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1.
Exp Toxicol Pathol ; 69(7): 533-546, 2017 Sep 05.
Article in English | MEDLINE | ID: mdl-28502747

ABSTRACT

Chicken egg fetal livers were evaluated for histopathological changes produced by four genotoxic hepatocarcinogens: 2-acetylaminofluorene (AAF), aflatoxin B1 (AFB1), benzo[a]pyrene (BaP), diethylnitrosamine (DEN); four structurally related non- or weakly- carcinogenic comparators: fluorene (FLU), aflatoxin B2 (AFB2), benzo[e]pyrene (BeP), N-nitrosodiethanolamine (NDELA); two epigenetic hepatocarcinogens: clofibric acid (CFA), phenobarbital (PB); and the non-carcinogen, D-mannitol (MAN). CFA, PB and MAN were also assessed for formation of DNA adducts using the 32P nucleotide postlabeling (NPL) assay and for DNA breaks using the comet assay. CFA was also assessed in enhanced comet assay for oxidative DNA damage induction. Eggs were dosed on days 9- 11 of incubation. For genotoxicity evaluation, livers were collected 3h after the last dose. Liver qualitative histopathology assessment was performed on days 12 and 18 of incubation. CFA was negative for DNA adducts but yielded clear evidence of DNA breaks due to oxidative stress. PB and MAN produced no DNA adducts or breaks. Liver to body weight ratios were not affected in most groups, but were decreased in DEN groups, and increased after PB dosing. Livers from control groups, FLU, AFB2, BeP, NDELA, CFA, and MAN groups, displayed a typical hepatocellular trabecular pattern at both time points. In contrast, the four genotoxic carcinogens induced time- and dose- related interference with fetal liver cell processes of proliferation, migration and differentiation, leading to hepatocellular and cholangiocellular pleomorphic dysplasia and re-(de-) differentiation with distortion of the trabecular pattern. In addition, dosing with the high dose of DEN produced gallbladder agenesis. PB induced hepatocellular hypertrophy, interference with migration, expressed as distortion of the trabecular pattern, and a moderate cholangiocellular dysplasia. In summary, histopathological analysis of chicken fetal livers revealed developmental anomalies, as well as genotoxicity-induced and, in the case of PB, adaptive morphological changes. Thus, the model provides histopathological outcomes of molecular effects.


Subject(s)
Carcinogens/toxicity , Liver/drug effects , Mutagenicity Tests/methods , Animals , Chick Embryo , Comet Assay , DNA/analysis , DNA/genetics
2.
Exp Toxicol Pathol ; 67(9): 453-8, 2015 Sep.
Article in English | MEDLINE | ID: mdl-26164753

ABSTRACT

Propoxur (PPX) is a carbamate insecticide which induced urinary bladder cancer in Wistar rats when fed at 5000ppm in Altromin 1321 diet (1321). In the present investigation, PPX was studied for induction of several key events related to modes of action (MOA) of carcinogenicity in urinary bladders (UBs). Wistar rats were administered the compound for 28 days at 8000ppm in Provini Liba SA 3883 diet, which is similar to the 1321 diet. o-Anisidine HCl (AH) was used as a genotoxic UB carcinogenic comparator, and trisodium nitrilotriacetate (NTA) as an epigenetic UB carcinogen comparator. Along with the non-dosed control and three test substance groups (PPX, AH, NTA), four more groups were additionally fed 2% ammonium chloride (AC) in the diet to acidify the urine, since 1321 was reported to increase urinary pH. AC did acidify the urine, as expected, although the 3883 diet itself did not increase pH values above 8. In the alkaline comet assay, AH produced DNA single strand breaks (SSBs) in the UB urothelium (UBU) irrespective of AC administration, whereas PPX and NTA did not. In the nucleotide (32)P-postlabeling assay (NPL), AH produced DNA adducts irrespective of AC administration, whereas PPX and NTA did not. Routine (H&E) histopathology evaluation of the UBU did not reveal any hyperplasia or evidence of luminal microprecipitates or calculi in any of the groups. Assessment of UBU proliferation as measured by immunohistochemistry of proliferating cell nuclear antigen, revealed that NTA and NTA plus AC increased the replicating fraction (RF). Also AH plus AC, but not AH alone, increased the RF of UBU, whereas PPX groups were not significantly different from controls. Thus, the results reveal no evidence for DNA SSBs, binding, or alteration of DNA synthesis in the UBU by PPX, while demonstrating UBU DNA damage by AH and showing that NTA does not damage DNA, but causes increased UBU proliferation. The findings are in accord with a genotoxic MOA for AH, and an epigenetic MOA for NTA. The MOA of PPX does not involve genotoxicity and may be specific to the 1321 diet.


Subject(s)
DNA Adducts/metabolism , DNA Damage , Insecticides/toxicity , Mutagens/toxicity , Propoxur/toxicity , Urinary Bladder/drug effects , Administration, Oral , Animals , Cell Proliferation/drug effects , Comet Assay , Male , Rats, Wistar , Urinary Bladder/metabolism , Urinary Bladder/pathology , Urothelium/drug effects , Urothelium/metabolism , Urothelium/pathology
3.
Exp Toxicol Pathol ; 65(4): 451-6, 2013 May.
Article in English | MEDLINE | ID: mdl-22459205

ABSTRACT

Human liver cancer is in part associated with obesity and related metabolic diseases. The present study was undertaken in a mouse model of diet-induced obesity (DIO) and hepatic steatosis, conditions which can be associated with hepatic neoplasia, to determine whether the rates of cell proliferation or hepatocarcinogen bioactivation were altered in ways which could facilitate hepatocarcinogenesis. DIO mice were generated by feeding C57BL/6 (B6) male mice a high-fat diet beginning at 4 weeks of age; age-matched conventional lean (LEAN) B6 mice fed a low fat diet (10% Kcal from fat) were used for comparison. Groups of 28 week old DIO and LEAN mice were dosed with the bioactivation-dependent DNA-reactive hepatocarcinogen 2-acetylaminofluorene (AAF), at 2.24 or 22.4 mg/kg, given by gavage 3 times per week for 31 days, or received no treatment (DIO and LEAN control groups). Compared with the LEAN control group, the DIO control group had a higher mean body weight (16.5 g), higher mean absolute (1.4 g) and mean relative (25.5%) liver weights, higher (394%) liver triglyceride concentrations, and an increased incidence and severity of hepatocellular steatosis at the end of the dosing phase. The DIO control group also had a higher mean hepatocellular replicating fraction (31% increase, determined by proliferating cell nuclear antigen immunohistochemistry). Hepatocarcinogen bioactivation, based on formation of AAF DNA adducts as measured by nucleotide (32)P-postlabeling, was similar in both DIO and LEAN AAF-dosed groups. Thus, hepatocellular proliferation, but not hepatocarcinogen bioactivation, was identified as an alteration in livers of DIO mice which could contribute to their susceptibility to hepatocarcinogenesis.


