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1.
Aquat Toxicol ; 234: 105798, 2021 May.
Artigo em Inglês | MEDLINE | ID: mdl-33799113

RESUMO

A wide variety of active pharmaceutical ingredients are released into the environment and pose a threat to aquatic organisms. Drug products using micro- and nanoparticle technology can lower these emissions into the environment by their increased bioavailability to the human patients. However, due to this enhanced efficacy, micro- and nanoscale drug delivery systems can potentially display an even higher toxicity, and thus also pose a risk to non-target organisms. Fenofibrate is a lipid-regulating agent and exhibits species-related hazards in fish. The ecotoxic effects of a fenofibrate formulation embedded into a hydroxypropyl methylcellulose microparticle matrix, as well as those of the excipients used in the formulation process, were evaluated. To compare the effects of fenofibrate without a formulation, fenofibrate was dispersed in diluted ISO water alone or dissolved in the solvent DMF and then added to diluted ISO water. The effects of these various treatments were assessed using the fish embryo toxicity test, acridine orange staining and gene expression analysis assessed by quantitative RT polymerase chain reaction. Exposure concentrations were assessed by chemical analysis. The effect threshold concentrations of fenofibrate microparticle precipitates were higher compared to the formulation. Fenofibrate dispersed in 20%-ISO-water displayed the lowest toxicity. For the fenofibrate formulation as well as for fenofibrate added as a DMF solution, greater ecotoxic effects were observed in the zebrafish embryos. The chemical analysis of the solutions revealed that more fenofibrate was present in the samples with the fenofibrate formulation as well as fenofibrate added as a DMF solution compared to fenofibrate dispersed in diluted ISO water. This could explain the higher ecotoxicity. The toxic effects on the zebrafish embryo thus suggested that the formulation as well as the solvent increased the bioavailability of fenofibrate.


Assuntos
Fenofibrato/toxicidade , Poluentes Químicos da Água/toxicidade , Peixe-Zebra/crescimento & desenvolvimento , Animais , Cromatografia Líquida de Alta Pressão , Composição de Medicamentos , Embrião não Mamífero/efeitos dos fármacos , Embrião não Mamífero/metabolismo , Fenofibrato/análise , Fenofibrato/química , Regulação da Expressão Gênica/efeitos dos fármacos , Espectrometria de Massas , Tamanho da Partícula , Testes de Toxicidade , Peixe-Zebra/metabolismo
2.
Aquat Toxicol ; 234: 105813, 2021 May.
Artigo em Inglês | MEDLINE | ID: mdl-33812311

RESUMO

The production of large amounts of synthetic industrial and biomedical compounds, together with environmental pollutants, poses a risk to our ecosystem and induces negative effects on the health of wildlife and human beings. With the emergence of the global problem of chemical contamination, the adverse biological effects of these chemicals are gaining attention among the scientific communities, industry, governments, and the public. Among these chemicals, endocrine disrupting chemicals (EDCs) are regarded as one of the major global issues that potentially affecting our health. There is an urgent need of understanding the potential hazards of such chemicals. Zebrafish have been widely used in the aquatic toxicology. In this review, we first discuss the strategy of transgenic lines that used in the toxicological studies, followed by summarizing the current omics approaches (transcriptomics, proteomics, metabolomics, and epigenomics) on toxicities of EDCs in this model. We will also discuss the possible transgenerational effects in zebrafish and future prospective of the integrated omics approaches with customized transgenic organism. To conclude, we summarize the current findings in the field, and provide our opinions on future environmental toxicity research in the zebrafish model.


Assuntos
Epigenômica , Metabolômica , Proteômica , Poluentes Químicos da Água/toxicidade , Peixe-Zebra/metabolismo , Animais , Animais Geneticamente Modificados/metabolismo , Ecossistema , Disruptores Endócrinos/toxicidade , Modelos Animais , Peixe-Zebra/crescimento & desenvolvimento
3.
Aquat Toxicol ; 233: 105794, 2021 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-33662880

