Multilevel assessment of carbamazepine effects: An integrative approach using zebrafish early-life stages.

Carbamazepine (CBZ) is the most commonly used drug in epilepsy treatment, and its metabolites are commonly detected among persistent pharmaceuticals in the aquatic environment. This study aimed to investigate CBZ effects on early-life-stage zebrafish (Danio rerio) (from 2 to 168 hpf) by employing of an integrative approach linking endpoints from molecular to individual level: (i) development; (ii) locomotor activity; (iii) biochemical markers (lactate dehydrogenase, glutathione-S-transferase, acetylcholinesterase and catalase) and (iv) transcriptome analysis using microarray. A 168 h - LC50 of 73.4 mg L-1 and a 72 h - EC50 of 66.8 mg L-1 for hatching were calculated while developmental effects (oedemas and tail deformities) were observed at CBZ concentrations above 37.3 mg L-1. At the biochemical level, AChE activity proved to be the most sensitive parameter, as evidenced by its decrease across all concentrations tested (∼25% maximum reduction, LOEC (lowest observed effect concentration) < 0.6 µg L-1). Locomotor behaviour seemed to be depressed by CBZ although this effect was only evident at the highest concentration tested (50 mg L-1). Molecular analysis revealed a dose-dependent effect of CBZ on gene expression. Although only 25 genes were deregulated in organisms exposed to CBZ when compared to controls, both 0.6 and 2812 µg L-1 treatments impaired gene expression related to development (e.g. crygmxl1, org, klf2a, otos, stx16 and tob2) and the nervous system (e.g. Rtn3, Gdf10, Rtn3), while activated genes were associated with behavioural response (e.g. prlbr and taar). Altogether, our results indicate that environmentally relevant CBZ concentrations might affect biochemical and genetic traits of fish. Thus, the environmental risk of CBZ cannot be neglected, especially in a realistic scenario of constant input of domestic effluents into aquatic systems.

Hurdles in translating science from lab to market in delivery systems for Cosmetics: An industrial perspective.

In recent decades, a sweeping technological wave has reshaped the global economic landscape. Fueled by the unceasing forces of digital innovation and venture capital investment, this transformative machine has left a significant mark across numerous economic sectors. More recently, the emergence of 'deep tech' start-ups, focusing on areas such as artificial intelligence, nanotechnology, and biotechnology, has infused a fresh wave of innovation into various sectors, including the pharmaceutical and cosmetic industry. This review explores the significance of innovation within the cosmetics sector, with a particular emphasis on delivery systems. It assesses the crucial process of bridging the gap between research and the market, particularly in the translation of nanotechnology into tangible real-world applications. With the rise of nanotechnology-based beauty ingredients, we can anticipate groundbreaking advancements that promise to surpass consumer expectations, ushering in a new era of unparalleled innovation in beauty products.

Oily core/amphiphilic polymer shell nanocapsules change the intracellular fate of doxorubicin in breast cancer cells.

The aim of this work was to develop and test the in vitro biological activity of nanocapsules loaded with a doxorubicin (DOX) free base dissolved in a core of castor oil shelled by poly(methyl vinyl ether-co-maleic anhydride) conjugated to n-octadecylamine residues. This system was stable and monodisperse, with a hydrodynamic diameter of about 300 nm. These nanocapsules changed the intracellular distribution of DOX, from the nuclei to the cytoplasm, and exhibited higher toxicity towards cancer cells - 4T1 and MCF-7 - and significantly lower toxicity towards normal cells - NIH-3T3 and MCF-10A - in vitro. In conclusion, these nanocapsules are suitable DOX carriers, which remain to be studied in in vivo tumor models.

CNTs coated charcoal as a hybrid composite material: Adsorption of fluoxetine probed by zebrafish embryos and its potential for environmental remediation.

Although traditional water treatment systems can remove various substances from wastewater, these conventional systems fail to remove many chemical molecules that pose potential ecological and health risks. Carbon nanotubes (CNTs) appear attractive to adsorption of many substances, but CNTs adsorbed with toxic substances becomes a nanocomposite still more toxic. Here, we employ zebrafish embryos as biosensor to examine how a hybrid micro/nanostructured carbonaceous material (HMNC) derived from a combination of activated carbon (AC) with hydrophilic carbon nanotubes (CNTs) can remediate wastewater contaminated with the pharmaceutical fluoxetine hydrochloride (FLX). AC and HMNC are practically non-toxic to zebrafish embryos (LC50 > 1000 mg.L-1). HMNC addition to culture medium containing FLX significantly reduces sublethal effects and lethality. Interaction between FLX and HMNC involves chemical adsorption such that embryo co-exposure to HMNC adsorbed with FLX in the range of concentrations evaluated herein does not elicit any behavioral changes in zebrafish.

Exposure to dilute concentrations of bupropion affects zebrafish early life stages.

