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- Development of dual-crosslinked Pluronic F127/Chitosan injectable hydrogels incorporating graphene nanosystems for breast cancer photothermal therapy and antibacterial applicationsPublication . Pouso, Manuel António do Rosário; Melo, Bruna L.; Gonçalves, Joaquim; Mendonça, António; Correia, I.J.; Diogo, Duarte de MeloNanomaterials with responsiveness to near-infrared light can mediate the photoablation of cancer cells with an exceptional spatio-temporal resolution. However, the therapeutic outcome of this modality is limited by the nanostructures’ poor tumor uptake. To address this bottleneck, it is appealing to develop injectable in situ forming hydrogels due to their capacity to perform a tumor-confined delivery of the nanomaterials with minimal off-target leakage. In particular, injectable in situ forming hydrogels based on Pluronic F127 have been emerging due to their FDA-approval status, biocompatibility, and thermosensitive sol–gel transition. Nevertheless, the application of Pluronic F127 hydrogels has been limited due to their fast dissociation in aqueous media. Such limitation may be addressed by combining the thermoresponsive sol–gel transition of Pluronic F127 with other polymers with crosslinking capabilities. In this work, a novel dual-crosslinked injectable in situ forming hydrogel based on Pluronic F127 (thermosensitive gelation) and Chitosan (ionotropic gelation in the presence of NaHCO3), loaded with Dopamine-reduced graphene oxide (DOPA-rGO; photothermal nanoagent), was developed for application in breast cancer photothermal therapy. The dual-crosslinked hydrogel incorporating DOPA-rGO showed a good injectability (through 21 G needles), in situ gelation capacity and cytocompatibility (viability > 73 %). As importantly, the dual-crosslinking improved the hydrogel’s porosity and prevented its premature degradation. After irradiation with near-infrared light, the dual-crosslinked hydrogel incorporating DOPA-rGO produced a photothermal heating (ΔT ≈ 22 °C) that reduced the breast cancer cells’ viability to just 32 %. In addition, this formulation also demonstrated a good antibacterial activity by reducing the viability of S. aureus and E. coli to 24 and 33 %, respectively. Overall, the dual-crosslinked hydrogel incorporating DOPA-rGO is a promising macroscale technology for breast cancer photothermal therapy and antimicrobial applications.
- The role of ayahuasca in cell viability and oxidative stress in gastric adenocarcinoma cell linePublication . Gonçalves, Joana; Cascalheira, José; Valentão, Patrícia; Luís, Ângelo; Gallardo, Eugenia; Duarte, Ana PaulaAyahuasca, a psychoactive beverage native to the Amazon, originally derived from Banisteriopsis caapi stem scrapings and Psychotria viridis leaves, exhibits hallucinogenic properties due to N,Ndimethyltryptamine. When combined with β-carbolines, it enters the bloodstream and central nervous system, inhibiting monoamine oxidase-A. Over time, therapeutic effects have been associated to ayahuasca consumption. This study assessed the impact of extracts from three plant decoctions used in ayahuasca preparation on the gastric adenocarcinoma cell line (AGS). MTT reduction assays selected B. caapi, Mimosa hostilis, and Peganum harmala samples as most effective. Lactate dehydrogenase activity evaluated membrane integrity loss, while oxidative stress induction was measured using dihydroethidium and 2′,7′-dichlorodihydrofluorescein diacetate probes. Results revealed apoptosis induction in AGS Ucells, with all three samples significantly reducing oxidative stress.
