RESUMEN
8-Hydroxydeoxyguanosine (8-OHdG) is well known not only as an effective biomarker of oxidative stress but also as a mutagenic DNA modification. Incorporation of dAMP at the opposite site of 8-OHdG induces G>T or A>C transversions. However, in vivo analyses of gene mutations caused by potassium bromate (KBrO3), which can induce 8-OHdG at carcinogenic target sites, showed that G>T was prominent in the small intestines of mice, but not in the kidneys of rats. Because KBrO3 was a much clearer carcinogen in the kidneys of rats, detailed analyses of gene mutations in the kidney DNA of rats treated with KBrO3 could improve our understanding of oxidative stress-mediated carcinogenesis. In the current study, site-specific reporter gene mutation assays were performed in the kidneys of gpt delta rats treated with KBrO3. Groups of 5 gpt delta rats were treated with KBrO3 at concentrations of 0, 125, 250, or 500 ppm in the drinking water for 9 weeks. At necropsy, the kidneys were macroscopically divided into the cortex and medulla. 8-OHdG levels in DNA extracted from the cortex were dramatically elevated at concentrations of 250 ppm and higher compared with those from the medulla. Cortex-specific increases in mutant frequencies in gpt and red/gam genes were found at 500 ppm. Mutation spectrum and sequence analyses of their mutants demonstrated significant elevations in A>T transversions in the gpt gene and single base deletions at guanine or adenine in the gpt or red/gam genes. While A>T transversions and single base deletions of adenine may result from the oxidized modification of adenine, the contribution of 8-OHdG to gene mutations was limited despite possible participation of the 8-OHdG repair process in guanine deletion.
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Bromatos , ADN , Riñón , Ratas , Ratones , Animales , 8-Hidroxi-2'-Desoxicoguanosina , Mutación , Adenina , Carcinogénesis , Carcinógenos , GuaninaRESUMEN
Rubiadin (Rub) is a genotoxic component of madder color (MC) that is extracted from the root of Rubia tinctorum L. MC induces renal tumors and preneoplastic lesions that are found in the proximal tubule of the outer stripe of the outer medulla (OSOM), suggesting that the renal carcinogenicity of MC is site specific. To clarify the involvement of Rub in renal carcinogenesis of MC, we examined the distribution of Rub in the kidney of male gpt delta rats that were treated with Rub for 28 days. We used desorption electrospray ionization quadrupole time-of-flight mass spectrometry imaging (DESI-Q-TOF-MSI), along with the histopathological analysis, immunohistochemical staining, and reporter gene mutation assays of the kidney. DESI-Q-TOF-MSI revealed that Rub and its metabolites, lucidin and Rub-sulfation, were specifically distributed in the OSOM. Histopathologically, karyomegaly characterized by enlarged nuclear and microvesicular vacuolar degeneration occurred in proximal tubule epithelial cells in the OSOM. The ɤ-H2AX- and p21-positive cells were also found in the OSOM rather than the cortex. Although dose-dependent increases in gpt and Spi- mutant frequencies were observed in both the medulla and cortex, the mutant frequencies in the medulla were significantly higher. The mutation spectra of gpt mutants showed that A:T-T:A transversion was predominant in Rub-induced gene mutations, consistent with those of MC. Overall, the data showed that the distribution of Rub and its metabolites resulted in site-specific histopathological changes, DNA damage, and gene mutations, suggesting that the distribution of genotoxic components and metabolites is responsible for the site-specific renal carcinogenesis of MC.
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Daño del ADN , Riñón , Ratas , Masculino , Animales , Ratas Endogámicas F344 , Riñón/patología , CarcinogénesisRESUMEN
Elemicin, an alkenylbenzene flavoring, exists naturally in foods, herbs, and spices. Some alkenylbenzenes are hepatotoxic and hepatocarcinogenic in rodents. However, few studies have examined the toxicology of elemicin. In the current study, we comprehensively evaluated the general toxicity, genotoxicity, and carcinogenicity of elemicin using gpt delta rats and DNA adductome analysis. Groups of 10 male F344 gpt delta rats were treated with elemicin by gavage at a dose of 0, 25, 100, or 400 mg/kg bw/day for 13 weeks. Liver weights were significantly increased with histopathological changes in groups receiving 100 mg/kg bw/day or more. Significant increases in serum hepatotoxic parameters were observed in the 400 mg/kg bw/day group. Based on the observed changes in liver weights, 18.6 mg/kg bw was identified as the low benchmark dose. Significant increases in the number and area of glutathione S-transferase placental form-positive foci and gpt mutant frequencies were apparent only in the 400 mg/kg/day group, although elemicin-specific DNA adducts were detected from the lowest dose, suggesting that elemicin exhibited hepatocarcinogenicity in rats only at higher doses. Because elemicin showed no mutagenicity at lower doses, there was an adequate safety margin between the acceptable daily intake and the estimated daily intake of elemicin.
