Effects of Fluoride on Submandibular Glands of Mice: Changes in Oxidative Biochemistry, Proteomic Profile, and Genotoxicity

Although fluoride (F) is well-known to prevent dental caries, changes in cell processes in different tissues have been associated with its excessive exposure. Thus, this study aimed to evaluate the effects of F exposure on biochemical, proteomic, and genotoxic parameters of submandibular glands. Twenty one old rats (n = 30) were allocated into three groups: 60 days administration of drinking water containing 10 mgF/L, 50 mgF/L, or only deionized water (control). The submandibular glands were collected for oxidative biochemistry, protein expression profile, and genotoxic potential analyses. The results showed that both F concentrations increased the levels of thiobarbituric acid–reactive substances (TBARS) and reduced glutathione (GSH) and changed the proteomic profile, mainly regarding the cytoskeleton and cellular activity. Only the exposure to 50 mgF/L induced significant changes in DNA integrity. These findings reinforce the importance of continuous monitoring of F concentration in drinking water and the need for strategies to minimize F intake from other sources to obtain maximum preventive/therapeutic effects and avoid potential adverse effects

Effects of long-term fluoride exposure are associated with oxidative biochemistry impairment and global proteomic modulation, but not genotoxicity, in parotid glands of mice

Fluoride has become widely used in dentistry because of its effectiveness in caries control. However, evidence indicates that excessive intake interferes with the metabolic processes of different tissues. Thus, this study aimed to investigate the effects of long-term exposure to F on the parotid salivary gland of mice, from the analysis of oxidative, proteomic and genotoxic parameters. The animals received deionized water containing 0, 10 or 50 mg/L of F, as sodium fluoride, for 60 days. After, parotid glands were collected for analysis of oxidative biochemistry, global proteomic profile, genotoxicity assessment and histopathological analyses. The results revealed that exposure to fluoride interfered in the biochemical homeostasis of the parotid gland, with increased levels of thiobarbituric acid reactive species and reduced glutathione in the exposed groups; as well as promoted alteration of the glandular proteomic profile in these groups, especially in structural proteins and proteins related to oxidative stress. However, genotoxic assessment demonstrated that exposure to fluoride did not interfere with DNA integrity in these concentrations and durations of exposure. Also, it was not observed histopathological alterations in parotid gland

Análises cromossômicas no aconselhamento genético de pacientes com alterações no desenvolvimento e congênitas em Belém, estado do Pará, Brasil: um estudo retrospectivo de 17 anos

Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Laboratório de Cultura de Tecidos e Citogenética. Ananindeua, PA, Brasil.Universidade Federal do Pará. Pró-Reitoria de Pesquisa e Pós-Graduação. Programa de Bolsas de Iniciação Científica. Belém, PA, Brasil.Universidade Federal do Pará. Instituto de Ciências Biológicas. Laboratório de Biologia Molecular Francisco Mauro Salzano. Belém, PA, Brasil.Universidade Federal do Pará. Instituto de Ciências Biológicas. Laboratório de Citogenética Humana e Clínica. Belém, PA, Brasil.Universidade Federal do Pará. Instituto de Ciências Biológicas. Laboratório de Citogenética Humana e Clínica. Belém, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Laboratório de Cultura de Tecidos e Citogenética. Ananindeua, PA, Brasil / Universidade Federal do Pará. Instituto de Ciências Exatas e Naturais. Faculdade de Ciências Naturais. Belém, PA, Brasil.Universidade Federal do Pará. Instituto de Ciências Biológicas. Laboratório de Citogenética Humana e Clínica. Belém, PA, Brasil.OBJECTIVE: To present a retrospective study covering 17 years of referral of patients to a public clinical cytogenetic service in Belém, Pará State, located in the Brazilian Amazon. MATERIALS AND METHODS: This study was based on a retrospective survey conducted from 1997 to 2014, considering registered chromosome G-banding results and relating them to information collected during patient evaluation. RESULTS: From a total of 1,580 patients, 730 (46.2%) had chromosomal abnormalities, of which 637 (87.3%) showed numerical alterations. Abnormalities involving autosomal chromosomes were more frequent, 524/730 (71.8%), while alterations in sex chromosomes comprised 28.2% (206). Down's syndrome was the most frequent, 424 (58.1%) of cases, followed by 175 (24.0%) cases of Turner's syndrome, and 25 (3.4%) of Klinefelter's syndrome. Patients with sex chromosome abnormalities were referred at a more advanced age when compared with those having autosomal chromosome abnormalities, with peaks around 11–15 years old (30.1% of cases) and 0–6 months old (40.5%), respectively. CONCLUSION: These findings are very similar to other studies and draw attention to public measures to improve both the quality with regard to diagnosis and the subsequent care of the patient