RESUMO

Polychlorinated biphenyls (PCBs) and 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) are environmental contaminants known to impact cardiac development, a key step in the embryonic development of most animals. To date, little is understood of the molecular mechanism driving the observed cardiac defects in exposed fishes. The literature shows PCB & TCDD derived cardiac defects are concurrent with, but not caused by, expression of cyp1A, due to activation of the aryl hydrocarbon receptor (AhR) gene activation pathway. However, in this study, detailed visualization of fish hearts exposed to PCBs and TCDD show that, in addition to a failure of cardiac looping in early heart development, the inner endocardial lining of the heart fails to maintain proper cell adhesion and tissue integrity. The resulting gap between the endocardium and myocardium in both zebrafish and Atlantic sturgeon suggested functional faults in endothelial adherens junction formation. Thus, we explored the molecular mechanism triggering cardiac defects using immunohistochemistry to identify the location and phosphorylation state of key regulatory and adhesion molecules. We hypothesized that PCB and TCDD activates AhR, phosphorylating Src, which then phosphorylates the endothelial adherens junction protein, VEcadherin. When phosphorylated, VEcadherin dimers, found in the endocardium and vasculature, separate, reducing tissue integrity. In zebrafish, treatment with PCB and TCDD contaminants leads to higher phosphorylation of VEcadherin in cardiac tissue suggesting that these cells have reduced connectivity. Small molecule inhibition of Src phosphorylation prevents contaminant stimulated phosphorylation of VEcadherin and rescues both cardiac function and gross morphology. Atlantic sturgeon hearts show parallels to contaminant exposed zebrafish cardiac phenotype at the tissue level. These data suggest that the mechanism for PCB and TCDD action in the heart is, in part, distinct from the canonical mechanism described in the literature and that cardiac defects are impacted by this nongenomic mechanism.


Assuntos
Embrião não Mamífero/efeitos dos fármacos , Coração/efeitos dos fármacos , Bifenilos Policlorados/toxicidade , Dibenzodioxinas Policloradas/toxicidade , Receptores de Hidrocarboneto Arílico/metabolismo , Poluentes Químicos da Água/toxicidade , Peixe-Zebra/metabolismo , Animais , Sinergismo Farmacológico , Embrião não Mamífero/anormalidades , Embrião não Mamífero/metabolismo , Desenvolvimento Embrionário/efeitos dos fármacos , Coração/embriologia , Miocárdio/metabolismo , Peixe-Zebra/crescimento & desenvolvimento
4.
J Vis Exp ; (168)2021 02 09.
Artigo em Inglês | MEDLINE | ID: mdl-33645566

RESUMO

Reactive oxygen species (ROS) are well-established signaling molecules, which are important in normal development, homeostasis, and physiology. Among the different ROS, hydrogen peroxide (H2O2) is best characterized with respect to roles in cellular signaling. H2O2 has been implicated during the development in several species. For example, a transient increase in H2O2 has been detected in zebrafish embryos during the first days following fertilization. Furthermore, depleting an important cellular H2O2 source, NADPH oxidase (NOX), impairs nervous system development such as the differentiation, axonal growth, and guidance of retinal ganglion cells (RGCs) both in vivo and in vitro. Here, we describe a method for imaging intracellular H2O2 levels in cultured zebrafish neurons and whole larvae during development using the genetically encoded H2O2-specific biosensor, roGFP2-Orp1. This probe can be transiently or stably expressed in zebrafish larvae. Furthermore, the ratiometric readout diminishes the probability of detecting artifacts due to differential gene expression or volume effects. First, we demonstrate how to isolate and culture RGCs derived from zebrafish embryos that transiently express roGFP2-Orp1. Then, we use whole larvae to monitor H2O2 levels at the tissue level. The sensor has been validated by the addition of H2O2. Additionally, this methodology could be used to measure H2O2 levels in specific cell types and tissues by generating transgenic animals with tissue-specific biosensor expression. As zebrafish facilitate genetic and developmental manipulations, the approach demonstrated here could serve as a pipeline to test the role of H2O2 during neuronal and general embryonic development in vertebrates.


Assuntos
Técnicas Biossensoriais/métodos , Peróxido de Hidrogênio/metabolismo , Imagem Molecular/métodos , Neurogênese , Espécies Reativas de Oxigênio/metabolismo , Células Ganglionares da Retina/metabolismo , Peixe-Zebra/metabolismo , Animais , Células Cultivadas , Peróxido de Hidrogênio/análise , Oxirredução , Células Ganglionares da Retina/citologia , Peixe-Zebra/crescimento & desenvolvimento
5.
Int J Mol Sci ; 22(4)2021 Feb 11.
Artigo em Inglês | MEDLINE | ID: mdl-33670431

RESUMO

A reduction in daily caloric or nutrient intake has been observed to promote health benefits in mammals and other vertebrates. Feed Restriction (FR), whereby the overall food intake of the organism is reduced, has been explored as a method to improve metabolic and immune health, as well as to optimize productivity in farming. However, less is known regarding the molecular and physiological consequences of FR. Using the model organism, Danio rerio, we investigated the impact of a short-term (month-long) FR on growth, gut morphology and gene expression. Our data suggest that FR has minimal effects on the average growth rates, but it may affect weight and size heterogeneity in a sex-dependent manner. In the gut, we observed a significant reduction in gut circumference and generally lower mucosal heights, whereas other parameters remained unchanged. Gene Ontology (GO), EuKaryotic Orthologous Groups (KOG), and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis identified numerous metabolic, reproductive, and immune response pathways that were affected by FR. These results broaden our understanding of FR and contribute towards growing knowledge of its effects on vertebrate health.