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Aromatizantes , Placenta , Embarazo , Ratas , Masculino , Femenino , Animales , Ratas Endogámicas F344 , Pruebas de MutagenicidadRESUMEN
2-Methylfuran (2-MF) exists naturally in foods and is used as a flavoring agent. Furan, the core structure of 2-MF, possesses hepatocarcinogenicity in rodents. Accumulation of toxicological information on furan derivatives is needed to elucidate their carcinogenic mode of action. In the current study, we examined the comprehensive toxicological studies of 2-MF using gpt delta rats. 2-MF was intragastrically administered to groups of 10 male and 10 female Sprague-Dawley gpt delta rats at a dose of 0, 1.2, 6, or 30 mg/kg/day for 13 weeks. Effects of 2-MF on the hepatobiliary system including an increase in serum alkaline phosphatase were observed in the 6 and 30 mg/kg groups, and cholangiofibrosis was found in the 30 mg/kg group. The no observed adverse effect level was set at 1.2 mg/kg/day for both sexes and 1.14 mg/kg/day was determined as the benchmark dose low. The acceptable daily intake was calculated to be 11.4 µg/kg/day. Increases in the number and areas of glutathione S-transferase placental form-positive foci in the 30 mg/kg group were apparent, suggesting the hepatocarcinogenicity of 2-MF in rats. By contrast, the lack of increase in in vivo mutagenicity in the liver implied that 2-MF hepatocarcinogenesis may not involve genotoxic mechanisms.
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Fosfatasa Alcalina , Aromatizantes , Animales , Carcinógenos/toxicidad , Daño del ADN , Relación Dosis-Respuesta a Droga , Femenino , Aromatizantes/farmacología , Furanos/toxicidad , Glutatión Transferasa , Hígado , Masculino , Pruebas de Mutagenicidad , Placenta , Embarazo , Ratas , Ratas Endogámicas F344 , Ratas Sprague-Dawley , Ratas TransgénicasRESUMEN
Benchmark dose (BMD) method have been used in the toxicological assessment of chemical substances so that the point of departure can be derived, as an alternative to the use of no observable adverse effect level (NOAEL), and the method is often applied to the incidence data of histopathological findings in the toxicity studies. In the present study, the BMD method was applied to various patterns of incidence data derived from some toxicity studies as case studies, and the validity of each application was discussed. Five independent applications including toxicity studies of madder color or semicarbazide hydrochloride were prepared and model averaging over the three models with the lowest three AIC (Akaike information criteria) values (MA-3), a recently proposed model averaging method, was employed. The series of case studies indicated, for the better application of the BMD method to histopathological findings, the following points:(i) If there are incidence data with severity grading of pathologically significant lesions, we must discuss whether the BMD method should be applied to the total incidence data or the incidence data above certain grade with or without data aggregation.(ii) If a lesion of interest had higher toxicological significance rather than the secondary lesions with higher severity, the BMD method should be applied to the incidence data of the lesion of interest.(iii) If it is highly necessary to apply the BMD method to obtained incidence data without toxicological and statistical validity, toxicological pathologists are advised to review individual datasets of histopathology and associated data, and provide new incidence data of comprehensive findings (diagnostic name) such as hepatocellular injury or nephropathy, if possible. In all cases, toxicological significance and mechanism of a lesion of interest need to be considered in light of the dose-dependence. In view of both toxicology and statistics, sufficient discussions must be made on the validity of applying BMD method and its estimate.
