PERFORM-ERA) ID 57649.

Study On The Influence Of Apitherapy In Acrylamide Experimentally Induced Liver Disease Regarding The Protein Profile In Wistar Rats

Introduction. The purpose of this experiment was to study the influence of the apitherapy in acrylamide-induced liver disease in rats. The experimental study objectives were the evaluation of serum protein electrophoresis parameters (albumin, globulin, alpha-1- globulin, alpha-2 globulin, beta globulin, albumin ratio/globulin), in conditions of chronic liver damage induced by acrylamide. To reduce the factors that accelerate progress of liver damage we have administered Apitherapy products. Materials and methods. Materials: Apiregya Apitherapy products, ApiImunomod, ApiImunostim. The apitherapy products were purchased from Stupina LLC”. The animals were handled under thiopental induced general anesthesia. Determining the level of investigated parameters, i.e. serum total protein was measured using an automatic analyzer (Aeroset, Abbott) and commercial kits (Abbott, USA). A total of 40 Wistar white rats were used in the study; they were divided evenly into four groups: standard diet control group (group I), apitherapy and royal jelly control group (group II), acrylamide group (group III), acrylamide-apitherapy- royal jelly control group (group IV). The animals were kept under standard light and temperature conditions with access to food and water ad libitum. The toxic liver disease in rats was experimentally induced by gastric gavage administration of acrylamide (in aqueous solution, 50 mg /kg body weight). Results and discussions. In comparison to the other treated groups, administration of apidiet and royal jelly (RJ) in experimental animals with acrilamide–induced hepatopathy (Group IV) produced: i) an increase in total proteins compared to the Control Group [Group I] (6.150.03 vs. 7.330.01) and a reduced value for this parameter compared to the Group II (7.330.01 vs. 6.280.33), ii) an increased level of albumin compared to the Control Group [Group I] (30.140.69 vs. 41.71.34), Group II (40.710.95 vs. 41.71.34), iii) an increase of albumin/globulin ratio compared to Group I (0.4320.01 vs. 0.7200.04), and a maintainig of this ration compared to Group II (0.60.03 vs. 0.7200.04), iiii) a reduction of levels of globulins compared to the Control Group [Group I] (69.850.69 vs. 58.411.12), with similar results for globulins as those obtained for Group II (59.141.06 vs. 58.411.12). Regarding the globulins fractions, we can remarks these are returned to normal values in the animals with acrilamide–induced hepatopathy and treated with apidiet and royal jelly. Conclusions. It is recommended to administer apitherapy preparations Apiregya, ApiImunostim, ApiImunomod for normalizing the parameters of serum protein electrophoresis in liver damage induced by toxic food. Acknowledgments. Financing was done by Human Resources Development Operational Programme (PERFORM-ERA) ID 57649.

Acrylamide induces cell death in neural progenitor cells and impairs hippocampal neurogenesis

Acrylamide (ACR) is a well-known neurotoxin in mammalian species that causes neuropathy characterized by ataxia and skeletal muscle weakness. Therefore, ACR-mediated axon damage in the central and peripheral nervous systems is considered to be central-peripheral axonopathy. However, the molecular mechanisms underlying ACR's toxicity to neural progenitor cells are unknown. This study investigated the adverse effects of ACR on mouse multipotent neural progenitor cells and adult hippocampal neurogenesis. ACR significantly reduced the proliferation of neural progenitor cells, and high ACR concentrations induced apoptotic and necrotic cell death. We found that elevated intracellular levels of reactive oxygen species were involved in ACR-mediated cytotoxicity. Interestingly, the administration of ACR to young mice resulted in a significant decrease in the number of newly generated cells in the dentate gyrus of the hippocampus, suggesting an impairment of adult neurogenesis. These results suggest that ACR's deleterious effects on the central nervous system are due to the death of neural progenitor cells and impaired adult neurogenesis.