Subject(s)
Cell Proliferation/drug effects , Fatty Liver/physiopathology , Hepatocytes/drug effects , Obesity/complications , 2-Acetylaminofluorene/analogs & derivatives , 2-Acetylaminofluorene/toxicity , Animal Feed , Animals , Carcinogens/toxicity , DNA Adducts/analysis , DNA Adducts/biosynthesis , Diet, Fat-Restricted , Diet, High-Fat/adverse effects , Disease Models, Animal , Mice , Mice, Inbred C57BL , Obesity/physiopathology
4.
Regul Toxicol Pharmacol ; 53(2): 107-20, 2009 Mar.
Article in English | MEDLINE | ID: mdl-19027814

ABSTRACT

In response to a Hazard Notice by the Medical Devices Agency of the UK in 2000 regarding the Trilucent breast implant (TBI), an expert panel was convened to implement a research program to determine whether genotoxic compounds were formed in the soybean oil filler (SOF) of TBIs and whether these could be released to produce local or systemic genotoxicity. The panel established a research program involving six laboratories. The program recruited 47 patients who had received TBIs (9 patients had received silicone implants previously). A reference group (REBI) of 34 patients who had exchanged either silicone (17 patients) implants (REBI-E) or patients (17) who were to receive primary implantation augmentation with silicone (REBI-PIA), and who were included as needed to increase either the pre- or post-explantation sample number. Of the 17 REBI-E patients, 5 had silicone implants and 12 had saline implants previously (prior to the last exchange). Investigation was undertaken before and after replacement surgery in the TBI patients and before and after replacement or augmentation surgery in the REBI patients. The pre- to post-operative sample interval was 8-12 weeks. Pre-operative samples were collected within 7 days prior to the operation. Information on a variety of demographic and behavioral features was collected. Biochemical and biological endpoints relating to genotoxic lipid peroxidation (LPO) products potentially formed in the SOF, and released locally or distributed systemically, were measured. The SOF of explanted TBIs was found to have substantial levels of LPO products, particularly malondialdehyde (MDA), and low levels of trans-4-hydroxy-2-nonenal (HNE) not found in unused implants. Mutagenicity of the SOF was related to the levels of MDA. Capsules that formed around TBIs were microscopically similar to those of reference implants, but MDA-DNA adducts were observed in capsular macrophages and fibroblasts of only TBI capsules. These cell types are not progenitors of breast carcinoma (BCa) and the location of the implants precludes LPO products reaching the mammary epithelial cells which are progenitors of BCa. Blood levels of LPO products were not increased in TBI patients compared to REBI patients and did not change with explantation. In TBI patients, white blood cells did not show evidence of increased levels of LPO-related aldehyde DNA adducts. In conclusion, based on a number of measured parameters, there was no evident effect that would contribute to breast or systemic cancer risk in the TBI patients, and the recommended treatment of TBI patients involving explantation was judged appropriate.


Subject(s)
Breast Implants/adverse effects , Lipid Peroxidation , Mutagenicity Tests , Soybean Oil/adverse effects , Adult , Aldehydes/metabolism , Device Removal , Female , Fibroblasts/metabolism , Humans , Macrophages/metabolism , Malondialdehyde/metabolism , Middle Aged , Prosthesis Failure , Silicone Gels , Sodium Chloride/chemistry
5.
Exp Toxicol Pathol ; 57(5-6): 397-404, 2006 Jul.
Article in English | MEDLINE | ID: mdl-16616835

ABSTRACT

The chronic toxicity and carcinogenicity of Nifurtimox (NFX), a 5-nitrofuran derivative used in the treatment of American trypanosomiasis, were studied in male and female Wistar rats in an accelerated cancer bioassay (ACB). The ACB is a mechanistic initiation/promotion chronic toxicity and carcinogenicity bioassay designed to assess potential carcinogenic activity of a test substance in critical organs and tissues of rodents in which human carcinogens are active. The organs studied were liver, lungs, urinary bladder (UB), mammary gland (MG), bone marrow, spleen, kidneys, colon, stomach and any grossly observed lesions. NFX is a genotoxin which has been reported previously to exert a variable degree of carcinogenic activity in rat liver, kidney, UB and MG. The present study was undertaken to assess whether NFX has initiating activity in these four named target sites. In the initiation phase, groups of 20 Wistar rats were given NFX daily in the diet at 0.2% for the first 12 weeks of the study to assess initiating activity, followed by promoters (PROs) for four organs for an additional 24 weeks. NFX was compared to the following known initiators (INs) for each of these four tissues: diethylnitrosamine (DEN) for liver and kidney, N-butyl-N(4-hydroxybutyl)nitrosamine (BBN) for UB and 7,12-dimethylbenz(a)anthracene (DMBA) for MG. PROs included phenobarbital (PB) for liver and kidney, nitrilotriacetic acid (NTA) for UB, and diethylstilbestrol (DES) for MG. NFX was also administered continuously without PROs for 40 weeks. At the end of dosing (40 weeks) and at the end of recovery (52 weeks), animals were sacrificed and subjected to complete gross and histopathological examinations, along with evaluations of body weight gain over time and terminal body weights. Mortality was highest with DEN+PB (group 6) (40%), followed by BBN+NTA (group 7) (15%) and NFX+DES (group 5) and DMBA+DES (group 8) (10% each). The same groups also showed significant reductions in body weight gain over time and terminal body weights at sacrifice. In these groups, the expected preneoplastic, neoplastic and metastatic neoplastic lesions were produced, demonstrating the sensitivity of the model. In groups given NFX+PROs (groups 3-5), either no neoplasms occurred (group 4) or only single neoplasms (groups 3 and 5). In contrast, the PROs all elicited tumors in groups given INs (groups 6-8). Also, NFX given alone for 40 weeks did not produce any chronic toxicity, preneoplastic or neoplastic lesions. Thus, in this study, NFX did not demonstrate chronic toxicity or carcinogenicity. Moreover, in four target sites, i.e., liver, kidney, UB and MG, it exhibited no neoplastic initiating activity manifested by PROs for these four target sites.