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Benchmarking , Relación Dosis-Respuesta a Droga , Incidencia , Nivel sin Efectos Adversos Observados , Medición de RiesgoRESUMEN
There has been no report on the prevalence of Campylobacter spp. in farm animals in Mongolia. To uncover the prevalence of Campylobacter spp. in chickens in Mongolia and their antimicrobial resistance, in this study, we isolated and characterized Campylobacter spp. from chickens in Mongolia. We collected 71 cloacal swabs of chickens from 5 farms including 4 layer farms and one broiler farm near Ulaanbaatar city and isolated 25 Campylobacter jejuni and 6 Campylobacter coli isolates. All isolates were resistant to tetracycline, and 3 C. coli isolates were resistant to erythromycin. The C. coli isolates possessed either the erm(B) gene or nucleotide substitution at nt 2,075 of 23S rDNA, both of which are known to be associated with erythromycin resistance. Sixteen of the 31 C. jejuni/C. coli isolates (51.6%) were resistant to nalidixic acid and fluoroquinolones. All the fluoroquinolone-resistant isolates possessed amino acid substitution from threonine to isoleucine at codon 86 (nucleotide substitution: ACA to ATA). Multilocus sequence typing and phylogenetic analyses showed a variation in C. jejuni/C. coli in chickens in Mongolia. In addition, some of the C. jejuni isolates seemed to be phylogenetically close to isolates in Asian and Oceanian countries. This is the first report on the characterization of antimicrobial resistance of Campylobacter spp. in farm animals in Mongolia and is valuable for implementation of measures for a prudent use of antimicrobials in farm animals.
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Antiinfecciosos , Infecciones por Campylobacter , Campylobacter coli , Campylobacter jejuni , Campylobacter , Animales , Antibacterianos/farmacología , Antiinfecciosos/farmacología , Campylobacter/genética , Infecciones por Campylobacter/epidemiología , Infecciones por Campylobacter/veterinaria , Campylobacter coli/genética , Pollos , Farmacorresistencia Bacteriana , Pruebas de Sensibilidad Microbiana/veterinaria , Mongolia/epidemiología , FilogeniaRESUMEN
DNA polymerase zeta (Polζ) is a heterotetramer composed of the catalytic subunit Rev3l, Rev7 and two subunits of Polδ (PolD2/Pol31 and PolD3/Pol32), and this polymerase exerts translesion DNA synthesis (TLS) in yeast. Because Rev3l knockout results in embryonic lethality in mice, the functions of Polζ need further investigation in vivo. Then, we noted the two facts that substitution of leucine 979 of yeast Rev3l with methionine reduces Polζ replication fidelity and that reporter gene transgenic rodents are able to provide the detailed mutation status. Here, we established gpt delta mouse knocked in the constructed gene encoding methionine instead of leucine at residue 2610 of Rev3l (Rev3l L2610M gpt delta mice), to clarify the role of Polζ in TLS of chemical-induced bulky DNA adducts in vivo. Eight-week-old gpt delta mice and Rev3l L2610M gpt delta mice were treated with benzo[a]pyrene (BaP) at 0, 40, 80, or 160 mg/kg via single intraperitoneal injection. At necropsy 31 days after treatment, lungs were collected for reporter gene mutation assays. Although the gpt mutant frequency was significantly increased by BaP in both mouse genotypes, it was three times higher in Rev3l L2610M gpt delta than gpt delta mice after treatment with 160 mg/kg BaP. The frequencies of G:C base substitutions and characteristic complex mutations were significantly increased in Rev3l L2610M gpt delta mice compared with gpt delta mice. The BaP dose-response relationship suggested that Polζ plays a central role in TLS when protective mechanisms against BaP mutagenesis, such as error-free TLS, are saturated. Overall, Polζ may incorporate incorrect nucleotides at the sites opposite to BaP-modified guanines and extend short DNA sequences from the resultant terminal mismatches only when DNA is heavily damaged.