Psychiatric pharmaceuticals are one of the most prescribed active substances globally. Bupropion (BPP) is an antidepressant that acts via inhibition of norepinephrine and dopamine reuptake. It has been found in various water matrices, and thus its effects on aquatic organisms must be studied. The present study aimed to evaluate the acute toxic effects of BPP on zebrafish (Danio rerio) early life stages. For developmental analysis, organisms were exposed for 168 h to concentrations ranging from 0 to 82000 µg/L. Two other experiments were performed by exposing embryos to a wide range of concentrations (from 0 to 50000 µg/L) in order to evaluate BPP effects on embryonic behavior, using the Zebrabox and testing at the biochemical level (acetylcholinesterase, glutathione-S-transferase, lactate dehydrogenase and catalase). Developmental analysis indicated that BPP had low acute toxicity with a calculated 168 h-LC50 of 50346 µg/L. Concentrations equal to or above 44800 µg/L elicited several effects such as hatching delay, edemas and tail deformities. However, concentrations from 7300 µg/L upwards elicited equilibrium alteration. Behavioral analysis showed that BPP affected zebrafish locomotor behavior by decreasing activity at 0.6 µg/L, increasing activity at 8.8 and 158 µg/L, and decreasing activity at 50000 µg/L. Biochemical analysis showed an increase of AChE activity at 158 and 2812 µg/L, an increase in GST at the highest concentrations, CAT alteration and increase of LDH at 0.6, 2812 and 50000 µg/L. We can conclude that BPP affects zebrafish early life stages at environmental concentrations.

Exposure to low concentration of fluoxetine affects development, behaviour and acetylcholinesterase activity of zebrafish embryos.

Fluoxetine (FLX) is a selective serotonin reuptake inhibitor (SSRI) antidepressant widely used in clinics and very often found in environmental samples of urban aquatic ecosystems in concentrations ranging from ng/L to µg/L. Fish populations might be especially susceptible to FLX due to the presence of conserved cellular receptors of serotonin. Neurotoxic effects on fish biota of polluted water bodies may be expected, but there are no sufficient studies in the current literature to elucidate this hypothesis. Batteries of embryo larval assays with zebrafish were performed to evaluate the potential effects of FLX exposure, including environmentally relevant concentrations. Evaluated parameters included survival, development, behaviour and neuronal biochemical markers. Regarding acute toxicity, a 168 h-LC50 value of 1.18 mg/L was obtained. Moreover, hatching delay and loss of equilibrium were observed, but at a concentration level much higher than FLX measured environmental concentrations (>100 µg/L). On the other hand, effects on locomotor and acetylcholinesterase activity (≥0.88 and 6 µg/L, respectively) were found at levels close to the maximum reported FLX concentration in surface waters. Altogether, these results suggest that FLX is neurotoxic to early life stages of zebrafish, in a short period of time causing changes in important ecological attributes which can probably be linked from molecular to population level.

Chronic effects of carbamazepine on zebrafish: Behavioral, reproductive and biochemical endpoints.

Carbamazepine (Cbz), one of the most prescribed pharmaceuticals in the world is often detected in surface waters and sediments. However, few studies addressed its chronic effects in fish. In the present study, Danio rerio adults were exposed for 63 days to Cbz (0 - control, 10 µg L-1 - concentration found in effluents, and 10,000 µg L-1 - 5% of LC50 at 72 h). Assessed endpoints were: feeding behavior, growth rate, number of eggs produced and their viability, histological alterations in female gonads, and biochemical biomarkers associated with antioxidant defenses (catalase - CAT, and glutathione S-transferase - GST activities), neurotransmission (acetylcholinesterase activity - AChE) and metabolism (lactate dehydrogenase - LDH). Cbz exposure increased the total time for food intake but did not affect D. rerio growth. Although the total number of eggs was not affected by Cbz exposure, the eggs viability was significantly impaired. Exposure to Cbz caused alterations in the female gonads follicular stages. In terms of biochemical endpoints, CAT activity in liver and gills, was sensitive to the pharmaceutical exposure presenting a decreased activity. AChE activity was induced in the head (both concentrations) and muscle (10,000 µg L-1). GST activity was increased in gills (both concentrations) but inhibited in the intestine. Concerning LDH, enzymatic activity was increased in the liver and decreased in muscle and gills. Several of the above-mentioned effects can be directly linked with effects at population level (e.g. feeding behavior) and occurred at environmental concentrations (the lowest concentration tested), thus serious concerns regarding risks posed by Cbz residues to fish populations arise with this study.

Selol nanocapsules with a poly(methyl vinyl ether-co-maleic anhydride) shell conjugated to doxorubicin for combinatorial chemotherapy against murine breast adenocarcinoma in vivo.

Nanocapsules containing selol and doxorubicin (NCS-DOX) with an oily core of selol and a shell of poly(methyl vinyl ether-co-maleic anhydride) covalently conjugated to doxorubicin were developed in a previous work. In this study, these nanocapsules showed a similar antitumour effect in comparison to the free doxorubicin (DOX) treatment, but showed no evident DOX-related cardiotoxicity, as evidenced by serum creatine kinase-MB (CK-MB) activity. The histopathological analysis showed that the free DOX treatment induced more intense morphological damage to myocardial tissues in comparison to NCS-DOX treatment. Animals treated with free DOX presented important muscle fibre degradation and animals treated with NCS-DOX, heart tissue did not present signals of muscle fibre degeneration. These results indicate that the cardiotoxicity related to DOX is reduced when this drug is carried by the NCS-DOX. Noteworthy, biodistribution analyses showed that NCS-DOX accumulated more intensely in tumours than the free DOX. Thus, this study reinforces the importance of the development of nanocapsules as drug carriers for the treatment of cancer.