Subject(s)
Antiparasitic Agents/toxicity , Neoplasms, Experimental/etiology , Nifurtimox/toxicity , Animals , Antiparasitic Agents/classification , Body Weight/drug effects , Carcinogenicity Tests , Cocarcinogenesis , Drug Therapy, Combination , Female , Longevity/drug effects , Male , Nifurtimox/classification , Rats , Rats, Wistar , Toxicity Tests, Chronic , Weight Gain/drug effects
6.
Eur J Cancer Prev ; 11(1): 39-48, 2002 Feb.
Article in English | MEDLINE | ID: mdl-11917207

ABSTRACT

A previous investigation demonstrated the anticarcinogenicity of acetaminophen (APAP) against colon carcinogenesis in rats induced by 3,2'-dimethyl-4-aminobiphenyl (DMAB). DMAB was selected as a structurally related surrogate for heterocyclic amines, formed during cooking of protein, which are believed to be involved in human colon cancer. The objective of the present study was to ascertain whether the early initiating effects of this colon carcinogen are inhibited by APAP. Six groups of male F344 rats were treated over a 6-week period as follows: (1) vehicle (corn oil) for 6 weeks; (2) APAP in the diet at 1000 ppm daily for 6 weeks; (3) 50 mg/kg DMAB by gavage once a week for the last 4 weeks; (4) 5 mg/kg DMAB as for (3); (5) 1000 ppm APAP for 6 weeks and 50 mg/kg DMAB for the last 4 weeks; and (6) 1000 ppm APAP and 5 mg/kg DMAB as for (5). Colonic tissue was within normal limits in the control and APAP groups. In the APAP only group, apical enterocytic hypertrophy and hyperaemia over the entire surface epithelium was present. In the high-dose DMAB group, in the lower third of the crypts, foci of enlarged glands with hypertrophic cells exhibiting karyomegaly and anisokaryosis (FHE) of 3+ degree of severity were evident in 100% of the animals. Also, there were increases in periglandular fibrocytes, matrix and mononuclear cells (PF). In the low-dose DMAB group both FHE and PF changes with the same degree of severity were reduced. In rats given the low dose of DMAB plus APAP, FHE and PF with the same degree of severity (3+) was absent. Both DMAB exposures increased significantly the replicating fraction of colonic enterocytes in an exposure-related fashion and the replicating fractions were significantly reduced by APAP. In 32P-postlabelling of colon, liver and urinary bladder DNA, high-dose DMAB produced 2-6 distinct dose-related spots reflecting DNA adducts. These spots were reduced or were no longer detectable in all three tissues when APAP was given 2 weeks before and during DMAB exposure. Using immunohistochemical detection of DMAB adducts in the colon, a dose-related colour intensity was present for both doses of DMAB. APAP reduced this by 94-fold. Thus, APAP produced a marked protective effect in colonic enterocytes against several parameters of neoplastic development by the carcinogen.


Subject(s)
Acetaminophen/pharmacology , Aminobiphenyl Compounds/antagonists & inhibitors , Anticarcinogenic Agents/pharmacology , Carcinogens/antagonists & inhibitors , Colonic Neoplasms/prevention & control , Acetaminophen/administration & dosage , Animals , Anticarcinogenic Agents/administration & dosage , Colonic Neoplasms/chemically induced , Colonic Neoplasms/chemistry , DNA Adducts/analysis , Dose-Response Relationship, Drug , Immunohistochemistry , Male , Rats , Rats, Inbred F344 , Severity of Illness Index , Time Factors
7.
Eur J Cancer Prev ; 11 Suppl 2: S101-7, 2002 Aug.
Article in English | MEDLINE | ID: mdl-12570342

ABSTRACT

The synthetic monocyclic phenolics (MPs), acetaminophen (APAP), butylated hydroxyanisole (BHA), and butylated hydroxytoluene (BHT) are antimutagenic or anticarcinogenic against a diversity of chemical carcinogens affecting a variety of tissues in experimental animals. In studies in this laboratory of the anticarcinogenicity of MPs, the focus has been on delineating efficacy at low levels of MPs that do not elicit adaptive or toxic responses. To accomplish this, we are studying anticarcinogenicity against the neoplastic initiating activity of lower doses of carcinogens than have previously been studied and which are closer to human environmental exposures. In these studies, we have investigated anticarcinogenicity of BHT against liver cancer in rats induced by either 2-acetylaminofluorene (AAF) or aflatoxin B1 (AFB1) and anticarcinogenicity of APAP against colon cancer induced in rats by 3,2'-dimethyl-4-aminobiphenyl (DMAB). BHA and BHT at 100-125 ppm in the diet inhibited the initiation phase of AAF and AFB1 hepatocarcinogenesis and therefore may act intracellularly to block effects of the carcinogen. Likewise, with APAP in colon anticarcinogenicity, at 1000 ppm it reduced DNA binding and exerted a cytoprotective effect against DMAB. Thus, APAP also shows evidence of producing a blocking effect. We conclude that these MPs appear to be anticarcinogenic through a mechanism different from that of most other chemopreventive agents, possibly involving interception of the reactive chemical species of the carcinogen. Accordingly, they have promise as cancer prophylactics, including in combination with agents operating through other mechanisms.