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Benzo(a)pireno/toxicidad , Daño del ADN/efectos de los fármacos , Reparación del ADN/efectos de los fármacos , Replicación del ADN/efectos de los fármacos , ADN/metabolismo , Mutagénesis , Alanina Transaminasa/genética , Animales , Dominio Catalítico , Aductos de ADN/metabolismo , ADN Polimerasa Dirigida por ADN/fisiología , Femenino , Pulmón/fisiología , Masculino , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Ratones NoqueadosRESUMEN
Although gpt delta rats, as reporter gene-transgenic rats, were originally developed for in vivo mutation assays, they have also been used to evaluate chemical carcinogenesis and comprehensive toxicity. Therefore, it is necessary to accumulate background data on carcinogenicity and general toxicity in gpt delta rats. Here, we investigated the background data of 110-week-old male and female F344 gpt delta rats and wild-type rats. There was no effect of reporter gene transfection on animal survival rates and body weights during the experiment. The relative weight of male gpt delta rat adrenals was significantly higher than that of wild-type rats, possibly due to the higher incidence of pheochromocytoma. There were no intergenotype differences in the incidence of nonneoplastic lesions in both sexes, including chronic progressive nephropathy and focus of cellular alteration in the liver, which had a higher incidence in both genotypes. Additionally, the significantly higher incidence of adrenal pheochromocytoma in male gpt delta rats than that in wild-type rats was likely incidental because of the lack of differences in the incidences of preneoplastic (male and female) and neoplastic (female) adrenal lesions in both genotypes. Other neoplastic lesions in both sexes showed no intergenotype differences in incidence rates, although large granular lymphocytic leukemia in the spleen and Leydig cell tumors in the testes of males showed higher incidence rates. Overall, there were no effects of reporter gene transfection on the spectrum of spontaneous lesions in F344 gpt delta rats, thus supporting their applicability in evaluating chemical toxicity and carcinogenicity.
RESUMEN
Itai-itai (Japanese, "It hurts! It hurts!") disease (IID), a form of osteomalacia, can be induced in ovariectomized rats by long-term administration of cadmium (Cd). This IID rat model shows severe anemia, severe nephropathy, and osteomalacia accompanied by iron (Fe) deposition at the mineralization front. We characterized the pathogenesis of Cd-induced bone lesions by investigating the relationship between Fe deposition and osteoid tissue formation in ovariectomized rats. The rats were injected with CdCl2 (0.5 mg/kg) for 70 weeks, with or without co-injection of erythropoietin (EPO) for varying lengths of time to elucidate whether EPO prevents and/or cures anemia, and, with the restoration from anemia, lessens the osteoid tissue formation. Necropsies were performed at 25, 50, or 70 weeks. Fe deposition at the mineralization front of bone was found at 50 weeks and increased thereafter. Animals injected with EPO showed decreased Fe deposition, although there was no relation between EPO administration and osteoid formation in the femur. Because the increase in bone lesion severity was independent of the amount of Fe deposition, we suggest that Fe deposition is not involved in the etiology of Cd-induced femoral bone lesions.
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Cadmio/toxicidad , Hierro/metabolismo , Osteomalacia/inducido químicamente , Animales , Cadmio/administración & dosificación , Femenino , Fémur/efectos de los fármacos , Fémur/patología , Osteomalacia/patología , Ovariectomía/efectos adversos , Ratas , Ratas Sprague-DawleyRESUMEN
Glutathione S-transferase placental form-positive (GST-P+) foci are markers of preneoplastic lesions in rat hepatocarcinogenesis. Our previous studies using reporter gene transgenic rats showed that furan, a hepatocarcinogen in rodents, rapidly induces the formation of GST-P+ foci after short exposure without reporter gene mutation. We hypothesized that GST-P+ foci induced by furan may have biological characteristics different from those induced by diethylnitrosamine (DEN), a genotoxic hepatocarcinogen. Accordingly, we compared the cell kinetics of GST-P+ foci after cessation of DEN treatment and performed comprehensive gene expression in DEN- or furan-induced GST-P+ foci. The number and area of DEN-induced GST-P+ foci were increased after cessation of treatment, whereas furan decreased these parameters. Size distribution analysis showed that large furan-induced GST-P+ foci disappeared after cessation of treatment. Hierarchical cluster analysis showed that all samples from GST-P+ foci induced by furan were separated from those induced by DEN. SOX9 expression was upregulated in furan-induced GST-P+ foci and was detected by immunohistochemistry in large furan-induced GST-P+ foci. Our results indicated that large furan-induced GST-P+ foci were quite different from DEN-induced GST-P+ foci at the molecular and cellular levels. And one of the properties of disappearing large GST-P+ foci were characterized by inclusion of hepatocytes expressing SOX9.