Assuntos
Ingestão de Alimentos , Ingestão de Energia , Regulação da Expressão Gênica , Intestinos/crescimento & desenvolvimento , Caracteres Sexuais , Peixe-Zebra/crescimento & desenvolvimento , Ração Animal , Animais , Feminino , Intestinos/anatomia & histologia , Masculino , Peixe-Zebra/anatomia & histologia
6.
Carbohydr Polym ; 259: 117749, 2021 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-33673979

RESUMO

The increasing world-wide demand for food has prompted the development of efficient and environmentally friendly pesticide formulations. In this article, we have prepared CMC-g-PRSG carrier based on two compounds from natural materials carboxymethyl cellulose (CMC) and rosin (RS). The model pesticide avermectin (AVM) was encapsulated through hydrophobic interaction, and self-assembled to form nanopesticide AVM@CMC-g-PRSG with an average particle size of 167 nm. The prepared nanopesticide displays enhanced dispersibility and stability of AVM in water, and can effectively adhere to the leaves to prevent loss. The release rate of AVM encapsulated in the nanocarrier can be controlled by adjusting pH, and AVM half-life under ultraviolet radiation shows a 3-fold increase allowing control of pests for prolonged periods of time in practical applications. Biological safety tests showed that AVM@CMC-g-PRSG effectively reduces the toxicity of AVM to aquatic animals. Therefore, the cheap and degradable carrier CMC-g-PRSG can improve the effect of hydrophobic pesticides.


Assuntos
Carboximetilcelulose Sódica/química , Portadores de Fármacos/química , Resinas Vegetais/química , Animais , Liberação Controlada de Fármacos , Meia-Vida , Concentração de Íons de Hidrogênio , Interações Hidrofóbicas e Hidrofílicas , Ivermectina/análogos & derivados , Ivermectina/química , Ivermectina/metabolismo , Ivermectina/farmacologia , Larva/efeitos dos fármacos , Nanopartículas/química , Tamanho da Partícula , Controle de Pragas/métodos , Praguicidas/química , Praguicidas/metabolismo , Praguicidas/farmacologia , Raios Ultravioleta , Peixe-Zebra/crescimento & desenvolvimento , Peixe-Zebra/fisiologia
7.
Carbohydr Polym ; 259: 117710, 2021 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-33673989

RESUMO

Harmful algal blooms induce severe environmental problems. It is challenging to remove algae by the current available treatments involving complicate process and costly instruments. Here, we developed a CaO2@PEG-loaded water-soluble self-branched chitosan (CP-SBC) system, which can remove algae from water in one-step without additional instrumentation. This approach utilizes a novel flocculant (self-branched chitosan) integrated with flotation function (induced by CaO2@PEG). CP-SBC exhibited better flocculation performance than commercial flocculants, which is attributed to the enhanced bridging and sweeping effect of branched chitosan. CP-SBC demonstrated outstanding biocompatibility, which was verified by zebrafish test and algae activity test. CaO2@PEG-loaded self-branched chitosan can serve as an "Air flotation system" to spontaneous float the flocs after flocculation by sustainably released O2. Furthermore, CP-SBC can improve water quality through minimizing dissolved oxygen depletion and reducing total phosphorus concentrations.


Assuntos
Quitosana/química , Proliferação Nociva de Algas/fisiologia , Animais , Materiais Biocompatíveis/química , Materiais Biocompatíveis/farmacologia , Compostos de Cálcio/química , Floculação/efeitos dos fármacos , Proliferação Nociva de Algas/efeitos dos fármacos , Cinética , Larva/efeitos dos fármacos , Óxidos/química , Oxigênio/química , Fósforo/química , Polietilenoglicóis/química , Porosidade , Peixe-Zebra/crescimento & desenvolvimento , Peixe-Zebra/fisiologia
8.
J Vis Exp ; (168)2021 02 10.
Artigo em Inglês | MEDLINE | ID: mdl-33645565