Copy number alterations in papillary thyroid carcinomas: does loss of SESN2 have a role in age-related different prognoses?

This study was partially supported by the Ministry of Health through the Evandro Chagas Institute official budget and by CNPq (process 307382/2019-2)Federal University of Pará. Institute of Biological Sciences. Neuroscience and Cell Biology Pos-Graduation Program. Belém, PA, Brazil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Ananindeua, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Laboratório de Cultura de Tecidos e Citogenética. Ananindeua, PA, Brasil.Federal University of Pará. Institute of Biological Sciences. Laboratory of Human Cytogenetics. Belém, PA, Brazil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Laboratório de Cultura de Tecidos e Citogenética. Ananindeua, PA, Brasil.Federal University of Ceará. Drug Research and Development Center. Pharmacogenetics Laboratory. Fortaleza, CE, Brazil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Laboratório de Cultura de Tecidos e Citogenética. Ananindeua, PA, Brasil / Federal University of Pará. Institute of Exact and Natural Sciences. Belém, PA, Brazil.Background/Aim: Thyroid cancer is the only tumor in which age is an important prognostic factor. In papillary thyroid carcinomas (PTC), 45 years of age seems to be a key point that divides adult patients into two groups, with different clinical features. The aim of the study was to perform a microarray-based analysis in two groups of patients (<45 and ≥45 years old), in order to verify the occurrence of specific copy number alterations (CNAs) that could be associated to different patient behaviors associated with age. Patients and Methods: In order to search and compare genomic alterations that may be related to age, we evaluated the occurrence of CNAs in the genome of 24 PTC samples, divided in two groups (<45 and ≥45 years old). Results: We identified only one region showing a statistically significant difference between the groups (p=0.00357): a deletion of approximately 537 kps in 1p35.3., which was more frequent in patients aged 45 years or older. This is the region where, among others, the gene SESN2 is located, which is activated under oxidative stress and plays an antioxidant role, in addition to protecting the genetic material from damage generated by reactive oxygen species (ROS). Conclusion: This is the first time that a CNA involving the deletion of the SESN2 gene is associated with papillary thyroid carcinomas, particularly in patients aged 45 years and older, indicating that this deletion would lead to a more malignant and prominent tumoral behavior associated to a worst prognosis

Fluoride exposure during pregnancy and lactation triggers oxidative stress and molecular changes in hippocampus of offspring rats