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Neoplasias Hepáticas Experimentales , Lesiones Precancerosas , Animales , Dietilnitrosamina , Femenino , Furanos/toxicidad , Expresión Génica , Glutatión Transferasa/genética , Glutatión Transferasa/metabolismo , Cinética , Hígado/metabolismo , Placenta/metabolismo , Embarazo , RatasRESUMEN
Canine parvovirus type 2 (CPV-2) causes a highly contagious and fatal disease, developing into acute hemorrhagic enteritis and myocarditis, in dogs. CPV-2 has evolved, generating antigenic variants CPV-2a/2b/2c that are globally distributed. However, investigating molecular characterization of CPV-2 among dog populations in Mongolia has been limited. Herein, 42 stool samples were collected from dogs with clinical signs of infection, and conventional PCR assays were employed to detect CPV-2 in 23. Our results indicated that during 2016-2018, the new CPV-2a and 2c subtypes were detected in 34.7% of the samples, and the new CPV-2b subtype was detected in 30.4% of samples. VP2 protein sequence analysis and next-generation sequencing of the complete viral genome confirmed these antigenic types. However, sequence analysis indicated new and unreported mutations, Pro580Thr, and Tyr584His in the CPV-2c subtype. From a PCR-positive sample, CPV-2c was successfully isolated, and we performed an immunofluorescence assay for antigen detection. Additionally, we performed genetic characterization and phylogenetic analysis to investigate genetic diversity among isolates from the region, resulting in high CPV-2 genetic diversity in the Mongolian dog population. Striking similarities were also observed between sequences of the strains isolated from Mongolia and China over a similar time span.
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Enfermedades de los Perros/virología , Infecciones por Parvoviridae/veterinaria , Parvovirus Canino/genética , Secuencia de Aminoácidos , Animales , Enfermedades de los Perros/epidemiología , Perros , Mongolia/epidemiología , Infecciones por Parvoviridae/virología , Filogenia , Vigilancia de la Población , Proteínas ViralesRESUMEN
Cell proliferation plays a key role in fixing mutations induced by DNA damage. We clarified whether this phenomenon occurred after combined treatment with chemicals in food. The effects of antibiotic flumequine (FL), a residue of veterinary medicinal products in foodstuffs, on mutagenicity in the liver were examined in mice treated with estragole (ES), a natural food flavouring compound. Gpt delta mice were orally administered 10 or 100â¯mg/kg/day ES and simultaneously fed a diet containing 0.4% FL for 4 weeks. Proliferating cell nuclear antigen-positive cells and cell cycle-related genes were additively increased in the livers of combined treatment groups as compared with high-dose ES or FL groups. Mutant frequencies (MFs) in gpt after cotreatment with low-dose ES and FL were significantly increased, although treatment with ES alone increased MFs only in the high-dose group. Sult1a1 mRNA levels were unchanged after FL treatment. Liquid chromatography with tandem-mass spectrometry analysis showed that FL did not affect the amount of ES-specific DNA adducts in the livers, indicating that FL treatment did not influence metabolic pathways of ES. Thus, enhancement of the mutagenic potential of a chemical by chemical-induced cell proliferation may occur as a result of the combined effects of chemicals in food.
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ADN/química , ADN/efectos de los fármacos , Alimentos , Hígado/efectos de los fármacos , Pruebas de Mutagenicidad , Mutación , Animales , Peso Corporal , Proliferación Celular/efectos de los fármacos , Citocinas/genética , Citocinas/metabolismo , Daño del ADN , Mediadores de Inflamación/metabolismo , Hígado/patología , Ratones , Tamaño de los ÓrganosRESUMEN
Sulfotransferase 1A (SULT1A) expression is lower in the liver of humans than that of rodents. Therefore, species differences should be taken into consideration when assessing the risk of rodent hepatocarcinogens metabolically activated by SULT1A in humans. Although some renal carcinogens require SULT1A-mediated activation, it is unclear how SULT1A activity in the liver affects renal carcinogens. To explore the effects of SULT1A activity in the liver on genotoxicity induced by SULT1A-activated renal carcinogens, B6C3F1 mice or gpt delta mice of the same strain background were given lucidin-3-O-primeveroside (LuP), a hepatic and renal carcinogen of rodents, for 4 or 13 weeks, respectively, and pentachlorophenol (PCP) as a liver-specific SULT inhibitor, was given from 1 week before LuP treatment to the end of the experiment. A 4 week exposure of LuP induced lucidin-specific DNA adduct formation. The suppression of Sult1a expression was observed only in the liver but not in the kidneys of PCP-treated mice, but co-administration of PCP suppressed LuP-induced DNA adduct formation in both organs. Thirteen-week exposure of LuP increased mutation frequencies and cotreatment with PCP suppressed these increases in both organs. Given that intact levels of SULT activity in the liver were much higher than in the kidneys of rodents, SULT1A may predominantly activate LuP in the liver, consequently leading to genotoxicity not only in the liver but also in the kidney. Thus, species differences should be considered in human risk assessment of renal carcinogens activated by SULT1A as in the case of the corresponding liver carcinogens.
