IntroductionGlioblastoma (GB) is the most aggressive primary glioma, with a median survival of 15-18 months. Current treatments are often ineffective, largely due to tumor heterogeneity and recurrence. Advances in understanding GB’s molecular landscape and microenvironment have highlighted new therapeutic strategies to fight this life-threatening tumor. Given the pivotal role of natural compounds in drug discovery, those with anti-inflammatory and cytotoxic/cytostatic properties are emerging as promising candidates for GB therapy.MethodsThis study investigates the antitumor and immunomodulatory effects of 7α-acetoxy-6β-hydroxyroyleanone (Roy), a diterpene isolated by our team from Plectranthus hadiensis Schweinf., using both 2D and 3D GB cell models. U87 cells were used as a standard GB model and to generate monocellular and multicellular spheroids (U87, HMC3, and/or HBMEC cells). Both models were treated with 16 µM of Roy, a concentration previously shown to be tumor-specific.ResultsRoy significantly reduced spheroid size and metabolic activity over time, with the most pronounced effects observed in multicellular spheroids. This compound also inhibited cell proliferation by preventing colony formation and downregulating CDK4 and VEGFA mRNA levels. Roy’s bioactivity was enhanced in the presence of conditioned medium (secretome from GB and/or microglia cells), exerting a neuromodulatory effect by modulating IL6/JAK2/STAT3 mRNA expression and by suppressing the secretion of cytokines involved in the chronic inflammatory state within the GB microenvironment. Importantly, Roy was also able to cross the blood-brain barrier.ConclusionThese findings, in line with our previous work, underscore the cytotoxic potential of this natural compound, suggesting Roy as a promising lead candidate for future GB treatment strategies.

Plant-based extracts are rich sources of phenolic compounds, which may act as skin antiaging mediators. Herein, Cistus albidus L. (Ca), Cistus ladanifer L. subsp. ladanifer (Cl) and Cistus salviifolius L. (Cs) were selected to test whether their phytochemical profile and bioactive potential align to target human skin aging. Hydroethanolic extracts (HEs) were prepared and characterized using infrared vibrational spectroscopy (FTIR-ATR) and liquid chromatography-mass spectrometry (LC-MS). Non-toxic concentrations were screened, and cytoprotective and antioxidant effects were studied in tert-butyl hydroperoxide-stimulated normal human dermal fibroblasts (NHDFs). Lipopolysaccharide-stimulated RAW 264.7 macrophages were used to assess anti-inflammatory activity, the Organization for Economic Co-operation and Development (OECD) Test Guideline No. 439 was used to assess irritant effects, and the anti-senescence potential was assessed in etoposide-stimulated NHDFs. A series of enzymatic inhibition assays was performed. All extracts comprised ellagic acid derivatives, as well as myricetin and quercetin derivatives in Cs and Ca. The HE of Cs was also markedly composed of ligstroside. At non-toxic concentrations, cytoprotective effects were observed in NHDFs. However, only Cs and Cl exhibited significant antioxidant activity in these cells (p < 0.001 and p < 0.0001, respectively). In addition to that, Cl demonstrated highly significant anti-inflammatory (p < 0.0001) and anti-senescence (p < 0.0001) effects. Cs and Cl showed a remarkable potential to inhibit elastase; in addition, Cs also showed anti-hyaluronidase and anti-tyrosinase activities. Meaningfully, Cs and Cl extracts did not exhibit skin irritant effects. The unveiled potential of Cl in skin aging offset highlights the need to elucidate the detailed mechanisms of action, paving the way for the development of skin anti-aging formulations.

Understanding the neural mechanisms underlying the impact of psychedelics on social perception and cognition may be instrumental to unravel their therapeutic potential. We conducted a pharmacoimaging study to examine ayahuasca's effects on a key theory of mind region, at the core of the third visual pathway (TVP)-the posterior superior temporal sulcus (pSTS), which is involved in facial emotion recognition and social perception. Twelve healthy participants (mean age: 40 ± 6.6 years; four females) completed a crossover design with three conditions: 0.5 mg/kg N, N-dimethyltryptamine (DMT), 0.8 mg/kg DMT, and placebo, with 1-2 months washout intervals. Resting-state functional magnetic resonance imaging (fMRI) was used to assess pSTS functional and effective connectivity. The highest dose significantly increased right pSTS connectivity and directed modulation from visual (primary and extrastriate cortices) and mirror neuron regions (supplementary motor cortex; SMC). Subjectively, this enhanced social cognitive states, with a strong positive correlation between pSTS-SMC connectivity and perspective-taking experiences. Additionally, ayahuasca produced positive psychological effects, including improved perceived social relationships, at 1-week follow-up despite minimal acute effects. Our findings reveal a novel mechanism of action of psychedelics at early stages of social information processing, with enhanced integration of the TVP and mirror neuron systems. The pSTS emerged as a critical hub supported by top-down and bottom-up evidence, providing a basis for understanding ayahuasca's prosocial therapeutic effects.