Increased H-ras mutation frequency in mammary tumors of rats initiated with N-methyl-N-nitrosourea (MNU) and treated with acrylamide

We recently demonstrated the incidence and multiplicity of N-methyl-N-nitrosourea (MNU)-induced mammary tumors to be increased by administration of acrylamide (AA) in post-initiation in rats. In the present study, to clarify the mechanisms of enhancement, H-ras gene mutations in mammary tumors induced in MNU-initiated rats with or without subsequent AA administration were investigated. Frequencies of mutations in codon 12 from GGA to GAA were significantly (p < 0.05) higher in rats with AA administration (82%, 23 out of 28 tumors) as compared to those without AA (50%, 9 out of 18 tumors), but the latency and volume of H-ras mutation-harboring tumors were similar to those of the mutation-lacking tumors. No mutations in codons 13 or 61 were detected in either treatment groups. The results thus indicate that H-ras gene mutations in codon 12 play a pivotal role in initiation of carcinogenesis and it appears possible that AA administration may selectively co-stimulate and/or maintain initiated cells via other genomic or non-genomic events in MNU-treated rats.

Effect of Dose Volume on the Toxicokinetics of Acrylamide and Its Metabolites and 2-Deoxy-d-glucose

Acrylamide (AA) is a known mutagen and animal carcinogen. Comparison of recent studies revealed significant quantitative differences in AA-induced germ cell mutagenicity. It was hypothesized that despite the administration of AA at similar doses, the discrepancy in the observed effects was most likely due to varying AA concentrations in the administered dosing solution. To test this hypothesis, AA was administered i.p. to mice at 50 mg/kg in a dose volume of 5 or 50 ml/kg, blood was collected at various time points, and AA and its metabolites were quantitated. Changes in dose volume resulted in significant differences in the toxicokinetics of AA and its metabolites and suggested that increased C(max) of AA led to increased metabolism. This theory, in conjunction with the fact that higher levels of AA-derived radioactivity were detected in the testes, may explain the greater toxicity of a 50 mg/kg dose when administered in 5 versus 50 ml/kg. The impact of dose volume on the toxicokinetics of 2-deoxy-d-glucose (DG), a nonreactive, nonmetabolizable substance, was also investigated. The areas under the curve for DG were not different for the two dose volumes; however, C(max) for the more concentrated dose was significantly higher. In conclusion, current studies show that the toxicokinetics of an administered xenobiotic and its metabolites is influenced by the concentration of the parent chemical in the dosing solution. Therefore, it is important to consider the concentration of an administered xenobiotic in the dosing solution because it may affect its toxicokinetics and metabolism and subsequently affect the biological effects of the administered chemical.

Acrylamide does not induce tumorigenesis or major defects in mice in vivo

Chronic administration of acrylamide has been shown to induce thyroid tumors in rat. In vitro acrylamide also causes DNA damage, as demonstrated by the comet assay, in various types of cells including human thyroid cells and lymphocytes, as well as rat thyroid cell lines. In this work, mice were administered acrylamide in their drinking water in doses comparable with those used in rats, i.e., around 3-4 mg/kg per day for mice treated 2, 6, and 8 months. Some of the mice were also treated with thyroxine (T(4)) to depress the activity of the thyroid. Others were treated with methimazole that inhibits thyroid hormone synthesis and consequently secretion and thus induces TSH secretion and thyroid activation. These moderate treatments were shown to have their known effect on the thyroid (e.g. thyroid hormone and thyrotropin serum levels, thyroid gland morphology...). Besides, T(4) induced an important polydipsia and degenerative hypertrophy of adrenal medulla. Acrylamide exerted various discrete effects and at high doses caused peripheral neuropathy, as demonstrated by hind-leg paralysis. However, it did not induce thyroid tumorigenesis. These results show that the thyroid tumorigenic effects of acrylamide are not observed in another rodent species, the mouse, and suggest the necessity of an epidemiological study in human to conclude on a public health policy.