- Pullulan-Tween 40 emulsified films containing geraniol: production and characterization as potential food packaging materialsPublication . Simões, Alexandra; Ramos, Ana; Domingues, F.C.; Luís, ÂngeloPetroleum-based plastics have been widely used as packaging materials because of their low-cost availability and good mechanical properties. However, the use of plastics has become restricted as they are highly resistant to biodegradation, causing environmental problems. This work aimed to produce and characterize emulsified pullulan films incorporating geraniol for application as food packaging materials with potential to substitute the conventional plastics. When geraniol was incorporated in the films, they showed antimicrobial activity against Enterococcus faecalis ATCC 29212 (inhibition zone diameter = 15.19 ± 0.66 mm) and Pseudomonas aeruginosa ATCC 27853 (inhibition zone diameter = 10.99 ± 1.82 mm). Furthermore, scanning electron microscopy showed the inhibition of Enterococcus faecalis ATCC 29212 biofilms when they were directly formed on the emulsified pullulan films incorporating geraniol. The produced films also demonstrated high transparency (> 90%) and hydrophilic surfaces (water contact angle < 90°). This work demonstrated the viability of using geraniol to produce pullulan active films as new food packaging materials.
- Action of Curcumin on Glioblastoma Growth: A Systematic Review with Meta-Analysis of Animal Model StudiesPublication . Luís, Ângelo; Amaral, Leonor; Domingues, F.C.; Pereira, L.; Cascalheira, JoséGliomas are aggressive brain tumors with poor prognosis even after surgical removal and radio-chemotherapy, stressing the urgency to find alternative therapies. Several preclinical studies evaluating the anticancer effect of curcumin in animal models of glioma are reported, but a systematic review with meta-analysis of these studies, considering the different experimental conditions used, has not been made up to this date. A search in different databases (Pubmed, Web of Science, Scopus, and SciELO) following the PRISMA statement was conducted during November 2023 to systematically identify articles assessing the effect of curcumin in murine xenograft models of glioma and identified 15 articles, which were subdivided into 24 studies. Tumor volume before and after treatment with curcumin or vehicle was extracted and the efficacy of curcumin was evaluated by performing a random effects meta-analysis of the data. Publication bias and the impact of different experimental conditions on curcumin efficacy were assessed. Treatment with curcumin decreased tumor volume. Comparing curcumin with control groups, the overall weighted standardized difference in means was −2.079 (95% CI: −2.816 to −1.341; p-value < 0.001). The curcumin effect was observed for different animal models, types of glioma cells, administration routes, and curcumin formulations. Publication bias was identified but does not invalidate curcumin’s effectiveness. The findings suggest the potential therapeutic efficacy of curcumin against glioma.
- Hyaluronic acid-functionalized graphene-based nanohybrids for targeted breast cancer chemo-photothermal therapyPublication . Lima-Sousa, Rita; Melo, Bruna L.; Mendonça, António; Correia, I.J.; Melo-Diogo, Duarte deNanomaterials’ application in cancer therapy has been driven by their ability to encapsulate chemotherapeutic drugs as well as to reach the tumor site. Nevertheless, nanomedicines’ translation has been limited due to their lack of specificity towards cancer cells. Although the nanomaterials’ surface can be coated with targeting ligands, such has been mostly achieved through non-covalent functionalization strategies that are prone to premature detachment. Notwithstanding, cancer cells often establish resistance mechanisms that impair the effect of the loaded drugs. This bottleneck may be addressed by using near-infrared (NIR)-light responsive nanomaterials. The NIR-light triggered hyperthermic effect generated by these nanomaterials can cause irreversible damage to cancer cells or sensitize them to chemotherapeutics’ action. Herein, a novel covalently functionalized targeted NIR-absorbing nanomaterial for cancer chemo-photothermal therapy was developed. For such, dopamine-reduced graphene oxide nanomaterials were covalently bonded with hyaluronic acid, and then loaded with doxorubicin (DOX/HA-DOPA-rGO). The produced nanomaterials showed suitable physicochemical properties, high encapsulation efficiency, and photothermal capacity. The in vitro studies revealed that the nanomaterials are cytocompatible and that display an improved uptake by the CD44-overexpressing breast cancer cells. Importantly, the combination of DOX/HA-DOPA-rGO with NIR light reduced breast cancer cells’ viability to just 23 %, showcasing their potential chemo-photothermal therapy.