This work was supported by Pro-Reitoria ´ de Pesquisa da UFPA (PROPESP, UFPA, Brazil), Brazilian National Council for Scientific and Technological Development (CNPq, Grant n. 435093/2018-5) and Coordenaçao ˜ de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)- Finance Code 001.Federal University of Pará. Institute of Biological Sciences. Laboratory of Functional and Structural Biology. Belém, PA, Brazil.Federal University of Pará. Institute of Biological Sciences. Laboratory of Functional and Structural Biology. Belém, PA, Brazil.Federal University of Pará. Institute of Biological Sciences. Laboratory of Functional and Structural Biology. Belém, PA, Brazil.Federal University of Pará. Institute of Biological Sciences. Laboratory of Functional and Structural Biology. Belém, PA, Brazil.University of São Paulo. Bauru Dental School. Department of Biological Sciences. Bauru, SP, Brazil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Laboratório de Cultura Celular e Citogenética. Ananindeua, PA, Brasil.University of São Paulo. Bauru Dental School. Department of Biological Sciences. Bauru, SP, Brazil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Laboratório de Cultura Celular e Citogenética. Ananindeua, PA, Brasil.Federal University of Pará. Institute of Biological Sciences. Laboratory of Molecular Pharmacology. Belém, PA, Brazil.Federal University of Pará. Institute of Biological Sciences. Laboratory of Functional and Structural Biology. Belém, PA, Brazil.Long-term exposure to high concentrations of fluoride (F) can damage mineralized and soft tissues such as bones, liver, kidney, intestine, and nervous system of adult rats. The high permeability of the blood–brain barrier and placenta to F during pregnancy and lactation may be critical to neurological development. Therefore, this study aimed to investigate the effects of F exposure during pregnancy and lactation on molecular processes and oxidative biochemistry of offspring rats’ hippocampus. Pregnant Wistar rats were randomly assigned into 3 groups in accordance with the drinking water received: G1 – deionized water (control); G2 – 10 mg/L of F and G3 – 50 mg/L of F. The exposure to fluoridated water began on the first day of pregnancy and lasted until the 21st day of breastfeeding (when the offspring rats were weaned). Blood plasma samples of the offspring rats were collected to determine F levels. Hippocampi samples were collected for oxidative biochemistry analyses through antioxidant capacity against peroxyl (ACAP), lipid peroxidation (LPO), and nitrite (NO2 − ) levels. Also, brainderived neurotrophic factor (BDNF) gene expression (RT-qPCR) and proteomic profile analyses were performed. The results showed that exposure to both F concentrations during pregnancy and lactation increased the F bioavailability, triggered redox imbalance featured by a decrease of ACAP, increase of LPO and NO2 − levels, BDNF overexpression and changes in the hippocampus proteome. These findings raise novel questions regarding potential repercussions on the hippocampus structure and functioning in the different cognitive domains

DNA damage and proteomic profile changes in rat salivary glands after chronic exposure to inorganic mercury

This study was financed in part by the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior–Brasil (CAPES)–Financing Code 001.Federal University of Pará. Institute of Biological Sciences. Laboratory of Functional and Structural Biology. Belém, PA, Brazil.Federal University of Pará. Institute of Biological Sciences. Laboratory of Functional and Structural Biology. Belém, PA, Brazil.Federal University of Pará. Institute of Biological Sciences. Laboratory of Functional and Structural Biology. Belém, PA, Brazil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Ananindeua, PA, BrasilMinistério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Laboratório de Cultura Celular e Citogenética. Ananindeua, PA, BrasilUniversity of São Paulo. Bauru School of Dentistry. Department of Biological Sciences. Bauru, SP, BrazilUniversity of São Paulo. Bauru School of Dentistry. Department of Biological Sciences. Bauru, SP, BrazilMinistério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Laboratório de Cultura Celular e Citogenética. Ananindeua, PA, BrasilFederal University of Pará. Institute of Biological Sciences. Laboratory of Molecular Pharmacology. Belém, PA, BrazilFederal University of Pará. Institute of Biological Sciences. Laboratory of Functional and Structural Biology. Belém, PA, BrazilMercury (Hg) is a toxic metal that became a public health problem due to environmental contamination caused by anthropogenic activity. In this sense, oral homeostasis can undergo changes due to the toxic effects of metal on the salivary glands. Therefore, our objective was to investigate the proteomic and genotoxic changes in salivary glands after exposure to inorganic mercury (IHg). Forty Wistar rats that were divided into a control group, which received distilled water, and an exposed group, which received 0.375 mg/kg of mercury chloride for 45 days via orogastric gavage. After that, the animals were euthanized, and the parotid and submandibular glands were collected for analysis of the genotoxic effects, using the comet assay and proteome global profile assessment. The results showed that IHg promoted damage to cellular DNA associated with proteomic changes that showed events such as oxidative stress, mitochondrial dysfunction, changes in the cytoskeleton, and apoptosis. Therefore, these findings show a profile of molecular changes due to the interactions of IHg with several proteins and mechanisms inherent to the cell, which consequently may result in dysfunction of the salivary glands and impaired homeostasis of the oral cavity