Alteration of Oxidative Status in Rats Following Administration of Acrylamide

Introduction: Acrylamide (ACR) is a known industrial neurotoxic and carcinogenic chemical in rodents. The recent discovery of acrylamide in wide variety of commonly consumed foods has energized research efforts worldwide to define toxic mechanisms. Objective: The present study is carried out to investigate the effect of acrylamide administration on in vivo malondialdehyde (MDA, a product of lipid peroxidation), reduced glutathione (GSH) as well as copper and zinc superoxide dismutase enzyme activity (Cu/Zn SOD) of rats. Material and Methods: Fourteen adult male Sprague Dawley rats were divided into two groups each containing 7 rats. Group 1 served as negative control fed on basal diet and group 2 (positive control) received basal diet and acrylamide (0.34g/ kg diet) for 11 days. Levels of MDA, GSH and activity of SOD were determined in liver, kidneys, brain, heart, testes, spleen and lungs of rats. Results: ACR treatment significantly increased MDA in all organs; the highest increase was detected in testis (87.9%) and heart (71.5%) while the lowest one was found in kidneys (28.2%). On the other hand, GSH levels and SOD activities were significantly reduced in ACR treated rats. However, the reduction of GSH level ranged from 10.2% to 36.5 %.The inhibition of SOD activities were higher in testis (57.3%) and lungs (38.5%). Conclusion: The present study showed that ACR exerts deteriorated effects on oxidative

Alteration of Oxidative Status in Rats Following Administration of Acrylamide

Introduction: Acrylamide (ACR) is a known industrial neurotoxic and carcinogenic chemical in rodents. The recent discovery of acrylamide in wide variety of commonly consumed foods has energized research efforts worldwide to define toxic mechanisms. Objective: The present study is carried out to investigate the effect of acrylamide administration on in vivo malondialdehyde (MDA, a product of lipid peroxidation), reduced glutathione (GSH) as well as copper and zinc superoxide dismutase enzyme activity (Cu/Zn SOD) of rats. Material and Methods: Fourteen adult male Sprague Dawley rats were divided into two groups each containing "7" rats. Group 1 served as negative control fed on basal diet and group 2 (positive control) received basal diet and acrylamide (0.34g/ kg diet) for 11 days. Levels of MDA, GSH and activity of SOD were determined in liver, kidneys, brain, heart, testes, spleen and lungs of rats. Results: ACR treatment significantly increased MDA in all organs; the highest increase was detected in testis (87.9%) and heart (71.5%) while the lowest one was found in kidneys (28.2%). On the other hand, GSH levels and SOD activities were significantly reduced in ACR treated rats. However, the reduction of GSH level ranged from 10.2% to 36.5 %.The inhibition of SOD activities were higher in testis (57.3%) and lungs (38.5%). Conclusion: The present study showed that ACR exerts deteriorated effects on oxidative

Excretion of mercapturic acids of acrylamide and glycidamide in human urine after single oral administration of deuterium-labelled acrylamide

We investigated the human metabolism of AA to the mercapturic acids N-acetyl-S-(2-carbamoylethyl)-L-cysteine (AAMA) and N-(R/S)-acetyl-S-(2-carbamoyl-2-hydroxyethyl)-L: -cysteine (GAMA) which are derived from AA itself and from its oxidative genotoxic metabolite glycidamide (GA), respectively. A healthy male volunteer received a single dose of about 1 mg deuterium-labelled acrylamide (d(3)-AA), representing 13 microg/kg body weight, in drinking water. Urine samples before dosing and within 46 h after the dose were analysed for d(3)-AAMA and d(3)-GAMA by LC-ESI-MS/MS. A first phase of increase in urinary concentration was found to last 18 h with a broad plateau between 8 and 18 h for AAMA, and 22 h for GAMA. Elimination half-lives of both AAMA and GAMA were estimated to be approximately 3.5 h for the first phase and more than 10 h up to few days for the second phase. Total recovery in urine after 24 h was about 51% as the sum of AAMA and GAMA and hereby well in accordance with former studies in rats. After 2 days AAMA, accounting for altogether 52% of the total AA dose, was the major metabolite of AA in humans. GAMA, accounting for 5%, appeared as a minor metabolite of AA. In humans we found a urinary ratio of 0.1 for GAMA/AAMA compared to previously reported values of 0.2 for rats and 0.5 for mice. Therefore, the metabolic fate of AA in humans was more similar to that in rats than in mice as already demonstrated in terms of the haemoglobin adducts. Consequently a genotoxic potency of AA mediated by GA could be supposed to be comparable in rats and humans.