- Pretreated Agro-Industrial Effluents as a Source of Nutrients for Tomatoes Grown in a Dual Function Hydroponic System: Tomato Quality AssessmentPublication . Afonso, Alexandra; Ribeiro, Carlos; Carvalho, Maria João; Correia, Tânia; Correia, Pedro; Regato, Mariana; Costa, Idália; Fernandes, Annabel; Almeida, Adelaide; Lopes, Ana; Carvalho, FátimaIn a zero-waste approach for the agro-industrial sector, this study aimed to evaluate the reuse of cheese whey wastewater (chemical oxygen demand = 2.1 g L−1) pretreated by immediate one-step lime precipitation followed by natural carbonation as a nutritive solution for tomato production in hydroponic systems. Pretreated effluent, diluted with groundwater (1:6) and supplemented with nutrients, was utilized to irrigate different hydroponic systems designed to assess the influence of tomato rooting type (free/confined−setup_A) and the feed’s solution level (with/without water deep−setup_B). Plants and fruit development, fruit physicochemical characteristics and sensory analysis, and effluent quality after reuse were analyzed. Good quality tomato production with high crop yield was obtained. The highest marketable tomato weight per plant (682 g) was observed in setup_B with a deep-bed system, but setup_A, with free or confined rooting, presented similar values. The type of rooting, within setup_A or water deep within setup_B, did not significantly influence plant and fruit characteristics. The highest maturity and flavor indexes were observed for setup_A with free rooting. Regarding sensory analysis, setup_A often scored the highest in terms of overall appreciation with free or confined rooting. The reuse of cheese whey wastewater in hydroponics reduced freshwater consumption for crop production, allowed for a treated final effluent and prevented soil degradation in a sustainable circular economy methodology.
- Reuse of Pretreated Agro-Industrial Wastewaters for Hydroponic Production of LettucePublication . Afonso, Alexandra; Regato, Mariana; Patanita, Mariana; Luz, Silvana; Carvalho, Maria João; Fernandes, Annabel; Lopes, Ana; Almeida, Adelaide; Costa, Idália; Carvalho, FátimaThe utilization of agro-industrial wastewaters (AIWWs), pretreated by immediate one-step lime precipitation + natural carbonation, as a nutritive solution for the hydroponic production of lettuce was evaluated. The AIWWs studied were olive mill wastewater (OMW), winery wastewater (WW), and cheese whey wastewater (CWW). Lettuces (Lactuca sativa L. var. crispa) were grown in a closed nutrient film technique hydroponic system, using the pretreated AIWWs (OMW-T, WW-T, and CWW-T) and a control nutrient solution (CNS). The growth and sensory analysis of lettuces and the environmental parameters of effluents after hydroponics were evaluated. The average number of lettuce leaves obtained with nutrient solutions prepared with AIWW-T was lower than that from CNS, but the highest lettuce chlorophyll content was attained with CWW-T, which also presented the best grow results. In general, sensory analysis did not show significant differences from the lettuces grown in the different pretreated AIWWs and CNS. As for the environmental parameters of the effluents from hydroponics, according to the Portuguese legislation, only the chemical oxygen demand of the OMW-T and WW-T presented slightly higher values than that of the environmental limit values for discharge in surface waters, showing the feasibility of using pretreated agro-industrial effluents in hydroponic lettuce cultivation, while obtaining a treated effluent, in a circular economy perspective.