Subject(s)
Acetaminophen/pharmacology , Anticarcinogenic Agents/pharmacology , Butylated Hydroxyanisole/pharmacology , Butylated Hydroxytoluene/pharmacology , Chemoprevention/methods , Neoplasms/prevention & control , Animals , Anticarcinogenic Agents/administration & dosage , Humans , Rats , Sensitivity and Specificity
8.
Arch Toxicol ; 75(1): 52-8, 2001 Mar.
Article in English | MEDLINE | ID: mdl-11357521

ABSTRACT

Alternative bioassays of mannitol (MAN) and caprolactam (CAP) were conducted in transgenic p53-deficient mice. Also, to assess the sensitivity of the transgenic mice to a model DNA-reactive carcinogen, the hepatic effects of diethylnitrosamine (DEN) were compared in the wild type background strain of mouse and in the transgenic derivative. Fifty-one male wild type strain C57BL/6 mice p53 (+/+), 8 weeks old, and 51 heterozygous p53 (+/-) C57BL/6 Tac-[KO] Trp53 N5 mice, 8 weeks old, were allocated to six experimental groups as follows: groups 1 (wild type +/+) and 2 (p53 +/-) served as room controls, groups 3 (+/+) and 4 (+/-) were exposed orally (gavage) to 50 mumol/kg body weight DEN weekly for a total of ten doses during the first 10 weeks of the study, group 5 (+/-) was exposed to 15,000 ppm CAP in the diet for up to 26 weeks, and group 6 (+/-) was exposed to 50,000 ppm MAN in the diet for up to 26 weeks. After 10 weeks, liver from control and DEN-exposed mice was used for O4-ethylthymidine (O4-EtT) DNA adduct analysis by the immunoslot blot method. The cell replicating fraction (RF) in the liver was determined by quantification of the percentage of immunohistochemically stained hepatocytes positive for proliferating cell nuclear antigen. No significant or consistent body or liver weight changes were present in any of the treatment groups. No consistent and pertinent changes in RF values were present in any of the treatment groups. None of the tested substances produced neoplasms of any type in p53 (+/-) mice. DEN induced comparable levels of O4-EtT adducts in the liver in both wild type and p53 +/- genotypes, but no morphologic changes were evident in the livers of either genotype. The lack of response to DEN, in spite of formation of DNA adducts, may reflect the resistance to hepatocarcinogenesis of the background C57BL/6 strain of the transgenic, and calls into question the general sensitivity of this transgenic for detection of carcinogenic effects.


Subject(s)
Caprolactam/toxicity , Diethylnitrosamine/toxicity , Genes, p53/physiology , Liver/drug effects , Mannitol/toxicity , Administration, Oral , Alkylating Agents/toxicity , Animals , Apoptosis/drug effects , Biological Assay , Body Weight/drug effects , Caprolactam/administration & dosage , Carcinogens/toxicity , Cell Division/drug effects , DNA Adducts/drug effects , Diethylnitrosamine/administration & dosage , Dose-Response Relationship, Drug , Heterozygote , Immunohistochemistry , Liver/metabolism , Liver/pathology , Mannitol/administration & dosage , Mice , Mice, Inbred C57BL , Mice, Transgenic , Proliferating Cell Nuclear Antigen/metabolism
9.
Exp Toxicol Pathol ; 53(5): 345-57, 2001 Oct.
Article in English | MEDLINE | ID: mdl-11817103

ABSTRACT

The chronic toxicity and carcinogenicity of Moxifloxacin (MOX), a bacterial gyrase-inhibiting fluoroquinolone antibiotic, were studied in male and female Wistar rats in an accelerated cancer bioassay (ACB). The ACB is a mechanistic initiation/promotion chronic toxicity and carcinogenicity study designed to assess potential carcinogenic activity of a test substance in critical organs in which human carcinogens are active. The organs studied were liver, lungs, urinary bladder, mammary gland, bone marrow, thymus, spleen and stomach. MOX was given daily by intragastric instillation at 500 mg/kg bw/day for the first 13 weeks to produce potential initiation, followed by promoters (PROs) for 24 weeks, or for the last 24 weeks after 13 weeks of exposure to initiators (INs). The INs, administered during the first 13 weeks, were diethylnitrosamine for the liver, N-n-butyl-N-(4-hydroxybutyl)nitrosamine for the urinary bladder, ethylnitrosourea for the hematolymphoreticular system, N-nitrosodimethylamine for lungs, methylnitrosourea for the stomach and 7,12-dimethylbenz(a)-anthracene for the mammary gland. The PROs, administered during the last 24 weeks after MOX, were phenobarbital for the liver, nitrilotriacetic acid for the urinary bladder, azathioprine for the bone marrow, butylated hydroxytoluene for the lung, butylated hydroxyanisole for the forestomach, and diethylstilbestrol for the mammary gland. The INs produced preneoplastic and neoplastic lesions which were not enhanced by MOX, and MOX plus PROs elicited no neoplastic effects, documenting that MOX did not produce either initiation or promotion of neoplasia in any of the target sites, or in any of the other twenty tissues examined.


Subject(s)
Anti-Infective Agents/toxicity , Aza Compounds , Carcinogens/toxicity , Fluoroquinolones , Neoplasms, Experimental/chemically induced , Quinolines , Administration, Oral , Animals , Body Weight/drug effects , Carcinogenicity Tests/methods , Cocarcinogenesis , Female , Male , Moxifloxacin , Organ Size/drug effects , Rats , Rats, Wistar , Time Factors
10.
Toxicol Pathol ; 28(3): 388-95, 2000.
Article in English | MEDLINE | ID: mdl-10862555

ABSTRACT

To explore differences in mechanisms of carcinogenicity at low and high exposures, we have conducted a series of exposure-response studies of hepatocarcinogenesis in rats using 2 well-studied DNA-reactive carcinogens, 2-acetylaminofluorene and diethylnitrosamine. In these studies, we have used intraperitoneal injection or intragastric instillation to deliver exact doses during an initiation segment followed by phenobarbital as a liver tumor promoter to enhance manifestation of initiation. This protocol results in carcinogenicity comparable to that produced by lifetime exposure to the carcinogens. Our findings in these experiments provide evidence for the following: (a) formation of DNA adducts can be nonlinear, with a plateau at higher exposures; (b) cytotoxicity shows no-effect levels and is related to exposure; (c) compensatory hepatocyte proliferation shows no-effect levels and can be supralinear at high exposures; (d) formation of preneoplastic hepatocellular altered foci can show no-effect levels and appears supralinear at high exposures; (e) no-effect levels can exist for tumor development, and the exposure response can be supralinear. We interpret these findings to reflect thresholds for hepatocellular initiating effects of these carcinogens and exaggerated responses at high exposures attributable to cytotoxicity and compensatory hepatocyte proliferation. Such enhanced proliferation of hepatocytes harboring DNA damage likely results in an exaggerated yield of mutations in critical genes, leading to supralinear initiation of carcinogenesis. Thus, mechanisms differ between low and high exposures. Based on these observations, we suggest that linear extrapolation from high toxic exposures to postulated low-exposure effects of DNA-reactive carcinogens can yield overestimates. Such extrapolation must be supported by mechanistic information. The finding of no-effect levels provides a basis for understanding why low-level environmental exposures of humans to even DNA-reactive carcinogens may convey no cancer risk.