RESUMO

Understanding the ephemeral changes that occur during brain development and maturation requires detailed high-resolution imaging in space and time at cellular and subcellular resolution. Advances in molecular and imaging technologies have allowed us to gain numerous detailed insights into cellular and molecular mechanisms of brain development in the transparent zebrafish embryo. Recently, processes of refinement of neuronal connectivity that occur at later larval stages several weeks after fertilization, which are for example control of social behavior, decision making or motivation-driven behavior, have moved into focus of research. At these stages, pigmentation of the zebrafish skin interferes with light penetration into brain tissue, and solutions for embryonic stages, e.g., pharmacological inhibition of pigmentation, are not feasible anymore. Therefore, a minimally invasive surgical solution for microscopy access to the brain of awake zebrafish is provided that is derived from electrophysiological approaches. In teleosts, skin and soft skull cartilage can be carefully removed by micro-peeling these layers, exposing underlying neurons and axonal tracts without damage. This allows for recording neuronal morphology, including synaptic structures and their molecular contents, and the observation of physiological changes such as Ca2+ transients or intracellular transport events. In addition, interrogation of these processes by means of pharmacological inhibition or optogenetic manipulation is feasible. This brain exposure approach provides information about structural and physiological changes in neurons as well as the correlation and interdependence of these events in live brain tissue in the range of minutes or hours. The technique is suitable for in vivo brain imaging of zebrafish larvae up to 30 days post fertilization, the latest developmental stage tested so far. It, thus, provides access to such important questions as synaptic refinement and scaling, axonal and dendritic transport, synaptic targeting of cytoskeletal cargo or local activity-dependent expression. Therefore, a broad use for this mounting and imaging approach can be anticipated.


Assuntos
Encéfalo/diagnóstico por imagem , Encéfalo/crescimento & desenvolvimento , Imageamento Tridimensional , Pele/diagnóstico por imagem , Crânio/diagnóstico por imagem , Peixe-Zebra/crescimento & desenvolvimento , Anestesia , Animais , Encéfalo/irrigação sanguínea , Larva/fisiologia , Neurônios/fisiologia , Vigília/fisiologia
9.
Aquat Toxicol ; 234: 105786, 2021 May.
Artigo em Inglês | MEDLINE | ID: mdl-33735685

RESUMO

The aryl hydrocarbon receptor (AHR) has endogenous functions in mammalian vascular development and is necessary for mediating the toxic effects of a number of environmental contaminants. Studies in mice have demonstrated that AHR is necessary for the formation of the renal, retinal, and hepatic vasculature. In fish, exposure to the prototypic AHR agonist 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) induces expression of the AHR biomarker cyp1a throughout the developing vasculature and produces vascular malformations in the head and heart. However, it is not known whether the vascular structures that are sensitive to loss of AHR function are also disrupted by aberrant AHR activation. Here, we report that TCDD-exposure in zebrafish disrupts development of 1) the subintestinal venous plexus (SIVP), which vascularizes the developing liver, kidney, gut, and pancreas, and 2) the superficial annular vessel (SAV), an essential component of the retinal vasculature. Furthermore, we determined that TCDD exposure increased the expression of bmp4, a key molecular mediator of SIVP morphogenesis. We hypothesize that the observed SIVP phenotypes contribute to one of the hallmarks of TCDD exposure in fish - the failure of the yolk sac to absorb. Together, our data describe novel TCDD-induced vascular phenotypes and provide molecular insight into critical factors producing the observed vascular malformations.


Assuntos
Dibenzodioxinas Policloradas/toxicidade , Veia Retiniana/efeitos dos fármacos , Poluentes Químicos da Água/toxicidade , Peixe-Zebra/metabolismo , Animais , Animais Geneticamente Modificados/metabolismo , Proteína Morfogenética Óssea 4/genética , Proteína Morfogenética Óssea 4/metabolismo , Embrião não Mamífero/efeitos dos fármacos , Embrião não Mamífero/metabolismo , Fígado/irrigação sanguínea , Veia Retiniana/crescimento & desenvolvimento , Veias/efeitos dos fármacos , Peixe-Zebra/crescimento & desenvolvimento , Proteínas de Peixe-Zebra/genética , Proteínas de Peixe-Zebra/metabolismo
10.
Eur J Med Chem ; 216: 113307, 2021 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-33652354

RESUMO

Androgen receptor (AR) is an effective therapeutic target for the treatment of prostate cancer. We report herein the design, synthesis, and biological evaluation of highly effective proteolysis targeting chimeras (PROTAC) androgen receptor (AR) degraders, such as compound A031. It could induce the degradation of AR protein in VCaP cell lines in a time-dependent manner, achieving the IC 50 value of less than 0.25 µM. The A031 is 5 times less toxic than EZLA and works with an appropriate half-life (t 1/2) or clearance rate (Cl). Also, it has a significant inhibitory effect on tumor growth in zebrafish transplanted with human prostate cancer (VCaP). Therefore, A031 provides a further idea of developing novel drugs for prostate cancer.