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Antraquinonas/toxicidad , Disacáridos/toxicidad , Colorantes de Alimentos/toxicidad , Riñón/efectos de los fármacos , Hígado/enzimología , Sulfotransferasas/antagonistas & inhibidores , Animales , Inhibidores Enzimáticos/farmacología , Riñón/patología , Masculino , Ratones , Ratones Endogámicos , Pentaclorofenol/farmacología , Sulfotransferasas/genéticaRESUMEN
Long-term exposure to piperonyl butoxide (PBO) induces multiple nodular masses along with hepatocellular tumors in the liver of mice. The histopathological features of the nodules led to our diagnosis of nodular regenerative hepatocellular hyperplasia (NRH). However, because of the lack of data on the biological characteristics of NRH, whether this lesion is truly nonneoplastic remains unknown. In this study, the molecular characteristics of NRH were compared with those of hepatocellular adenoma (HCA) by global gene expression analysis. Six-week-old male ICR mice were fed a diet containing 6,000 ppm PBO for 43 weeks to induce NRH and HCA development. Complementary DNA microarray analysis was performed using messenger RNA extracted from NRH and HCA frozen sections collected by laser microdissection. Hierarchical cluster analysis showed that all NRH samples clustered together but were separate from the HCA cluster. Pathway analysis revealed activation of the cell cycle and Delta-Notch signaling in both lesions, but the latter was more upregulated in HCA. Downregulation of cytochrome p450 enzymes was observed in NRH, but not in HCA. These results imply that NRH differs from HCA in terms of not only morphological but also molecular characteristics.
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Adenoma de Células Hepáticas/genética , Proliferación Celular/efectos de los fármacos , Neoplasias Hepáticas Experimentales/genética , Hígado/patología , Butóxido de Piperonilo/toxicidad , Transcriptoma/efectos de los fármacos , Adenoma de Células Hepáticas/inducido químicamente , Adenoma de Células Hepáticas/patología , Animales , Diagnóstico Diferencial , Hiperplasia , Hígado/efectos de los fármacos , Neoplasias Hepáticas Experimentales/inducido químicamente , Neoplasias Hepáticas Experimentales/patología , Masculino , Ratones Endogámicos ICR , Análisis de Secuencia por Matrices de Oligonucleótidos , Patología MolecularRESUMEN
Despite its antimicrobial activity, nitrofurantoin (NFT) is a renal carcinogen in rats. Oxidative stress induced by reduction of the nitro group of NFT may contribute to its genotoxicity. This is supported by our recent results indicating that the structure of the nitrofuran plays a key role in NFT-induced genotoxicity, and oxidative DNA damage is involved in renal carcinogenesis. Nuclear factor erythroid 2-related factor 2 (NRF2) regulates cellular responses to oxidative stress. To clarify the role of oxidative stress in the chemical structure-related genotoxic mechanism of NFT, we performed reporter gene mutation assays for NFT and 5-nitro-2-furaldehyde (NFA) using Nrf2-proficient and Nrf2-deficient gpt delta mice. NFT administration for 13 weeks resulted in a significant increase in 8-hydroxydeoxyguanosine (8-OHdG; a marker of oxidative stress) and gpt mutant frequency only in the kidneys of Nrf2-/- mice. The mutation spectrum, characterized by increased substitutions at guanine bases, suggested that oxidative stress is involved in NFT-induced genotoxicity. However, NFA did not increase the mutation frequency in the kidneys, despite the increased 8-OHdG in NFA-treated Nrf2-/- mice. Thus, it is unlikely that oxidative stress is involved in the genotoxic mechanism of NFA. These results imply that nitro reduction plays a key role in the genotoxicity of NFT, but the lack of a role of oxidative stress in the genotoxicity of NFA indicates a potential role of side chain interactions in oxidative stress caused by nitro reduction. These findings provide a basis for the development of safe nitrofurans.