Subject(s)
2-Acetylaminofluorene/toxicity , Carcinogens/toxicity , DNA Adducts/drug effects , Diethylnitrosamine/toxicity , Liver Neoplasms, Experimental/chemically induced , Liver/drug effects , Animals , DNA Damage/drug effects , Dose-Response Relationship, Drug , Liver/metabolism , Liver/pathology , Liver Neoplasms, Experimental/metabolism , Liver Neoplasms, Experimental/pathology , Male , Nonlinear Dynamics , Rats , Rats, Inbred F344
12.
Arch Toxicol ; 73(7): 394-402, 1999 Sep.
Article in English | MEDLINE | ID: mdl-10550482

ABSTRACT

In previous exposure-response studies, we have documented non-linearities for some of the early effects in rat liver of diethylnitrosamine (DEN) and a near no-effect levels for initiation of promotable liver neoplasms at the lowest cumulative exposure of 0. 5 mmol/kg body weight; this in spite of formation of DNA adducts and induction of hepatocellular altered foci (HAF). To extend these investigations, in an initiation segment, young male F344 rats were administered four exposures of DEN ranging from a cumulative total of 0.25 mmol, which is half of the previously used low exposure, up to 2 mmol per kg body weight, an effective initiating exposure. These exposures were achieved by once weekly intragastric instillations of one-tenth the total exposures for up to 10 weeks. The initiation segment was followed by a 4 week recovery segment, to allow for remission of acute and subchronic effects of DEN, after which the groups were maintained on 0.06% phenobarbital in the diet for 24 weeks to promote liver tumor development in order to assess initiation. During and after initiation and at the end of recovery, selected groups were studied for several crucial effects involved in hepatocarcinogenicity. The low exposure produced a low-level of DNA ethylation at both 5 and 10 weeks of exposure, measured as O(4)-ethylthymidine, the most persistent promutagenic ethylation product. At the 5 week interval, the adduct values of the higher exposures were less than proportional to the increment of exposure, suggestive of nonlinearity. Assessment of cellular proliferation by staining for proliferating cell nuclear antigen revealed that the lowest exposure did not increase the replicating fraction of hepatocytes during the initiation (10 weeks) or recovery (4 weeks) segments, whereas in the three higher exposure groups, proliferation was increased in relation to dose and time. Preneoplastic HAF expressing glutathione S-transferase-placental-type were present at low multiplicity in control livers and their multiplicity was increased in all exposure groups by the end of exposure, at which time the increase in the high exposure group was disproportionately greater than the increment of exposure. After phenobarbital administration in the promotion segment, all exposure groups exhibited further HAF increases at 39 weeks. At the end of the promotion segment, no hepatocellular neoplasm was found in 80 controls or in 40 rats in the low exposure group. In the mid-low exposure group, which was the previously studied low exposure, only one adenoma was found, yielding a 3% incidence, while in the two higher exposure groups, 32 and 80% of rats exhibited liver neoplasms, which were increased disproportionately greater than the increments of exposure. Thus, the findings document non-linearities of early DEN effects and at the lowest cumulative dose, a no-effect level (NEL) or threshold for initiation of promotable liver neoplasms. These findings provide a conceptual basis for understanding why low-level exposures to DNA-reactive carcinogens may convey no cancer risk.


Subject(s)
Carcinogens/toxicity , DNA Adducts/drug effects , Diethylnitrosamine/toxicity , Liver Neoplasms, Experimental/chemically induced , Liver/drug effects , Animals , Carcinogens/administration & dosage , Cell Division/drug effects , DNA Damage/drug effects , Diethylnitrosamine/administration & dosage , Dose-Response Relationship, Drug , Ethylenes , Glutathione S-Transferase pi , Glutathione Transferase/metabolism , Isoenzymes/metabolism , Linear Models , Liver/metabolism , Liver/pathology , Liver Neoplasms, Experimental/metabolism , Liver Neoplasms, Experimental/pathology , Male , No-Observed-Adverse-Effect Level , Proliferating Cell Nuclear Antigen/metabolism , Rats , Rats, Inbred F344
13.
Food Chem Toxicol ; 37(9-10): 1027-38, 1999.
Article in English | MEDLINE | ID: mdl-10541460

ABSTRACT

Butylated hydroxyanisole (BHA) and butylated hydroxytoluene (BHT) are widely used antioxidant food additives. They have been extensively studied for potential toxicities. This review details experimental studies of genotoxicity and carcinogenicity which bear on cancer hazard assessment of exposure to humans. We conclude that BHA and BHT pose no cancer hazard and, to the contrary, may be anticarcinogenic at current levels of food additive use.


Subject(s)
Antioxidants/toxicity , Butylated Hydroxyanisole/toxicity , Butylated Hydroxytoluene/toxicity , Food Additives , Administration, Oral , Animals , Anticarcinogenic Agents/metabolism , Butylated Hydroxyanisole/administration & dosage , Butylated Hydroxyanisole/metabolism , Butylated Hydroxytoluene/administration & dosage , Butylated Hydroxytoluene/metabolism , Carcinogenicity Tests , Carcinogens , Consumer Product Safety , Diet , Dose-Response Relationship, Drug , Methylnitronitrosoguanidine , Mutagenicity Tests
14.
Toxicol Pathol ; 26(6): 759-68, 1998.
Article in English | MEDLINE | ID: mdl-9864093

ABSTRACT

Uterine Cell proliferation was studied in intact Sprague-Dawley (SD) and Fischer 344 (F344) rats exposed to the antiestrogens tamoxifen (TAM; 5, 10, 20, or 40 mg/kg) and toremifene (TOR: 21.2 or 42.4 mg/kg). The antiestrogens were administered to animals via gavage daily for 2 or 12 wk. Uterine proliferation was assessed using markers for the proliferating cell nuclear antigen (PCNA) and by the bromodeoxyuridine (BrdU) method. Diethylstilbestrol (DES) was used as an estrogenic reference compound. The antiestrogens either reduced or prevented changes of myometrial and stromal proliferation indices (PI). TAM and TOR caused a time-dependent reduction of endometrial glands without an associated decrease in cell proliferation. In the luminal columnar epithelium, the antiestrogens depressed PCNA PI but enhanced BrdU PI, indicating a low continuous DNA synthesis in otherwise quiescent cells. The antiestrogens induced focal hyperplastic multilayered epithelia with PCNA-positive basal cells along segments of the luminal uterine epithelium. We suggest that this hyperplastic epithelium represents remnants from the glandular epithelium. DES was less efficient in inducing these changes but induced squamous metaplasias in the F344 rats. Uterine effects of the 2 antiestrogens were comparable with the exception of I TAM-exposed (40 mg/kg) SD rat that showed squamous metaplasia. F344 rats were more sensitive to the estrogenic action of DES than were the SD rats.