Assuntos
Antagonistas de Receptores de Andrógenos/química , Receptores Androgênicos/metabolismo , Antagonistas de Receptores de Andrógenos/farmacocinética , Antagonistas de Receptores de Andrógenos/farmacologia , Antagonistas de Receptores de Andrógenos/uso terapêutico , Animais , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Desenho de Fármacos , Avaliação Pré-Clínica de Medicamentos , Meia-Vida , Humanos , Masculino , Neoplasias da Próstata/tratamento farmacológico , Neoplasias da Próstata/mortalidade , Neoplasias da Próstata/patologia , Proteólise , Ratos , Ratos Sprague-Dawley , Receptores Androgênicos/genética , Relação Estrutura-Atividade , Taxa de Sobrevida , Transplante Heterólogo , Peixe-Zebra/crescimento & desenvolvimento , Peixe-Zebra/fisiologia
11.
Eur J Med Chem ; 216: 113262, 2021 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-33711764

RESUMO

The number of cancer cases continues to increase worldwide, and unfortunately the main systemic treatments available have numerous of side effects. Ruthenium complexes have shown to be promising chemotherapeutic agents, since they present low toxicity and are more selective for tumor tissues. We report the synthesis, characterization and biological properties of two new ruthenium (II) complexes containing Lapachol and Lawsone as ligands: (1) [Ru(Law)(dppb)(phen)]PF6 and (2) [Ru(Lap)(dppb)(phen)]PF6, where Law = Lawsone, Lap = Lapachol, dppb = 1,4-bis(diphenylphosphine)butane and phen = 1,10-phenanthroline. The ability of the complexes (1) and (2) to interact with CT-DNA (Calf Thymus) was investigated, and the results indicate that the complexes have shown a weak interaction with this macromolecule. Complexes (1) and (2) showed a moderate interaction with BSA, via a spontaneous process with the involvement of van der Waals and hydrogen bond interactions. Both complexes were tested against human lung cancer cell lines, chronic human myeloid leukemia, murine melanoma and human cervical and non-tumoral murine fibroblast adenocarcinoma, human lung fibroblasts and monkey kidney epithelia. The potential for cytotoxicity was tested out using the MTT assay and the neutral red test, to calculate inhibitory concentrations (IC50) and selectivity indices (IS). Both complexes showed a higher selectivity index of 1.17 and 10.91, respectively, for the HeLa tumor line. Studies of toxicological evaluation, using the micronucleus test and the comet assay against non-tumor cells, as well as an assessment of the potential for acute toxicity and neurotoxicity in zebrafish (Danio rerio). In the in vitro micronucleus test, complex (1) showed the least genotoxic potential, and in the in vitro comet assay both compounds had revealed a genotoxic potential at 0.5 and 1.0 mg L-1, with no difference between 24 h and 48 h exposure times. In the acute toxicity tests on zebrafish embryos, complex (1) showed sublethal effects such as decreased blood circulation and heartbeat rate, which were less pronounced than with complex (2). In contrast to complex 2, which caused lethality even before 48h, complex (1) did not cause the death of the embryos at concentrations up to (2.0 mg L-1). Complex (2) also lead to a delay in the embryo. Cell based in vitro methods thus proved able to provide specific toxicological data, allowing a significant reduction in ∖animal experimentation. Given that in vitro tests cannot completely replace animal tests, the use of less advanced developmental stages such as zebrafish embryos, which - at least in the European Union - are not regarded protected, could be shown to be an excellent alternative for testing with, e.g., mammals.


Assuntos
Complexos de Coordenação/química , Rutênio/química , Animais , Antineoplásicos/química , Antineoplásicos/metabolismo , Antineoplásicos/farmacologia , Bovinos , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Complexos de Coordenação/metabolismo , Complexos de Coordenação/farmacologia , Cristalografia por Raios X , DNA/química , DNA/metabolismo , Embrião não Mamífero/efeitos dos fármacos , Embrião não Mamífero/metabolismo , Humanos , Conformação Molecular , Ligação Proteica , Soroalbumina Bovina/química , Soroalbumina Bovina/metabolismo , Termodinâmica , Peixe-Zebra/crescimento & desenvolvimento
12.
Ecotoxicol Environ Saf ; 214: 112107, 2021 May.
Artigo em Inglês | MEDLINE | ID: mdl-33667734