- Reuse of Textile Dyeing Wastewater Treated by ElectrooxidationPublication . Pinto, Cláudia; Fernandes, Annabel; Lopes, Ana; Nunes, Maria João; Baía, Ana; Ciríaco, Lurdes; Pacheco, Maria JoséWastewater reuse has been addressed to promote the sustainable water utilization in textile industry. However, conventional technologies are unable to deliver treated wastewater with the quality required for reuse, mainly due to the presence of dyes and high salinity. In this work, the feasibility of electrooxidation, using a boron-doped diamond anode, to provide treated textile dyeing wastewater (TDW) with the quality required for reuse, and with complete recovery of salts, was evaluated. The influence of the applied current density on the quality of treated TDW and on the consecutive reuse in new dyeing baths was studied. The ecotoxicological evaluation of the process towards Daphnia magna was performed. After 10 h of electrooxidation at 60 and 100 mA cm−2, discolorized treated TDW, with chemical oxygen demand below 200 (moderate-quality) and 50 mg L−1 (high-quality), respectively, was obtained. Salt content was unchanged in both treatment conditions, enabling the consecutive reuse without any salt addition. For the two reuse cycles performed, both treated samples led to dyed fabrics in compliance with the most restrictive controls, showing that an effective consecutive reuse can be achieved with a moderate-quality water. Besides the water reuse and complete salts saving, electrooxidation accomplished an ecotoxicity reduction up to 18.6-fold, allowing TDW reuse without severe ecotoxicity accumulation.
- Electrochemical Recovery of Phosphorus from Simulated and Real Wastewater: Effect of Investigational Conditions on the Process EfficiencyPublication . Sousa, Carlos Y.; Fernandes, Annabel; Amaro, Albertina; Pacheco, Maria José; Ciríaco, Lurdes; Lopes, AnaThe development of recovery processes has become essential in recent years as a strategy to minimize environmental pollution while boosting circular economy and sustainable development. Due to the exponential growth in agricultural production and the increased pollution of waterbodies, the production of fertilizers from recovered phosphorus has become an alternative to phosphate rock-based production. In this work, the effect of different operational parameters in the efficiency of the electrochemical recovery of phosphorus, from organic and inorganic sources, was investigated. Among the studied variables, the most significant was the electrode material utilized in the system. The use of magnesium sacrificial electrodes, as AZ31 alloys, led to phosphorus removal from solution of above 90%, allowing the recovery of both orthophosphates and organic phosphorus (glyphosate) as struvite, brucite, and other amorphous compounds. Since there is a lack in the literature about the use of magnesium electrodes in real wastewater electrochemical treatment, system efficiency was also evaluated using a sanitary landfill leachate, reaching 96% of phosphorus recovery. The specific energy consumption and faradaic efficiency of the phosphorus recovery process were also assessed.
- Therapeutic Potential of Resveratrol for Glioma: A Systematic Review and Meta-Analysis of Animal Model StudiesPublication . Luís, Ângelo; Marcelino, Helena; Domingues, F.C.; Pereira, L.; Cascalheira, JoséGliomas are aggressive malignant brain tumors, with poor prognosis despite available therapies, raising the necessity for finding new compounds with therapeutic action. Numerous preclinical investigations evaluating resveratrol’s anti-tumor impact in animal models of glioma have been reported; however, the variety of experimental circumstances and results have prevented conclusive findings about resveratrol’s effectiveness. Several databases were searched during May 2023, ten publications were identified, satisfying the inclusion criteria, that assess the effects of resveratrol in murine glioma-bearing xenografts. To determine the efficacy of resveratrol, tumor volume and animal counts were retrieved, and the data were then subjected to a random effects meta-analysis. The influence of different experimental conditions and publication bias on resveratrol efficacy were evaluated. Comparing treated to untreated groups, resveratrol administration decreased the tumor volume. Overall, the effect’s weighted standardized difference in means was −2.046 (95%CI: −3.156 to −0.936; p-value < 0.001). The efficacy of the treatment was observed for animals inoculated with both human glioblastoma or rat glioma cells and for different modes of resveratrol administration. The combined administration of resveratrol and temozolomide was more effective than temozolomide alone. Reducing publication bias did not change the effectiveness of resveratrol treatment. The findings suggest that resveratrol slows the development of tumors in animal glioma models.