Subject(s)
Estrogen Antagonists/toxicity , Tamoxifen/toxicity , Toremifene/toxicity , Uterus/drug effects , Animals , Atrophy/chemically induced , Atrophy/pathology , Body Weight/drug effects , Bromodeoxyuridine/analysis , Cell Division/drug effects , Diethylstilbestrol/toxicity , Endometrium/drug effects , Endometrium/pathology , Epithelial Cells/drug effects , Epithelial Cells/pathology , Female , Myometrium/drug effects , Myometrium/pathology , Proliferating Cell Nuclear Antigen/analysis , Rats , Rats, Inbred F344 , Rats, Sprague-Dawley , Species Specificity , Stromal Cells/drug effects , Stromal Cells/pathology , Uterus/chemistry , Uterus/pathology
15.
Toxicol Sci ; 45(2): 152-61, 1998 Oct.
Article in English | MEDLINE | ID: mdl-9848122

ABSTRACT

The dose responses for several effects of low-level limited exposures to 2-acetylaminofluorene (AAF) in the livers of male Fischer 344 rats were measured and a subsequent phenobarbital tumor promotion regimen was used to manifest initiation of carcinogenesis. Three doses over a 10-fold range yielding cumulative total exposures of 0.126, 0.42, and 1.26 mmol AAF/kg body weight were achieved by daily intragastric instillation for up to 12 weeks with interim terminations. This was followed by 24 weeks administration of 500 ppm phenobarbital (PB) in the diet to promote liver tumor development. At 12 weeks at the end of AAF administration, all exposures produced adducts in liver DNA, measured by 32P postlabeling, and the level of adducts increased with exposure, except that the high exposure did not produce a dose proportional increase. Measurement of arylsulfotransferase activity, a key enzyme in the metabolic activation of AAF, revealed that in livers from the high exposure animals, the enzyme was inhibited. To assess for toxicity, the centrilobular zone of glutamine synthetase-positive hepatocytes was quantified immunohistochemically at 12 weeks. The area of the zone was reduced in the high exposure group and there was a trend to reduction in relationship to exposure. The two lower exposures to AAF produced no increase in cell proliferation, whereas the high exposure resulted in a marked increase, about 8-fold over controls. Initiation was assessed by induction of hepatocellular altered foci (HAF) that expressed the placental form of glutathione S-transferase. AAF induced HAF in the high exposure group, 9-fold at 8 weeks and 170-fold at 12 weeks compared to controls. In rats maintained on PB for 24 weeks after exposure, the multiplicity of HAF increased in controls and comparably in the low and mid exposure groups, but remained at the about the same high level in the high exposure group. The high exposure produced a substantial incidence of benign neoplasms by 12 weeks, and with promotion by 36 weeks, all rats developed hepatocellular neoplasia. In the mid exposure group, only one adenoma occurred at 36 weeks in 17 rats, while in the low exposure group, no liver tumor occurred in 23 rats. Thus, these findings document nonlinearities for some of the effects of AAF, with supralinear effects at the high exposure for cell proliferation and induction of HAF, and a no-observed-effect level for induction of promotable liver neoplasms at the lowest cumulative exposure of 0.126 mmol/kg, in spite of the formation of DNA adducts. We conclude that the effects of this DNA-reactive hepatocarcinogen leading to initiation exhibit nonlinearities and possible thresholds.


Subject(s)
2-Acetylaminofluorene/toxicity , Carcinogens/toxicity , Liver Neoplasms, Experimental/chemically induced , Liver/drug effects , Animals , Arylsulfotransferase/metabolism , Cell Division/drug effects , DNA Adducts/metabolism , Dose-Response Relationship, Drug , Liver/metabolism , Liver/pathology , Male , Rats , Rats, Inbred F344
16.
Cancer Res ; 58(18): 4096-101, 1998 Sep 15.
Article in English | MEDLINE | ID: mdl-9751618

ABSTRACT

Here, we examined the effect of black tea and caffeine on lung tumorigenesis in F344 rats induced by the nicotine-derived carcinogen 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) in a 2-year bioassay. NNK was administered s.c. at a dose of 1.5 mg/kg body weight three times weekly for 20 weeks. Animals were given either black tea as drinking water at concentrations of 2%, 1%, or 0.5%, or caffeine in drinking water at concentrations identical to those in 2% and 0.5% tea infusions for 22 weeks. The treatment period began 1 week before and ended 1 week after the NNK administration. The animals were sacrificed on week 101 for the examination of tumors in target organs, including lung, liver, nasal cavity, and other major organs. The NNK-treated group, given 2% black tea, showed a significant reduction of the total lung tumor (adenomas, adenocarcinomas, and adenosquamous carcinomas) incidence from 47% to 19%, whereas the group given 1% and 0.5% black tea showed no change. The 2% tea also reduced liver tumor incidence induced by NNK from 34% in the group given only deionized water to 12%. The tumor incidence in the nasal cavity, however, was not affected by either black tea or caffeine at any of the concentrations tested. The most unexpected finding was the remarkable reduction of the lung tumor incidence, from 47% to 10%, in the group treated with 680 ppm caffeine, a concentration equivalent to that found in the 2% tea. This incidence is comparable to background levels seen in the control group. This study demonstrated for the first time in a 2-year lifetime bioassay that black tea protects against lung tumorigenesis in F344 rats, and this effect appears to be attributed, to a significant extent, to caffeine as an active ingredient of tea.