RESUMEN
Oxidative stress is well known as a key factor of chemical carcinogenesis. However, the actual role of oxidative stress in carcinogenesis, such as oxidative stress-related in vivo mutagenicity, remains unclear. It has been reported that 8-hydroxydeoxyguanosine (8-OHdG), an oxidized DNA lesion, might contribute to chemical carcinogenesis. Potassium bromate (KBrO3) and nitrofurantoin (NFT) are known as renal carcinogens in rats. Our previous studies showed an increase in mutant frequencies accompanied by an increased level of 8-OHdG in the kidneys of rodents following KBrO3 or NFT exposure. Furthermore, KBrO3 and NFT induced different types of gene mutations. Thus, in the present study, we performed reporter gene mutation assays and 8-OHdG measurements following KBrO3 or NFT exposure using Nrf2-proficient and Nrf2-deficient mice to clarify the relationship between KBrO3- or NFT-induced oxidative stress and subsequent genotoxicity. Administration of 1,500 ppm of KBrO3 in drinking water resulted in an increase in deletion mutations accompanied by an increase in 8-OHdG level, and administration of 2,500 ppm of NFT in diet induced an increase in guanine base substitution mutations without elevation of the 8-OHdG level in Nrf2-deficient mice. These results demonstrated that the formation of 8-OHdG, which resulted from the oxidizing potential of KBrO3, was directly involved in the increase in deletion mutations, although factors related to oxidative stress other than 8-OHdG might be crucial for NFT-induced guanine base substitution mutations. The present study provides new insight into oxidative stress-related in vivo mutagenicity.
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Osmotic nephrosis, a disease caused by intravenous infusion of various fluids such as hypertonic sucrose and isotonic polysaccharide-based plasma volume expanders, exhibits specific histopathological features, including vacuolated and swollen proximal tubules, ie, "clear tubules". Pre-existing kidney injury exacerbates this condition, resulting in major clinical problems. However, the underlying mechanisms are unclear. Animal models often yield results that are directly translatable to humans. Therefore, in this study, we performed detailed histopathological analyses of the formation of clear tubules in rats treated with gentamicin or ischemia/reperfusion (IR) operation followed by dextran administration. The results showed that clear tubules may originate from regenerative tubules. Additionally, we classified regenerative tubules into 3 categories based on their development, with a particular focus on the middle and late stages. Comprehensive microarray and real-time polymerase chain reaction analyses of mRNA extracted from regenerative tubules at each stage using laser microdissection revealed that regenerative tubules in the middle stage showed an imbalance between dextran absorption and metabolism, resulting in accumulation of dextran, particularly in the cytoplasm of the tubules. Overall, our findings demonstrated that clear tubules originated from regenerated tubules and that tubules at the middle stage became clear tubules because of an imbalance during their development. This could explain why osmotic nephrosis is exacerbated in the presence of kidney lesions.
