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Supplemental material, sj-docx-1-jdr-10.1177_00220345211062475 for Scoping Review of Oral Health-Related Birth Cohort Studies: Toward a Global Consortium by K.G. Peres, G.G. Nascimento, A. Gupta, A. Singh, L. Schertel Cassiano and A.J. Rugg-Gunn in Journal of Dental Research

Jaboticaba Plinia peruviana (Poir.) Govaerts is a Brazilian berry that presents high levels of polyphenols, which may play a key role in preventing cytotoxic and genotoxic effects of harmful agents. Although copper is an essential micronutrient that plays an important role in organisms, high copper concentrations may trigger toxicity to animals and plants. Here, we investigated whether Plinia peruviana hydroalcoholic extract prevents copper-induced cytotoxicity in Allium cepa root cells. Five different anthocyanins and phenolic compounds were identified in Plinia peruviana extract. Importantly, the exposure to 1.53 mg/L copper for 24 h impaired mitotic index, as well as increased mitosis disturbances and triggered DNA damage. Pre-incubation with Plinia peruviana extract (0.25 g/L and 0.75 g/L) for 3 h prevented copper-induced changes in the mitotic index and reduced the number of abnormal cells. In conclusion, we suggest that Plinia peruviana peel extract has protective effects against cellular and genetic disturbances induced by copper.

Several model organisms have been employed to study the impacts of stress on biological systems. Different models of unpredictable chronic stress (UCS) have been established in rodents; however, these protocols are expensive, long-lasting, and require a large physical structure. Our group has recently reported an UCS protocol in zebrafish with several advantages compared to rodent models. We observed that UCS induced behavioral, biochemical, and molecular changes similar to those observed in depressed patients, supporting the translational relevance of the protocol. Considering that a pharmacological assessment is lacking in this zebrafish model, our aim was to evaluate the effects of anxiolytic (bromazepam) and antidepressant drugs (fluoxetine and nortriptyline) on behavioral (novel tank test), biochemical (whole-body cortisol), and molecular parameters (cox-2, tnf-α, il-6, and il-10 gene expression) in zebrafish subjected to UCS. We replicated previous data showing that UCS induces behavioral and neuroendocrine alterations in zebrafish, and we show for the first time that anxiolytic and antidepressant drugs are able to prevent such effects. Furthermore, we extended the molecular characterization of the model, revealing that UCS increases expression of the pro-inflammatory markers cox-2 and il-6, which was also prevented by the drugs tested. This study reinforces the use of zebrafish as a model organism to study the behavioral and physiological effects of stress. The UCS protocol may also serve as a screening tool for evaluating new drugs that can be used to treat psychiatric disorders with stress-related etiologies.

Despite the recent advances in understanding the pathophysiology of anxiety disorders, the pharmacological treatments currently available are limited in efficacy and induce serious side effects. A possible strategy to achieve clinical benefits is drug repurposing, i.e., discovery of novel applications for old drugs, bringing new treatment options to the market and to the patients who need them. N-acetylcysteine (NAC), a commonly used mucolytic and paracetamol antidote, has emerged as a promising molecule for the treatment of several neuropsychiatric disorders. The mechanism of action of this drug is complex, and involves modulation of antioxidant, inflammatory, neurotrophic and glutamate pathways. Here we evaluated the effects of NAC on behavioral parameters relevant to anxiety in zebrafish. NAC did not alter behavioral parameters in the novel tank test, prevented the anxiety-like behaviors induced by an acute stressor (net chasing), and increased the time zebrafish spent in the lit side in the light/dark test. These data may indicate that NAC presents an anti-stress effect, with the potential to prevent stress-induced psychiatric disorders such as anxiety and depression. The considerable homology between mammalian and zebrafish genomes invests the current data with translational validity for the further clinical trials needed to substantiate the use of NAC in anxiety disorders.