Subject(s)
Anticarcinogenic Agents/pharmacology , Caffeine/pharmacology , Lung Neoplasms/prevention & control , Tea/chemistry , Animals , Body Weight/drug effects , Carcinogens , Drug Screening Assays, Antitumor , Liver Neoplasms, Experimental/chemically induced , Lung Neoplasms/chemically induced , Male , Neoplasms, Experimental/chemically induced , Neoplasms, Experimental/prevention & control , Nitrosamines , Nose Neoplasms/chemically induced , Rats , Rats, Inbred F344 , Survival Rate
17.
Carcinogenesis ; 18(11): 2247-53, 1997 Nov.
Article in English | MEDLINE | ID: mdl-9395228

ABSTRACT

A striking difference between two structurally related anti-estrogen medicines is that tamoxifen is strongly hepatocarcinogenic in the rat, whereas toremifene lacks such activity. To study the basis for this difference, the initiating potential of tamoxifen and toremifene were studied by measurement of rapid induction of hepatocellular altered foci (HAF) that express placental-type glutathione S-transferase in the livers of female Sprague-Dawley (S-D) rats and female Fischer 344 (F344) rats. Both agents were administered by gavage at equimolar doses up to a dose that produced marked weight gain suppression. In rats given the high dose of 40 mg/kg per day tamoxifen continuously for 36 weeks, 75% of S-D rats developed liver neoplasms, in contrast to only 10% of F344 rats. In the S-D strain, tamoxifen produced a tendency to increased HAF at 2 weeks at the dose of 40 mg/kg per day and by 12 weeks, a dose-related increase was evident. In contrast, toremifene induced no HAF even at the equimolar high dose of 42.4 mg/kg per day for 12 weeks. The induction of HAF by tamoxifen was less in the F344 rats. Neither agent elicited increases in hepatocellular proliferation in S-D or F344 rats. When phenobarbital was administered for 24 weeks as a promoting agent after the anti-estrogens, S-D rats given tamoxifen at 20 mg/kg per day for 12 weeks, developed liver neoplasms, but not F344 rats or rats of either strain given even a higher dose (42.4 mg/kg) of toremifene. Thus, tamoxifen has initiating activity in these rat strains whereas toremifene does not.


Subject(s)
Estrogen Antagonists/toxicity , Liver Neoplasms, Experimental/chemically induced , Precancerous Conditions/chemically induced , Tamoxifen/toxicity , Toremifene/toxicity , Animals , Body Weight/drug effects , Female , Liver/drug effects , Liver/pathology , Rats , Rats, Inbred F344 , Rats, Sprague-Dawley
18.
Eur J Cancer Prev ; 6(4): 357-62, 1997 Aug.
Article in English | MEDLINE | ID: mdl-9370098

ABSTRACT

The widely used analgesic acetaminophen (APAP) was studied in rats for its ability to inhibit intestinal carcinogenesis induced by 3,2'-dimethyl-4-aminobiphenyl (DMAB), which was selected as the carcinogen because of its similarity to the heterocyclic amines formed during cooking and which are postulated to be involved in colon cancer in humans. APAP was fed to male F344 rats at 250 ppm, which is about 1/4 the human therapeutic dose and at 5000 ppm, which is about fivefold the human dose. DMAB was injected subcutaneously at 50 mg/kg body weight weekly for 20 weeks, to assure identical exposures to all animals, followed by 22 weeks of maintenance. The DMAB was an effective inducer of tumours in the small and large intestines, producing an average of 1.3 tumours per animal. Feeding of APAP began 2 weeks before DMAB administration and continued for 44 weeks. A 9% reduction in the number of colon tumours per rat cancer at the low dose and an 86% reduction at the high dose were found. Small intestinal tumour incidence was reduced at both doses. The number of multiple intestinal tumours per rat was reduced by 27% and 49% for the low and high doses, respectively. The dimensions of these neoplasms, especially those in the colon, were also reduced in both dose groups. Thus, APAP, even at a sub-therapeutic dose, inhibited intestinal carcinogenesis induced by DMAB. This allows us to speculate that the effects of low exposures to dietary carcinogens of the heterocyclic amine type could be inhibited by therapeutic doses of APAP.


Subject(s)
Acetaminophen/administration & dosage , Analgesics, Non-Narcotic/administration & dosage , Carcinogens/antagonists & inhibitors , Intestinal Neoplasms/drug therapy , Mutagens/pharmacology , Neoplasms, Experimental/drug therapy , Aminobiphenyl Compounds , Animals , Colorectal Neoplasms/drug therapy , Colorectal Neoplasms/mortality , Colorectal Neoplasms/prevention & control , Disease Models, Animal , Dose-Response Relationship, Drug , Food Contamination , Intestinal Neoplasms/chemically induced , Intestinal Neoplasms/mortality , Intestine, Large/drug effects , Intestine, Large/pathology , Intestine, Small/drug effects , Intestine, Small/pathology , Male , Neoplasms, Experimental/chemically induced , Rats , Rats, Inbred F344 , Survival Rate
19.
Exp Toxicol Pathol ; 49(3-4): 153-65, 1997 Aug.
Article in English | MEDLINE | ID: mdl-9314049