RESUMO

Perfluorobutanesulfonate (PFBS) pollutant and probiotic bacteria can interact to affect the reproductive outcomes of zebrafish. However, it is still unexplored how the growth and health of offspring are modulated by the combination of PFBS and probiotic. In the present study, adult zebrafish were exposed to 0 and 10 µg/L PFBS for 40 days, with or without dietary supplementation of probiotic Lactobacillus rhamnosus. After parental exposure, the development, growth and viability of offspring larvae were examined, with the integration of molecular clues across proteome fingerprint, growth hormone/insulin-like growth factor (GH/IGF) axis, calcium homeostasis, hypothalamic-pituitary-adrenal (HPA) axis and nutrient metabolism. Parental probiotic supplementation significantly increased the body weight and body length of offspring larvae. Despite the spiking of PFBS, larvae from the combined exposure group still had longer body length. RNA processing and ribosomal assembly pathways may underlie the enhancement of offspring growth by probiotic bacteria. However, the presence of PFBS remarkably increased the concentrations of cortisol hormone in offspring larvae as means to cope with the xenobiotic stress, which required more energy production. As evidenced by the proteomic analysis, the addition of probiotic bacteria likely alleviated the energy metabolism disorders of PFBS, thus allocating more energy for the larval offspring growth from the combined group. It was noteworthy that multiple molecular disturbances caused by PFBS were antagonized by probiotic additive. Overall, the present study elucidated the intergenerational interaction between PFBS and probiotic on offspring growth and health after parental exposure.


Assuntos
Fluorcarbonetos/toxicidade , Lactobacillus rhamnosus , Larva/efeitos dos fármacos , Probióticos/farmacologia , Ácidos Sulfônicos/toxicidade , Poluentes Químicos da Água/toxicidade , Peixe-Zebra/crescimento & desenvolvimento , Animais , Metabolismo Energético/efeitos dos fármacos , Feminino , Sistema Hipotálamo-Hipofisário , Larva/crescimento & desenvolvimento , Masculino , Proteômica
13.
Int J Mol Sci ; 22(4)2021 Feb 07.
Artigo em Inglês | MEDLINE | ID: mdl-33562361

RESUMO

Natural mycelial exopolysaccharide (EPS) and endopolysaccharide (ENS) extracted from bioreactor-cultivated European Ganoderma applanatum mushrooms are of potential high commercial value for both food and adjacent biopharmaceutical industries. In order to evaluate their potential toxicity for aquaculture application, both EPS (0.01-10 mg/mL) and ENS (0.01-10 mg/mL) extracts were tested for Zebrafish Embryo Toxicity (ZFET); early development effects on Zebrafish Embryos (ZE) were also analyzed between 24 and 120 h post-fertilization (HPF). Both EPS and ENS are considered non-toxic with LC50 of 1.41 mg/mL and 0.87 mg/mL respectively. Both EPS and ENS did not delay hatching and teratogenic defect towards ZE with <1.0 mg/mL, respectively. No significant changes in the ZE heart rate were detected following treatment with the two compounds tested (EPS: 0.01-10 mg/mL: 176.44 ± 0.77 beats/min and ENS: 0.01-10 mg/mL: 148.44 ± 17.75 beats/min) compared to normal ZE (120-180 beats/min). These initial findings support future pre-clinical trials in adult fish models with view to safely using EPS and ENS as potential feed supplements for supplements for development of the aquaculture industry.


Assuntos
Reatores Biológicos/microbiologia , Embrião não Mamífero/citologia , Ganoderma/química , Micélio/química , Polissacarídeos/toxicidade , Testes de Toxicidade/métodos , Peixe-Zebra/crescimento & desenvolvimento , Animais , Bioensaio , Embrião não Mamífero/efeitos dos fármacos , Europa (Continente)
14.
PLoS One ; 16(1): e0244151, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33417603

RESUMO

Machine learning approaches are becoming increasingly widespread and are now present in most areas of research. Their recent surge can be explained in part due to our ability to generate and store enormous amounts of data with which to train these models. The requirement for large training sets is also responsible for limiting further potential applications of machine learning, particularly in fields where data tend to be scarce such as developmental biology. However, recent research seems to indicate that machine learning and Big Data can sometimes be decoupled to train models with modest amounts of data. In this work we set out to train a CNN-based classifier to stage zebrafish tail buds at four different stages of development using small information-rich data sets. Our results show that two and three dimensional convolutional neural networks can be trained to stage developing zebrafish tail buds based on both morphological and gene expression confocal microscopy images, achieving in each case up to 100% test accuracy scores. Importantly, we show that high accuracy can be achieved with data set sizes of under 100 images, much smaller than the typical training set size for a convolutional neural net. Furthermore, our classifier shows that it is possible to stage isolated embryonic structures without the need to refer to classic developmental landmarks in the whole embryo, which will be particularly useful to stage 3D culture in vitro systems such as organoids. We hope that this work will provide a proof of principle that will help dispel the myth that large data set sizes are always required to train CNNs, and encourage researchers in fields where data are scarce to also apply ML approaches.