Oxidative stress and DNA damage are involved in the glyphosate-based herbicide toxicity. Uncaria tomentosa (UT; Rubiaceae) is a plant species from South America containing bioactive compounds with known beneficial properties. The objective of this work was to evaluate the antioxidant and antigenotoxic potential of UT extract in a model of acute exposure to glyphosate-Roundup® (GR) in zebrafish (Danio rerio). We showed that UT (1.0 mg/mL) prevented the decrease of brain total thiols, the increase of lipid peroxidation in both brain and liver, and the decrease of liver GPx activity caused after 96 h of GR (5.0 mg/L) exposure. In addition, UT partially protected against the increase of micronucleus frequency induced by GR exposure in fish brain. Overall, our results indicate that UT protects against damage induced by a glyphosate-based herbicide by providing antioxidant and antigenotoxic effects, which may be related to the phenolic compounds identified in the extract.

Chronic stress may cause physical, behavioral and neuropsychiatric changes, affecting the health condition of an individual. Aggression is a universal behavior with great relevance on human and animal social systems. Despite studies showing the influence of chronic stress on aggression, the effects of unpredictable chronic stress (UCS) on aggressive behavior in male and female zebrafish remain unknown. Thus, the aim of this study was to evaluate the effects of UCS on the aggressive behavior and cortisol levels in adult zebrafish of both sexes. Our results showed that UCS increased aggression in males, but not in females, which displayed more aggressive behavior at baseline than control males. Increased whole-body cortisol levels were observed in stressed males; however, no differences were found between female groups. In conclusion, we reported for the first time gender differences on behavioral parameters and cortisol levels in response to UCS in zebrafish. These results highlight the relevance of studying behavioral and physiological parameters in both sexes separately.

Hydropower offers a reliable source of electricity in several countries, and Brazil supplies its energy needs almost entirely through hydropower plants. Nevertheless, hydropower plants comprise large buildings and water reservoirs and dams, resulting in huge ecological disruptions. Here, we analyzed the impact of four hydropower reservoirs construction in metals and pesticides incidence and the cytotoxic and genotoxic potential of sediment elutriate of rivers from southern Brazil. Our analyses have evidenced the elevated incidence of different metals (lead, iron, cadmium, and chrome) and pesticides (methyl parathion, atrazine, and 2,4-dichlorophenoxyacetic acid). We showed that Allium cepa exposed to sediment elutriates did not change the seed germination rate and mitotic index. However, roots from Allium cepa exposed to reservoirs sediment elutriates showed increased occurrence of chromosomal aberrations and nuclear abnormalities. Therefore, the results obtained in our study indicate that sediment from reservoirs present elevated concentration of metals and pesticides and a significant genotoxic potential. Taken together, our data support that hydropower reservoirs represent an environmental scenario that could impact surrounding wildlife and population.