ABSTRACT

The chronic toxicity and carcinogenicity of Wingstay 100 (W 100), a rubber antioxidant/antiozonant, were studied in Fischer 344 (F 344) rats in two chronic studies. Earlier genetic studies indicated that the product had weak activity in a bacterial mutation assay, but lacked activity in chromosomal aberration assays. In an one year study, both genders of F 344 rats were exposed to 53, 310 or 1900 ppm in NIH-07 diet for 52 weeks, and sacrifices were made at 38, 52 and 64 weeks. No test substance related deaths occurred, although the high dose of 1900 ppm caused a decrease in body weight gain and food consumption in both genders. Red blood cell mean corpuscular volume was significantly increased at 38 weeks, accompanied by a significant decrease in mean corpuscular hemoglobin concentration. At 52 weeks, the red blood cell count and hemoglobin values were also significantly decreased in high dose animals of both genders. Total bilirubin and cholesterol were increased in high dose animals of 38 and 52-week sacrifices. During the 3 month recovery, hematology parameters, bilirubin and cholesterol returned to control values. Total protein was reduced in high dose animals of both genders, throughout the entire exposure and recovery periods. W 100 also produced increases in relative liver, spleen, heart and kidney weights in high dose animals. Both genders of all W 100 groups exhibited significant increases in urothelial cell proliferation (measured by PCNA) and adaptive hyperplasia. No regenerative hyperplasia, preneoplasia, or neoplasia were present. There was microscopic evidence of extramedullary erythropoiesis in the spleen and liver of high dose animals in both genders, otherwise no other pertinent microscopic finding was evident. In parallel, an accelerated bioassay (ABA) was conducted, which is a mechanistic initiation/promotion carcinogenicity study designed to assess tumor induction and promotion potential of a test substance in major organs of carcinogenesis. The present study was conducted in male F 344 rats for 38 weeks. The target sites chosen for the ABA were liver and urinary bladder and the dose for W 100 was 1900 ppm previously established to be a toxic dose. The liver tumor initiator was diethylnitrosamine (DEN), and the urinary bladder initiator was N-butyl N-(4-hydroxybutyl) nitrosamine (BBN). The initiators were administered during the first 14 weeks followed by the promoters. The promoters, phenobarbital (PB) for the liver and nitrilotriacetate (NTA) for the urinary bladder, were administered during the last 24 weeks of the study after the test substance. The study had 11 test groups including a negative control. The critical comparisons for initiating activity were conducted between groups 3 (PB) and 6 (W 100 + PB) for the liver and groups 8 (NTA) and 11 (W 100 + NTA) for the urinary bladder. The critical comparisons for promoting activity were conducted between groups 2 (DEN) and 5 (DEN + W 100) for the liver and groups 7 (BBN) and 10 (BBN + W 100) for the urinary bladder. There were 26 and 38-week sacrifices. In this study, most body weight reductions were due to DEN. At 26 weeks, significant increases in liver weights were present in all PB-exposed groups. Significant increases in renal weights occurred in all NTA, BBN and DEN groups. A similar organ weight pattern was present at 38 weeks. At 26 weeks, there were hepatocellular (33%) and urothelial (67%) tumors present in positive control groups (DEN/DEN + PB/BBN/BBN + NTA). In contrast, in the DEN + W 100 (5) and the BBN + W 100 (10) groups no tumors were present indicating absence of promotion. In addition, no tumors were present in groups 6 (W 100 + PB) or 11 (W 100 + NTA) indicating absence of initiation. At 38 weeks, the incidences of hepatocellular adenomas and carcinomas in positive control group (DEN) was 44%. The incidence of urothelial adenomas and carcinomas was 67% in group 7 (BBN). In contrast, groups 5 (DEN + W 100) or group 10 (BBN + W 100) had


Subject(s)
Antioxidants/toxicity , Carcinogens/toxicity , Phenylenediamines/toxicity , Animals , Bilirubin/blood , Cholesterol/blood , Erythrocyte Indices/drug effects , Female , Kidney/pathology , Kidney Neoplasms/chemically induced , Kidney Neoplasms/pathology , Liver/pathology , Liver Neoplasms, Experimental/chemically induced , Liver Neoplasms, Experimental/pathology , Male , Myocardium/pathology , Neoplasms, Experimental/chemically induced , Organ Size/drug effects , Phenylenediamines/administration & dosage , Rats , Rats, Inbred F344 , Sex Characteristics , Spleen/pathology , Weight Gain/drug effects
20.
Exp Toxicol Pathol ; 49(1-2): 15-28, 1997 Feb.
Article in English | MEDLINE | ID: mdl-9085070

ABSTRACT

Salicylazosulfapyridine (SASP), which has been in clinical use for over 50 years, was reported by the National Toxicology Program to increase rat (F344 strain) urinary bladder and mouse (B6C3F1 hybrid) liver tumours under ad libitum (AL) feeding conditions, while under a feed restriction (FR) regimen, these tumours were not increased. The present investigations were undertaken to assess the implications of these results for the safety of SASP in humans. SASP and its 2 major metabolites, 5-aminosalicylic acid (ASA) and sulfapyridine (SP) were tested for in vivo induction of micronuclei in mouse bone marrow cells with or without prefolic treatment and for in vivo formation of DNA adducts in rat and mouse liver and urinary bladder. None exhibited mutagenicity or DNA reactivity. SASP and SP have induced sister chromatid exchanges and micronuclei (MN) in cultured human lymphocytes in the absence of liver activation enzymes and in B6C3F1 mice (but not in rats) MN in bone marrow and peripheral RBC. Treatment with folate reduces the frequency of MN. Perhaps the short (28 days) RBC lifespan in mouse underlies the sensitivity of this species. Thus, SASP without folate supplementation is an aneuploidogen. In a 2-year study in AL fed SASP-treated (high dose 337.5 mg/kg) rats, urinary pH was increased and urinary specific gravity was reduced at 60 weeks. At the end, this SASP group showed urothelial hyperplasia and papillomas in the urinary bladders of male rats primarily. In the FR high dose SASP group, the hyperplasia was reduced from 82% to 14%. At the end of 2 years, the incidence of multi-organ leukemia was reduced in both AL and FR high dose SASP groups. Thus, SASP caused intraluminal bladder changes in the rat (especially males) consisting of chronic urothelial stimulation, concretions, hyperplasia which resulted in neoplasia. In the mouse, because of species differences in liver ratios (mouse > rat) and, increasing (3 times higher) liver perfusion in the mouse, compared to the rat, there was hepatocellular toxicity and resulting preneoplasia and neoplasia within 2 years. These findings occurred in all AL SASP groups (flat curve without dose response); but were reduced under FR conditions. In this species, the multiorgan lymphoma incidence was reduced in both AL and FR high dose SASP groups. Thus, SASP and its major metabolites are not genotoxic. Folate deficiency associated with SASP administration is probably responsible for aneuploidy in lymphocytes and erythrocytes. SASP does not induce neoplasia directly in either livers, urinary bladders or other organs. Accordingly, SASP is judged to pose no carcinogenic risk to humans.


Subject(s)
Anti-Inflammatory Agents/toxicity , Bone Marrow/drug effects , Liver/drug effects , Sulfasalazine/toxicity , Urinary Bladder/drug effects , Aminosalicylic Acids/pharmacokinetics , Aminosalicylic Acids/toxicity , Animals , Anti-Infective Agents/pharmacokinetics , Anti-Infective Agents/toxicity , Anti-Inflammatory Agents/pharmacokinetics , Carcinogenicity Tests , DNA Adducts/drug effects , Female , Folic Acid/pharmacology , Male , Mesalamine , Mice , Micronuclei, Chromosome-Defective/drug effects , Mutagenicity Tests , Rats , Rats, Sprague-Dawley , Risk Assessment , Sulfapyridine/pharmacokinetics , Sulfapyridine/toxicity , Sulfasalazine/pharmacokinetics
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