Chronic methylmercury exposure causes spinal cord impairment: proteomic modulation and oxidative stress

Conselho Nacional de Desenvolvimento Científico e Tecnológico – Brasil (CNPq) and Coordenação de Aperfeiçoamento de Pessoal de Nível Superior Brasil (CAPES) - Finance Code 001; Universidade Federal do Pará (PROPESP, UFPA, Brazil)Federal University of Pará. Institute of Biological Sciences. Laboratory of Functional and Structural Biology. Belém, PA, Brazil.Federal University of Pará. Institute of Biological Sciences. Laboratory of Functional and Structural Biology. Belém, PA, Brazil.Federal University of Pará. Institute of Biological Sciences. Laboratory of Functional and Structural Biology. Belém, PA, Brazil.Federal University of Pará. Institute of Biological Sciences. Laboratory of Functional and Structural Biology. Belém, PA, Brazil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Ananindeua, PA, Brasil.Federal University of Pará. Institute of Biological Sciences. Laboratory of Functional and Structural Biology. Belém, PA, Brazil.University of São Paulo. Department of Biological Sciences. Bauru Dental School. Bauru, SP, Brazil.University of São Paulo. Department of Biological Sciences. Bauru Dental School. Bauru, SP, Brazil.Federal University of Pará. Laboratory of Molecular Pharmacology. Belém, PA, Brazil.Federal University of Pará. Institute of Biological Sciences. Laboratory of Functional and Structural Biology. Belém, PA, Brazil.Methylmercury (MeHg) is considered by the World Health Organization (WHO) as one of the chemicals of greatest public health concern. Although central nervous system (CNS) is the main target organ, the effects over the spinal cord are not well understood, especially in chronic exposure at similar doses to those faced by humans. This study aimed to investigate possible changes on global proteomic profile and oxidative biochemistry status of rats spinal cord, related to the maintenance and balance of the organism functioning, mimicking a human daily exposure by diet (chronic and with relatively low levels). For this, 28 adults male Wistar rats were divided into two groups: MeHg group, which was intoxicated by intragastric gavage with MeHg at a dose of 0.04 mg/kg/day for 60 days, and control group, that received only vehicle. After the exposure period, the spinal cords were collected for evaluation of total mercury levels, proteomic profile, with further bioinformatic overrepresentation analysis (ORA), and oxidative biochemistry, by analyzing the antioxidant capacity against peroxyl radicals (ACAP), lipid peroxidation (LPO), nitrite levels, measurement of Trolox Equivalent Antioxidant Capacity (TEAC) and Reduced Glutathione (GSH). The MeHg exposure increased total mercury levels in spinal cord parenchyma, which increased lipid peroxidation and nitrite levels, and reduced antioxidant status. The proteomic analysis showed several proteins related to biological processes, cellular components and molecular functions. Moreover, according to the ORA analysis, the proteins are involved in processes such as mitochondrial activity, stress response, cytoskeleton and apoptosis. Therefore, we concluded that exposure to low doses of MeHg can activate the oxidative stress pathway and thus, modulate the status of regulation of several important proteins

Identifying novel genetic alterations in pediatric acute lymphoblastic leukemia based on copy number analysis