Assuntos
Aprendizado Profundo , Embrião não Mamífero/metabolismo , Peixe-Zebra/metabolismo , Animais , Embrião não Mamífero/patologia , Expressão Gênica , Processamento de Imagem Assistida por Computador , Microscopia Confocal , Cauda/metabolismo , Cauda/patologia , Peixe-Zebra/crescimento & desenvolvimento
15.
Nanotoxicology ; 15(2): 257-275, 2021 03.
Artigo em Inglês | MEDLINE | ID: mdl-33503388

RESUMO

Safety on the use of magnetic nanomaterials (MNMs) has become an active topic of research given all the recent applications of these materials in various fields. It is known that the toxicity of MNMs depends on size, shape, and surface functionalization. In this study, we evaluate the biocompatibility with different aquatic organisms of engineered MNMs-CIT with excellent aqueous dispersion and long-term colloidal stability. Primary producers (the alga Pseudokirchneriella subcapitata), primary consumers (the rotifer Lecane papuana), and predators (the fish, Danio rerio) interacted with these materials in acute and sub-chronic toxicity tests. Our results indicate that P. subcaptita was the most sensitive taxon to MNMs-CIT. Inhibition of their population growth (IC50 = 22.84 mg L-1) elicited cell malformations and increased the content of photosynthetic pigments, likely due to inhibition of cell division (as demonstrated in AFM analysis). For L. papuana, the acute exposure to MNMs shows no significant mortality. However, adverse effects such as decreased rate of population and altered swimming patterns arise after chronic interaction with MNMs. For D. rerio organisms on early life stages, their exposure to MNMs results in delayed hatching of eggs, diminished survival of larvae, altered energy resources allocation (measured as the content of total carbohydrates, lipids, and protein), and increased glucose demand. As to our knowledge, this is the first study that includes three different trophic levels to assess the effect of MNMs in aquatic organisms; furthermore, we demonstrated that these MNMs pose hazards on aquatic food webs at low concentrations (few mgL-1).


Assuntos
Organismos Aquáticos/efeitos dos fármacos , Cloretos/toxicidade , Compostos Férricos/toxicidade , Compostos Ferrosos/toxicidade , Nanoestruturas/toxicidade , Poluentes Químicos da Água/toxicidade , Animais , Biomarcadores/análise , Cloretos/química , Compostos Férricos/química , Compostos Ferrosos/química , Cadeia Alimentar , Larva/efeitos dos fármacos , Fenômenos Magnéticos , Microalgas/efeitos dos fármacos , Nanoestruturas/química , Tamanho da Partícula , Rotíferos/efeitos dos fármacos , Propriedades de Superfície , Poluentes Químicos da Água/química , Peixe-Zebra/crescimento & desenvolvimento
16.
Ecotoxicol Environ Saf ; 208: 111585, 2021 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-33396108

RESUMO

Uranium is a radioactive element that is widely present in aquatic environment. However, limited knowledge is available about the effect of uranium on thyroid system, which plays a key role in the development of animals. In this study, zebrafish embryos were exposed to different environmentally relevant concentrations of uranium (2, 20 and 100 µg/L) for 120 h. The bioaccumulation, developmental toxicities, changes of thyroid hormones (THs) and key genes related to the hypothalamic-pituitary-thyroid (HPT) axis in larvae were analyzed after exposure. Results showed that uranium could bioaccumulate in zebrafish larvae, with the bioconcentration factors ranging from 49.6 to 523. Consequently, significant developmental toxicities and changes in locomotor activities were observed with a concentration-dependent manner. The levels of triiodothyronine (T3) levels in larvae were substantially decreased, whereas those of thyroxine (T4) were increased in fish bodies. The levels of THs were regulated by the negative feedback loops through HPT axis related genes, most of which (NIS, Deio1, Deio2, TRα, TSHß and UGT1ab) were significantly depressed after exposure to uranium. Our results suggest the potential toxicities and thyroid disruption of uranium on zebrafish, which would provide baseline data set for better understanding the impact of waterborne uranium on aquatic organisms and the associated mechanisms. This study also highlights the key role of thyroid disruption in the ecological risk assessment of uranium pollution.


Assuntos
Glândula Tireoide/efeitos dos fármacos , Hormônios Tireóideos/metabolismo , Urânio/toxicidade , Poluentes Radioativos da Água/toxicidade , Peixe-Zebra/fisiologia , Animais , Larva , Tiroxina , Tri-Iodotironina , Peixe-Zebra/crescimento & desenvolvimento , Proteínas de Peixe-Zebra/genética
17.
Ecotoxicol Environ Saf ; 208: 111700, 2021 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-33396031