Role of CYP2E1 in the Epoxidation of Acrylamide to Glycidamide and Formation of DNA and Hemoglobin Adducts

Acrylamide (AA) is an animal carcinogen, neurotoxin, and reproductive toxin. AA is formed in baked and fried carbohydrate-rich foods. Metabolism of AA occurs via epoxidation to glycidamide (GA) or direct conjugation with glutathione. Using CYP2E1-null mice, recent studies in this laboratory demonstrated that induction of somatic and germ cell mutagenicity in AA-treated mice is dependent on CYP2E1. We hypothesized that AA metabolism to GA is a prerequisite for the induction of AA-induced mutagenicity. Current studies were undertaken to assess the role of CYP2E1 in the epoxidation of AA to GA and the formation of DNA and hemoglobin (HGB) adducts. AA was administered to CYP2E1-null or wild-type mice at 50 mg/kg ip. Mice were euthanized 6 h later and blood and tissues were collected. Using LC-ES/MS/MS, AA, GA, and DNA- and HGB-adducts were measured. While the plasma levels of AA and GA were 115 +/- 14.0 and 1.7 +/- 0.31 microM in CYP2E1-null mice, they were 0.84 +/- 0.80 and 33.0 +/- 6.3 microM in the plasma of AA-treated wild-type mice. Administration of AA to wild-type mice caused a large increase in N7-GA-Gua and N3-GA-Ade adducts in the liver, lung, and testes. While traces of N7-GA-Gua adducts were measured in the tissues of AA-treated CYP2E1-null mice, these levels were 52- to 66-fold lower than in wild-type mice. Significant elevation of both AA- and GA-HGB adducts was detected in AA-treated wild-type mice. In AA-treated CYP2E1-null mice, levels of AA-HGB adducts were roughly twice as high as those in wild-type mice. In conclusion, current work demonstrated that CYP2E1 is the primary enzyme responsible for the epoxidation of AA to GA, which leads to the formation of GA-DNA and HGB adducts.

DNA damage and DNA adduct formation in rat tissues following oral administration of acrylamide

Acrylamide is present as a contaminant in the human diet in heated food products. It has been found to be carcinogenic in laboratory rats and has been classified as probably carcinogenic in humans. In order to clarify the possible involvement of a primary genotoxic mechanism in acrylamide-induced carcinogenicity, both the presence of DNA damage, measured by the comet assay, and the formation of N7-(2-carbamoyl-2-hydroxyethyl)guanine (N7-GA-Gua) and N3-(2-carbamoyl-2-hydroxyethyl)adenine (N3-GA-Ade), derived from reaction of the active metabolite glycidamide (GA) with the DNA, analyzed by LC/MS/MS, were assessed in selected rat tissues. Rats were administered with single oral doses of acrylamide (18, 36 or 54 mg/kg body weight (b.w.) and the organs (blood leukocytes, brain, bone marrow, liver, testes and adrenals) were sampled at different times after treatment. Results from GA-induced DNA adduct measurements indicated a relatively even organ distribution of the adducts in brain, testes and liver. Organ-specificity in acrylamide carcinogenesis can therefore not be explained by a selective accumulation of GA–DNA adducts in the target organs, at least not after a single dose exposure. The DNA adduct profiles and half-lives were similar in the different organs; except that the N3-GA-Ade adduct was more rapidly removed from tissues than the N7-GA-Gua adduct. Increased extent of DNA migration, as measured by the in vivo rat comet assay, was found in brain and testes, and these specific results seem to be in accordance with the known organ-specificity in acrylamide carcinogenesis in rat. Only weak and transient DNA damage was recorded in the liver, bone marrow and adrenals. The DNA-damaging effect of the compound observed in the blood leukocytes could be a simple biomarker of acrylamide exposure and genotoxicity.