National Counsel of Technological and Scientific Development (CNPq), grant n° 460185/2014–4 and Amazon Foundation for Research Support (FAPESPA), grant n° PPSUS/2013.Federal University of Pará. Oncology Research Center. Belém, PA, Brazil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Ananindeua, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Ananindeua, PA, Brasil.Octávio Lobo Children’s Cancer Hospital. Belém, PA, Brazil.Octávio Lobo Children’s Cancer Hospital. Belém, PA, Brazil.Federal University of Pará. Oncology Research Center. Belém, PA, Brazil.Federal University of Pará. Oncology Research Center. Belém, PA, Brazil.Federal University of Pará. Oncology Research Center. Belém, PA, Brazil.Federal University of Pará. Oncology Research Center. Belém, PA, Brazil.Copy number variations (CNVs) analysis may reveal molecular biomarkers and provide information on the pathogenesis of acute lymphoblastic leukemia (ALL). We investigated the gene copy number in childhood ALL by microarray and select three new recurrent CNVs to evaluate by real-time PCR assay: DMBT1, KIAA0125 and PRDM16 were selected due to high frequency of CNVs in ALL samples and based on their potential biological functions in carcinogenesis described in the literature. DBMT1 deletion was associated with patients with chromosomal translocations and is a potential tumor suppressor; KIAA0125 and PRDM16 may act as an oncogene despite having a paradoxical behavior in carcinogenesis. This study reinforces that microarrays/aCGH is it is a powerful tool for detection of genomic aberrations, which may be used in the risk stratificatio

Mesenchymal stem cells in dogs with demyelinating leukoencephalitis as an experimental model of multiple sclerosis

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES), Brasil (Finance Code 001).Federal Rural University of Amazonia. Faculty of Veterinary Medicine. Institute of Animal Health and Production. Belém, PA, Brazil.Federal Rural University of Amazonia. Faculty of Veterinary Medicine. Institute of Animal Health and Production. Belém, PA, Brazil.Federal Rural University of Amazonia. Faculty of Veterinary Medicine. Institute of Animal Health and Production. Belém, PA, Brazil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Laboratório de Cultura de Tecidos e Citogenética. Ananindeua, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Laboratório de Cultura de Tecidos e Citogenética. Ananindeua, PA, Brasil.University of São Paulo. Faculty of Veterinary Medicine and Animal Science. Department of Surgery. São Paulo, SP, Brazil.University of São Paulo. Institute of Biomedical Sciences. Department of Microbiology. São Paulo, SP, Brazil.University of São Paulo. Institute of Biomedical Sciences. Department of Microbiology. São Paulo, SP, Brazil.University of São Paulo. Institute of Biomedical Sciences. Department of Microbiology. São Paulo, SP, Brazil.Federal Rural University of Amazonia. Faculty of Veterinary Medicine. Institute of Animal Health and Production. Belém, PA, Brazil.Federal Rural University of Amazonia. Faculty of Veterinary Medicine. Institute of Animal Health and Production. Belém, PA, Brazil.Researchers have used dogs with neurological sequelae caused by distemper as an experimental model for multiple sclerosis, owing to the similarities of the neuropathological changes between distemper virus-induced demyelinating leukoencephalitis and multiple sclerosis in humans. However, little is known about the role of mesenchymal stem cells in treating such clinical conditions. Therefore, we investigated the use of mesenchymal stem cells in four dogs with neurological lesions caused by the distemper virus. During the first year after cellular therapy, the animals did not demonstrate significant changes in their locomotive abilities. However, the intense (Grade V) myoclonus in three animals was reduced to a moderate (Grade IV) level. At one year after the mesenchymal stem cell infusions, three animals regained functional ambulation (Grade I), and all four dogs started to move independently (Grades I and II). In two animals, the myoclonic severity had become mild (Grade III). It was concluded that the use of mesenchymal stem cells could improve the quality of life of dogs with neurological sequelae caused by canine distemper, thus presenting hope for similar positive results in human patients with multiple sclerosis