RESUMO

Sertraline (SER) is one of the most frequently detected antidepressant drugs in aquatic environments. However, knowledge regarding SER-induced behavioral alterations in fish is insufficient, as well as the mechanisms underlying SER-induced toxicity. The present study aimed to determine behavioral and molecular responses in larval fish following SER exposure with a focus on its mode of action. Zebrafish embryos (~6 h-post-fertilization, hpf) were exposed to one of three concentrations of SER (1, 10, 100 µg/L) for 6 days, respectively. Evaluated parameters included development, behavior, transcripts related to serotonin signaling, serotonin levels, and acetylcholinesterase activity. Accelerated hatching of zebrafish embryos was observed for those fish exposed to 100 µg/L SER at 54 hpf. Locomotor activity (e.g. distance moved and mobile cumulative duration) was significantly reduced in larval zebrafish following exposure to 10 and 100 µg/L SER. Conversely, larval fish showed increased dark-avoidance after exposure to 1-100 µg/L SER. Of the measured transcripts related to serotonin signaling, only serotonin transporter (serta) and serotonin receptor 2c (5-ht2c) mRNA levels were increased in fish in response to 10 µg/L SER treatment. However, serotonin levels were unaltered in larvae exposed to SER. There were no differences among groups in acetylcholinesterase activity at any concentration tested. Taking together, the results evidenced that exposure to SER alters behavioral responses in early-staged zebrafish, which may be related to the abnormal expression of 5-ht2c. This study elucidates molecular responses to SER and characterizes targets that may be sensitive to antidepressant pharmaceuticals in larval fish.


Assuntos
Antidepressivos/toxicidade , Comportamento Animal/efeitos dos fármacos , Sertralina/toxicidade , Poluentes Químicos da Água/toxicidade , Peixe-Zebra/fisiologia , Animais , Antidepressivos/análise , Embrião não Mamífero/efeitos dos fármacos , Embrião não Mamífero/fisiologia , Locomoção/efeitos dos fármacos , Serotonina/metabolismo , Sertralina/análise , Transdução de Sinais/efeitos dos fármacos , Poluentes Químicos da Água/análise , Peixe-Zebra/crescimento & desenvolvimento , Peixe-Zebra/metabolismo
18.
Ecotoxicol Environ Saf ; 208: 111613, 2021 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-33396133

RESUMO

The environmental effects of additives have attracted increasing attention. Sodium dehydroacetate (DHA-S), as an approved preservative, is widely added in processed foods, cosmetics and personal care products. However, DHA-S has been recently reported to induce hemorrhage and coagulation aberration in rats. Yet little is known about the ecotoxicological effect and underlying mechanisms of DHA-S. Here, we utilized the advantage of zebrafish model to evaluate such effects. DHA-S induced cerebral hemorrhage, mandibular dysplasia and pericardial edema in zebrafish after 24 h exposure (48-72 hpf) at 50 mg/L. We also observed the defective heart looping and apoptosis in DHA-S-treated zebrafish through o-dianisidine and acridine orange staining. Meanwhile, DHA-S induced the deficiency of Ca2+ and vitamin D3 in zebrafish. We further demonstrated that DHA-S stimulated Ca2+ influx resulting in Ca2+-dependent mitochondrial damage in cardiomyocytes. Additionally, DHA-S inhibited glucose uptake and repressed the biosynthesis of amino acids. Finally, we identified that sodium bicarbonate could rescue zebrafish from DHA-S induced cardiovascular toxicity. Altogether, our results suggest that DHA-S is a potential risk for cardiovascular system.


Assuntos
Cálcio/metabolismo , Embrião não Mamífero/efeitos dos fármacos , Desenvolvimento Embrionário/efeitos dos fármacos , Coração/efeitos dos fármacos , Pironas/toxicidade , Peixe-Zebra , Animais , Apoptose/efeitos dos fármacos , Cardiotoxicidade , Linhagem Celular , Hemorragia Cerebral/induzido quimicamente , Relação Dose-Resposta a Droga , Edema Cardíaco/induzido quimicamente , Coração/embriologia , Miocárdio/metabolismo , Miocárdio/patologia , Pericárdio/efeitos dos fármacos , Pericárdio/patologia , Ratos , Peixe-Zebra/crescimento & desenvolvimento
20.
Methods Mol Biol ; 2179: 79-106, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-32939715

RESUMO

The study of cell migration has been greatly enhanced by the development of new model systems and analysis protocols to study this process in vivo. Zebrafish embryos have been a principal protagonist because they are easily accessible, genetically tractable, and optically transparent. Neural crest cells, on the other hand, are the ideal system to study cell migration. These cells migrate extensively, using different modalities of movement and sharing many traits with metastatic cancer cells. In this chapter, we present new tools and protocols that allow the study of NC development and migration in vivo.


Assuntos
Movimento Celular/genética , Biologia Molecular/métodos , Crista Neural/ultraestrutura , Proteínas de Peixe-Zebra/ultraestrutura , Animais , Desenvolvimento Embrionário/genética , Peixe-Zebra/genética , Peixe-Zebra/crescimento & desenvolvimento
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