Toxicological and anti-tumor effects of a linden extract (Tilia platyphyllos Scop.) in a HPV16-transgenic mouse model

Tilia platyphyllos Scop. is a popular broad-leaved tree, native to Central and Southern Europe. Hydroethanolic extracts rich in phenolic compounds obtained from T. platyphyllos Scop. have shown in vitro antioxidant, anti-inflammatory and antitumor properties. The aim of this work was to evaluate the therapeutic properties of a hydroethanolic extract obtained from T. platyphyllos in HPV16-transgenic mice. The animals were divided into eight groups according to their sex and phenotype. Four groups of female: HPV+ exposed to linden (HPV linden; n = 6), HPV+ (HPV water; n = 4), HPV- exposed to linden (WT linden; n = 5) and HPV- (WT water; n = 4) and four groups of male: HPV+ exposed to linden (HPV linden; n = 5), HPV+ (HPV water; n = 5), HPV- exposed to linden (WT linden; n = 5) and HPV- (WT water; n = 7). The linden (Tilia platyphyllos Scop.) extract was orally administered at a dose of 4.5 mg/10 mL per animal (dissolved in water) and changed daily for 33 days. The hydroethanolic extract of T. platyphyllos consisted of protocatechuic acid and (-)-epicatechin as the most abundant phenolic acid and flavonoid, respectively, and was found to be stable during the studied period. In two male groups a significant positive weight gain was observed but without association with the linden extract. Histological, biochemical, and oxidative stress analyses for the evaluation of kidney and liver damage support the hypothesis that the linden extract is safe and well-tolerated under the present experimental conditions. Skin histopathology does not demonstrate the chemopreventive effect of the linden extract against HPV16-induced lesions. The linden extract has revealed a favourable toxicological profile; however, additional studies are required to determine the chemopreventive potential of the linden extract. This journal isThis work was supported by the project IBERPHENOL, project number 0377_IBERPHENOL_6_E; Interact R&D project, operation number NORTE-01-0145-FEDER-000017, National Funds by FCT – Portuguese Foundation for Science and Technology, under the project UIDB/04033/2020 (CITAB), and project UIDB/ CVT/00772/2020 (CECAV) and the post-graduation grant SFRH/ BD/136747/2018 and 2020.04789.BD; the authors are also grateful to FCT, Portugal and FEDER under programme PT2020 for financial support to CIMO (UIDB/00690/2020) and L. Barros acknowledges the national funding by FCT, P. I., through the institutional scientific employment program-contract. The authors would like to thank Cantinho das Aromáticas organic farmers from Vila Nova de Gaia (Portugal) for providing the samples. This work was financially supported by: Base Funding - UIDB/00511/2020 of the Laboratory for Process Engineering, Environment, Biotechnology and Energy – LEPABE - funded by national funds through the FCT/MCTES (PIDDAC).info:eu-repo/semantics/publishedVersio

Inflammatory, Oxidative Stress, and Apoptosis Effects in Zebrafish Larvae after Rapid Exposure to a Commercial Glyphosate Formulation

Glyphosate-based herbicides (GBH) are the most used herbicides in the world, carrying potentially adverse consequences to the environment and non-target species due to their massive and inadequate use. This study aimed to evaluate the effects of acute exposure to a commercial formulation of glyphosate, Roundup® Flex (RF), at environmentally relevant and higher concentrations in zebrafish larvae through the assessment of the inflammatory, oxidative stress and cell death response. Transgenic Tg(mpxGFP)i114 and wild-type (WT) zebrafish larvae (72 h post-fertilisation) were exposed to 1, 5, and 10 µg mL−1 of RF (based on the active ingredient concentration) for 4 h 30 min. A concentration of 2.5 µg mL−1 CuSO4 was used as a positive control. Copper sulphate exposure showed effectiveness in enhancing the inflammatory profile by increasing the number of neutrophils, nitric oxide (NO) levels, reactive oxygen species (ROS), and cell death. None of the RF concentrations tested showed changes in the number of neutrophils and NO. However, the concentration of 10 µg a.i. mL−1 was able to induce an increase in ROS levels and cell death. The activity of antioxidant enzymes (superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx)), the biotransformation activity, the levels of reduced (GSH) and oxidised (GSSG) glutathione, lipid peroxidation (LPO), lactate dehydrogenase (LDH), and acetylcholinesterase (AChE) were similar among groups. Overall, the evidence may suggest toxicological effects are dependent on the concentration of RF, although at concentrations that are not routinely detected in the environment. Additional studies are needed to better understand the underlying molecular mechanisms of this formulation

