title: Evaluation of early changes induced by diuron in the rat urinary bladder using different processing methods for scanning electron microscopy.
abstract: Diuron [3-(3,4-dichlorophenyl)-1,1-dimethylurea] is a substituted urea herbicide carcinogenic to the rat urinary bladder at high dietary levels. The suggested non-genotoxic mode of action (MOA) of diuron encompasses cytotoxicity and necrosis followed by regenerative hyperplasia. Prenecrotic swollen cells as observed under scanning electron microscopy (SEM) have been reported as early morphological alterations, putatively related to diuron cytotoxicity. However, these changes were not observed in a previous SEM study conducted in this laboratory. This study evaluated whether these early alterations are actually due to diuron cytotoxicity or artifacts related to different processing methods used for SEM analysis. Male Wistar rats were fed ad libitum with basal diet, 7.1% sodium saccharin (NaS) or 2.500ppm diuron for seven days or 15 weeks. The urinary bladders were processed for histological and labeling indices examinations and for SEM using two different processing methods. The incidence of simple hyperplasia after 15 weeks of exposure to diuron or to NaS was significantly increased. By SEM, the incidences and severity of lesions were significantly increased in the diuron group independently of exposure time. The different SEM processing methods used allowed for visualization of swollen superficial cells after seven days of diuron exposure. Probably the absence these cells in a previous study was due to the use very few animals. Our results support the hypothesis that the swollen cell is an early key event due to diuron-induced cytotoxicity and is the result of a degenerative process involved in the non-genotoxic carcinogenic mode of action of high doses of diuron.
authors.affiliation: Center for the Evaluation of the Environmental Impact on Human Health (TOXICAM), Department of Pathology, Botucatu Medical School, UNESP - São Paulo State University, Botucatu, São Paulo 18618-970, Brazil. Electronic address: ramarono@gmail.com.
authors.forename: Rafaela Marono
authors.initials: RM
authors.lastname: Fava
authors.affiliation: Center for the Evaluation of the Environmental Impact on Human Health (TOXICAM), Department of Pathology, Botucatu Medical School, UNESP - São Paulo State University, Botucatu, São Paulo 18618-970, Brazil. Electronic address: anaferragut@fmb.unesp.br.
authors.forename: Ana Paula
authors.initials: AP
authors.lastname: Ferragut Cardoso
authors.affiliation: Planitox, Campinas, 13024-230 SP, Brazil. Electronic address: mitscheli@gmail.com.
authors.forename: Mitscheli Sanches
authors.initials: MS
authors.lastname: da Rocha
authors.affiliation: Center for the Evaluation of the Environmental Impact on Human Health (TOXICAM), Department of Pathology, Botucatu Medical School, UNESP - São Paulo State University, Botucatu, São Paulo 18618-970, Brazil. Electronic address: merielen@fmb.unesp.br.
authors.forename: Merielen Garcia
authors.initials: MG
authors.lastname: Nascimento E Pontes
authors.affiliation: Center for the Evaluation of the Environmental Impact on Human Health (TOXICAM), Department of Pathology, Botucatu Medical School, UNESP - São Paulo State University, Botucatu, São Paulo 18618-970, Brazil. Electronic address: decam@fmb.unesp.br.
authors.forename: João Lauro Viana
authors.initials: JLV
authors.lastname: de Camargo
authors.affiliation: Center for the Evaluation of the Environmental Impact on Human Health (TOXICAM), Department of Pathology, Botucatu Medical School, UNESP - São Paulo State University, Botucatu, São Paulo 18618-970, Brazil. Electronic address: mdeolive@fmb.unesp.br.
authors.forename: Maria Luiza Cotrim Sartor
authors.initials: MLCS
authors.lastname: de Oliveira
mesh_terms: D019342:Acetic Acid; D000818:Animals; D049108:Cell Enlargement / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D048429:Cell Size / Q000187:drug effects; D004237:Diuron / Q000633:toxicity*; D005557:Formaldehyde; D005976:Glutaral; D006965:Hyperplasia; D008297:Male; D008855:Microscopy, Electron, Scanning; D010851:Picrates; D017208:Rats, Wistar; D012821:Silanes; D013048:Specimen Handling / Q000379:methods*; D013997:Time Factors; D016707:Tissue Fixation; D001743:Urinary Bladder / Q000187:drug effects* / Q000648:ultrastructure; D019459:Urothelium / Q000187:drug effects* / Q000648:ultrastructure
chemical_list: D010851:Picrates; D012821:Silanes; C520781:hexamethyldisilane; D005557:Formaldehyde; C026239:Bouin's solution; D004237:Diuron; D019342:Acetic Acid; D005976:Glutaral
keywords: Carcinogenesis; Diuron[3-(3,4-dichlorophenyl)-1,1-dimethylurea]; Early change; Rat urinary bladder; Scanning electron microscopy; Swollen cells