This study evaluated water parameters in ponds affected by coal extraction. Allium cepa assay was used to measure genotoxicity/mutagenicity of the sediment. Samples were collected from four ponds in the southern state of Santa Catarina. Water temperature, pH, dissolved oxygen, conductivity and turbidity were measured. Sediments were analyzed for heavy metals. Elutriate samples were prepared at a ratio of 1:4 sediment:water. Allium cepa bulbs were placed in samples prepared from each pond, with ultrapure water used as negative control and methyl methane sulfonate as positive control. Root length, mitotic index, chromosomal aberrations, micronuclei, and nuclear abnormalities were measured. The pH of two ponds, as well as electrical conductivity and dissolved oxygen of all ponds were below the minimum limits set by Brazilian regulation. All heavy metals analyzed were found in all sediment samples, but only Cd concentration was above the legal limit set by Brazilian law. Allium cepa root growth for samples from Ponds 1, 2, and 4 was significantly lower than the negative control. Meristematic cells exposed to elutriate samples showed no significant changes in cell division. There was a significant increase in total chromosomal aberrations in all treated samples in comparison with the negative control. This study demonstrates that even low concentrations of heavy metals can damage exposed biota, possibly due to synergistic effects. We also found the A. cepa bioassay to be a simple and useful tool for genotoxicity/mutagenicity analyses, and recommend its use for environmental monitoring and management in areas influenced by mining activities. Resumo Este estudo analisou parâmetros de qualidade da água em lagoas sob influência de extração de carvão na região sul do estado de Santa Catarina. A genotoxicidade/mutagenicidade de sedimentos foram analisadas usando-se o ensaio Allium cepa. A temperatura, pH, oxigênio dissolvido, condutividade e turbidez foram mensuradas em sete lagoas, bem como, os metais pesados presentes nos sedimentos. Amostras de elutriato foram preparadas em proporção 1:4 (sedimento: água). Raízes de Allium cepa foram expostas aos elutriatos obtidos dos sedimentos coletados, sendo que raízes dos grupos controle negativo e positivo foram expostas à água ultrapura e metil-metano-sulfonato, respectivamente. Após a exposição, foram analisados: comprimento das raízes, índice mitótico, aberrações cromossômicas, micronúcleos e anormalidades nucleares. O pH de duas lagoas, e a condutividade e o nível de oxigênio dissolvido de todas as lagoas estavam abaixo do limite estabelecido pela legislação brasileira. Todas as amostras continham metais pesados e o Cd encontrava-se em nível superior ao permitido pela legislação. Raízes expostas a sedimentos das lagoas 1, 2 e 4 tiveram diminuição de crescimento. Raízes expostas ao elutriato de todas as lagoas não apresentaram alterações na divisão celular. Houve aumento significativo no número de aberrações cromossômicas em raízes expostas em todas as amostras das lagoas em comparação com ao controle negativo. Este estudo mostra que, mesmo em baixas concentrações, metais pesados podem danificar a biota, possivelmente devido a efeitos sinérgicos. Pode ser indicado que o ensaio em Allium cepa é uma ferramenta simples e útil para análises de genotoxicidade/mutagenicidade, assim como pode ser recomendado seu uso em projetos de monitoramento e gestão ambiental de áreas influenciadas por atividades de mineração.

Animal behaviour is the interaction between environment and an individual organism, which also can be influenced by its neighbours. Variations in environmental conditions, as those caused by contaminants, may lead to neurochemical impairments altering the pattern of the behavioural repertoire of the species. Atrazine (ATZ) is an herbicide widely used in agriculture that is frequently detected in surface water, affecting non-target species. The zebrafish is a valuable model organism to assess behavioural and neurochemical effects of different contaminants since it presents a robust behavioural repertoire and also all major neurotransmitter systems described for mammalian species. The goal of this study was to evaluate the effects of subchronic ATZ exposure in defensive behaviours of zebrafish (shoaling, thigmotaxis, and depth preference) using the split depth tank. Furthermore, to investigate a putative role of cholinergic signalling on ATZ-mediated effects, we tested whether this herbicide alters acetylcholinesterase (AChE) activity in brain and muscle preparations. Fish were exposed to ATZ for 14 days and the following groups were tested: control (0.2% acetone) and ATZ (10 and 1000 μg/L). The behaviour of four animals in the same tank was recorded for 6 min and biological samples were prepared. Our results showed that 1000 μg/L ATZ significantly increased the inter-fish distance, as well as the nearest and farthest neighbour distances. This group also presented an increase in the shoal area with decreased social interaction. No significant differences were detected for the number of animals in the shallow area, latency to enter the shallow and time spent in shallow and deep areas of the apparatus, but the ATZ 1000 group spent significantly more time near the walls. Although ATZ did not affect muscular AChE, it significantly reduced AChE activity in brain. Exposure to 10 μg/L ATZ did not affect behaviour or AChE activity. These data suggest that ATZ impairs defensive behaviours of zebrafish, which could be related to its action on brain cholinergic neurotransmission. Moreover, the use of the split depth tank could be an alternative strategy to assess group behaviour and depth preference after exposure to chemical compounds.