Piwi like RNA-mediated gene silencing 1 gene as a possible major player in gastric cancer

Fundação Amazônia de Amparo a Estudos e Pesquisa (FAPESPA), No. 174/2014.Universidade Federal do Pará. Núcleo de Pesquisas em Oncologia. Belém, PA, Brazil.Universidade Federal do Pará. Núcleo de Pesquisas em Oncologia. Belém, PA, Brazil.Universidade Federal do Pará. Núcleo de Pesquisas em Oncologia. Belém, PA, Brazil.Universidade Federal do Pará. Núcleo de Pesquisas em Oncologia. Belém, PA, Brazil.Universidade Federal do Pará. Núcleo de Pesquisas em Oncologia. Belém, PA, Brazil.Universidade Federal do Pará. Núcleo de Pesquisas em Oncologia. Belém, PA, Brazil.Universidade Federal do Pará. Núcleo de Pesquisas em Oncologia. Belém, PA, Brazil.Universidade Federal do Pará. Núcleo de Pesquisas em Oncologia. Belém, PA, Brazil.Universidade Federal do Pará. Núcleo de Pesquisas em Oncologia. Belém, PA, Brazil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Laboratório de Cultura de Tecidos e Citogenética. Ananindeua, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Laboratório de Cultura de Tecidos e Citogenética. Ananindeua, PA, Brasil.Universitätsklinikum Jena. Institute of Human Genetics. Jena, Germany.Universitätsklinikum Jena. Institute of Human Genetics. Jena, Germany.Instituto Nacional de Câncer José Alencar Gomes da Silva. Centro de Transplante de Medula Óssea. Laboratório de Célula Tronco. Rio de Janeiro, RJ, Brazil.Instituto Nacional de Câncer José Alencar Gomes da Silva. Centro de Transplante de Medula Óssea. Laboratório de Célula Tronco. Rio de Janeiro, RJ, Brazil.Universidade Federal do Pará. Núcleo de Pesquisas em Oncologia. Belém, PA, Brazil.Universidade Federal do Pará. Núcleo de Pesquisas em Oncologia. Belém, PA, Brazil.AIM: To establish a permanent piwi like RNA-mediated gene silencing 1 (PIWIL1) gene knockout in AGP01 gastric cancer cell line using CRISPR-Cas9 system and analyze phenotypic modifications as well as gene expression alterations. METHODS: CRISPR-Cas9 system used was purchased from Dharmacon GE Life Sciences (Lafayette, CO, United States) and permanent knockout was performed according to manufacturer's recommendations. Wound-healing assay was performed to investigate the effect of PIWIL1 knockout on migration capability of cells and Boyden chamber invasion assay was performed to investigate the effect on invasion capability. For the gene expression analysis, a one-color microarray-based gene expression analysis kit (Agilent Technologies, Santa Clara, CA, United States) was used according to the protocol provided by the manufacturer. RESULTS: PIWIL1 gene knockout caused a significant decrease in AGP01 migration capacity as well as a significant decrease in cell invasiveness. Moreover, functional analysis based on grouping of all differentially expressed mRNAs identified a total of 35 genes (5 up-regulated and 30 down-regulated) encoding proteins involved in cellular invasion and migration. According to current literature, 9 of these 35 genes (DOCK2, ZNF503, PDE4D, ABL1, ABL2, LPAR1, SMAD2, WASF3 and DACH1) are possibly related to the mechanisms used by PIWIL1 to promote carcinogenic effects related to migration and invasion, since their functions are consistent with the changes observed (being up- or down-regulated after knockout). CONCLUSION: Taken together, these data reinforce the idea that PIWIL1 plays a crucial role in the signaling pathway of gastric cancer, regulating several genes involved in migration and invasion processes; therefore, its use as a therapeutic target may generate promising results in the treatment of gastric cancer