Ocorrência de Aeromonas sp. e microrganismos psicrotróficos e estimativa do prazo de validade comercial de filé de pintado (Pseudoplatystoma coruscans) mantidos sob refrigeração

Verificou-se a ocorrência de bactérias do gênero Aeromonas e estimou-se o prazo de validade comercial de filés de pintado (Pseudoplatystoma coruscans) com pele, durante estocagem em refrigeração por meio da quantificação de microrganismos heterotróficos aeróbios psicrotróficos e análises físico-químicas para determinação do pH e detecção de amônia e gás sulfídrico. Foram utilizadas 45 amostras de filé de pintado, com aproximadamente 100 gramas cada, embaladas individualmente em polietileno de alta densidade e armazenadas em câmara frigorífica entre 0ºC e 3ºC. A cada dois a três dias de estocagem, três unidades de filés foram submetidas a análises microbiológicas e físico-químicas, totalizando 15 análises durante o período de estocagem. As contagens de Aeromonas sp. e microrganismos heterotróficos aeróbios psicrotróficos variaram de 1,89 a 9,47logUFC/g e 0 a 6,54logUFC/g, respectivamente. A variação do pH foi de 6,20 a 6,97, e as análises de amônia e gás sulfídrico foram negativas durante todo o período. O pH dos filés de pintado atingiu o limite máximo de 6,4 aos 23 dias de estocagem, e estimou-se o seu prazo de validade comercial

Effects of Glyphosate or Glyphosate-Based Herbicide during the Zebrafish Life Cycle: A Review Addressing the Mechanisms of Toxicity

Herbicides with glyphosate (GLY) as an active ingredient (a.i.) are increasingly used, and GLY is currently the most used herbicide in the world. Consequently, its residues have often been found in aquatic ecosystems. Investigating how this substance affects aquatic species is a priority in ecotoxicology research, especially in fish, as they can absorb and concentrate toxins. In this sense, a critical review was performed, synthesizing data from the peer-reviewed bibliography, reporting on the toxicity of exposure to pure GLY and glyphosate-based herbicides (GBHs), using zebrafish as an animal model. The concentrations of this herbicide that induced toxic effects are highly variable, with some exceeding the limits determined by regulatory agencies. Globally, relevant toxic effects have been reported in zebrafish, namely, teratogenic effects incompatible with life, which translates directly into an increase in reported zebrafish mortality. Neurotoxicity, genotoxicity, changes in energy metabolism and oxidative stress, and immune and hormonal system dysfunction with an impact on fish reproduction were also described. In conclusion, both GLY and GBHs may induce damage to zebrafish, compromising their survival, reproduction, and maintenance. These results may be valid and applied to other fish species and aquatic ecosystems

Anti-Inflammatory, Anti-Oxidative and Anti-Apoptotic Effects of Thymol and 24-Epibrassinolide in Zebrafish Larvae

Thymol (THY) and 24-epibrassinolide (24-EPI) are two examples of plant-based products with promising therapeutic effects. In this study, we investigated the anti-inflammatory, antioxidant and anti-apoptotic effects of the THY and 24-EPI. We used zebrafish (Danio rerio) larvae transgenic line (Tg(mpxGFP)i114) to evaluate the recruitment of neutrophils as an inflammatory marker to the site of injury after tail fin amputation. In another experiment, wild-type AB larvae were exposed to a well known pro-inflammatory substance, copper (CuSO4), and then exposed for 4 h to THY, 24-EPI or diclofenac (DIC), a known anti-inflammatory drug. In this model, the antioxidant (levels of reactive oxygen species—ROS) and anti-apoptotic (cell death) effects were evaluated in vivo, as well as biochemical parameters such as the activity of antioxidant enzymes (superoxide dismutase, catalase and glutathione peroxidase), the biotransformation activity of glutathione-S-transferase, the levels of glutathione reduced and oxidated, lipid peroxidation, acetylcholinesterase activity, lactate dehydrogenase activity, and levels of nitric acid (NO). Both compounds decreased the recruitment of neutrophils in Tg(mpxGFP)i114, as well as showed in vivo antioxidant effects by reducing ROS production and anti-apoptotic effects in addition to a decrease in NO compared to CuSO4. The observed data substantiate the potential of the natural compounds THY and 24-EPI as anti-inflammatory and antioxidant agents in this species. These results support the need for further research to understand the molecular pathways involved, particularly their effect on NO