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Lesión Renal Aguda/patología , Túbulos Renales Proximales/patología , Nefrosis/patología , Daño por Reperfusión/patología , Lesión Renal Aguda/complicaciones , Lesión Renal Aguda/metabolismo , Animales , Dextranos/metabolismo , Dextranos/toxicidad , Modelos Animales de Enfermedad , Progresión de la Enfermedad , Femenino , Gentamicinas/metabolismo , Gentamicinas/toxicidad , Túbulos Renales Proximales/metabolismo , Masculino , Nefrosis/etiología , Nefrosis/metabolismo , Ratas Endogámicas F344 , Ratas Sprague-Dawley , Daño por Reperfusión/complicaciones , Daño por Reperfusión/metabolismoRESUMEN
Lead (Pb) and cadmium (Cd) are toxic metals that exist ubiquitously in the environment. Children in polluted areas are particularly vulnerable to metal exposure, where clinical signs and symptoms could be nonspecific. Absorbed metals are excreted primarily in urine and reflect exposure from all sources. We analyzed Pb and Cd concentrations in blood, feces and urine of children from polluted townships near a lead-zinc mine in Kabwe, Zambia, to determine concurrent childhood exposure to the metals. Moreover, the study determined the Pb and Cd relationships among urine, feces and blood as well as accessed the potential of urine and fecal analysis for biomonitoring of Pb and Cd exposure in children. Fecal Pb (up to 2252â¯mg/kg, dry weight) and urine Pb (up to 2914⯵g/L) were extremely high. Concentrations of Cd in blood (Cd-B) of up to 7.7⯵g/L, fecal (up to 4.49â¯mg/kg, dry weight) and urine (up to 18.1⯵g/L) samples were elevated. metal levels were higher in younger children (0-3 years old) than older children (4-7). Positive correlations were recorded for Pb and Cd among blood, urine and fecal samples whereas negative correlations were recorded with age. These findings indicate children are exposed to both metals at their current home environment. Moreover, urine and feces could be useful for biomonitoring of metals due to their strong relationships with blood levels. There is need to conduct a clinical evaluation of the affected children to fully appreciate the health impact of these metal exposure.
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Cadmio/análisis , Monitoreo del Ambiente/métodos , Contaminantes Ambientales/análisis , Heces/química , Intoxicación por Metales Pesados/orina , Plomo/análisis , Urinálisis/métodos , Peso Corporal , Niño , Preescolar , Femenino , Humanos , Lactante , Masculino , ZambiaRESUMEN
The genetic characterization and actual prevalence of EIAV in Mongolian horse in the disease endemic region is currently unknown. Here, 11 of 776 horse serum samples from four Mongolian provinces tested positive on agar gel immunodiffusion test. Genomic DNA extracted from all seropositive samples was subjected to nested PCR assay. Among these, three samples tested positive with nested PCR assay and were identified by sequencing analysis based on long termination repeat and tat gene of the virus. Two of the three sequences were identical, with 94.0% identity with the third. These two independent Mongolian EIAV sequences were retained functional motifs, with no dramatic changes but some variability in the U5 region; they were clustered with genotypes from European countries but not with those from China, U.S.A., or Japan.
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Anemia Infecciosa Equina/virología , Virus de la Anemia Infecciosa Equina/genética , Virus de la Anemia Infecciosa Equina/aislamiento & purificación , Animales , ADN Viral , Anemia Infecciosa Equina/epidemiología , Variación Genética , Genoma Viral , Caballos , Mongolia/epidemiología , Reacción en Cadena de la Polimerasa/veterinaria , Análisis de Secuencia de ADN , Estudios SeroepidemiológicosRESUMEN
Protein phosphatase 2A (PP2A) is a serine-threonine phosphatase that regulates cell signaling pathways. Its inactivation is correlated with tumor malignancy, possibly due to the effects on cell differentiation and malignant cell transformation. Therefore, it has been noted that PP2A could be a promising target for cancer therapy. In our previous study of the hepatocarcinogen estragole (ES), cell proliferation may be required to convert ES-specific DNA adducts to mutations. To explore the trigger for cell proliferation, gpt delta rats were administered ES by gavage at doses of 3, 30 and 300mg/kg/day for 4weeks. ES-induced cell proliferation and gene mutations were observed at only the high dose whereas ES-specific DNA adducts were detected in a dose-dependent manner. Western blot analyses revealed activation of the Akt and ERK pathways without activation of upstream regulators, such as c-Raf, PKC and, PI3K. Phosphorylation of the PP2A C subunit at Tyr307 was found along with phosphorylation of Src. The overall data might imply that PP2A inactivation is responsible for cell cycle progression through activation of the Akt and ERK pathways at high doses of ES. Based on γ-H2AX immunohistochemistry and Western blot analysis for Rad51 protein, the resultant mutation spectra showed large deletion mutations that might result from double strand breaks of DNA. Thus, it is likely that inactivation of PP2A resulted in acceleration and exacerbation of gene mutations. We conclude that PP2A might contribute to an early stage of chemical carcinogenesis, suggesting that PP2A could be a molecular target of primary cancer prevention.