Metabolic dysfunction-associated steatotic liver disease (MASLD) is an increasing public health burden closely linked to oxidative stress and mitochondrial dysfunction. We evaluated the protective effects of Equisetum ramosissimum extracts, ethanolic (ER_EtOH80) and decoction (ER_D), against palmitic acid (PA)-induced lipotoxicity in HepG2 cells. PA treatment markedly increased intracellular lipid droplets (LDs), raising droplet number to 331 % and total LD area to 498 % of control (CTL). Both extracts prevented LD enlargement, reducing mean LD size to 165 % of PA values. ER_D improved fatty acid oxidation, elevating CPT1 alpha to 376 % of CTL and HADHA to 145 % of PA, and restored PA-impaired respiration, recovering basal, maximal, and ATP-linked oxygen consumption. ER_D also enhanced mitochondrial biogenesis, increasing mitochondrial DNA copy number (136 % of CTL) and upregulating proteins such as VDAC (126 % of CTL). In parallel, ER_EtOH80 mitigated oxidative stress, lowering ROS by 17 % and raising glutathione by 20 % vs PA, while inducing Nrf2 targets NQO1 and HMOX1. Antioxidant enzymes were differentially regulated: ER_EtOH80 increased catalase activity to 125 % of CTL, whereas ER_D elevated SOD2 levels (147 % of CTL). Both extracts preserved AMPK alpha signaling, stimulated autophagy (LC3II/LC3I ratio, similar to 200 % of CTL), and prevented PA-induced lysosomal loss (similar to 40 % recovery vs PA). In summary, Equisetum ramosissimum extracts protect hepatocytes through complementary mechanisms, with ER_D enhancing mitochondrial function and biogenesis and ER_EtOH80 reinforcing antioxidant defenses, ultimately restoring cellular quality control under lipotoxic stress and highlighting their potential in MASLD prevention.

Background/Objectives: Plants inhabiting mediterranean-influenced climatic zones, like Helichrysum stoechas (L.) Moench subsp. stoechas, Lavandula pedunculata (Mill.) Cav., and Thymus mastichina (L.) L. subsp. mastichina, have been scarcely investigated regarding their richness in phenolic compounds, herein explored as sources of skin anti-aging compounds. Methods: In this investigation, Fourier transform infrared spectroscopy (FTIR) in attenuated total reflectance (ATR) mode and high-performance liquid chromatography coupled with diode-array detection and electrospray ionization tandem mass spectrometry (HPLC-DAD-ESI/MSn) were employed to chemically characterize the hydroethanolic extracts (HEs), and their cell-free antioxidant potential was screened. Thereafter, non-toxic concentrations of HEs were determined in human skin cells using Alamar blue® and Sulforhodamine B assays. The cytoprotective and antioxidant effects of HEs were assessed in tert-butyl hydroperoxide-stimulated fibroblasts, their anti-inflammatory potential was studied in lipopolysaccharide-injured macrophages, and enzymatic inhibition assays were performed. Notably, the irritant effects of HEs were tested according to Test Guideline No. 439 of the Organization for Economic Co-operation and Development (OECD). Results: The major compounds identified in the T. mastichina and L. pedunculata HEs were rosmarinic and salvianolic acid derivatives, while H. stoechas HE was mainly composed of caffeoyl and feruloyl derivatives, and O-glycosylated flavonoids. T. mastichina (≤0.4 mg/mL) exhibited significant cytoprotective, anti-inflammatory, and antioxidant effects, as well as remarkable anti-hyaluronidase activity. Conclusions: Shedding light on the quantitative and qualitative chemical picture of these HEs highlighted T. mastichina as a promising candidate to target skin aging effects, which correlates with its phenolic content. Further investigation is warranted regarding its anti-aging pharmacological activity, which could lead to the development of plant-based skin anti-aging products.

This study aimed to investigate β-Caryophyllene (BCA) pharmacokinetics as well as the potential antitumor activity and mechanism of action of BCA and eugenol (EU), alone or in combination, in U87 glioblastoma (GB) cells. The BCA pharmacokinetic was studied by evaluating its concentration profiles in rat blood and cerebrospinal fluid after oral and intravenous administration. EU and BCA antitumor mechanisms were assessed by comparing their effects in U87 GB cells and non-tumoral HMC3 cells. Cell death, cell cycle regulation and mitochondrial membrane potential (MMP) were evaluated using flow cytometry. mRNA levels of target genes were evaluated by qPCR. Secreted cytokines were measured by Luminex®. BCA, as well as EU, permeates the brain. EU and BCA affected the viability and proliferation of U87 cells (up to 50%, p < 0.001) but not HMC3 cells and showed a synergistic effect. BCA and EU induced G0/G1 cell cycle arrest, increasing apoptosis/necrosis. EU and BCA induced the downregulation of mRNAs encoding for key proteins involved in GB angiogenesis (VEGFA decreased op to 60%, p < 0.01), proliferation and progression, and showed anti-inflammatory activity (IL-4 significantly decreased, p < 0.001). EU and BCA demonstrated strong and multitarget antitumor activity in U87 cells. Our results provide a strong rationale for the further evaluation of EU and BCA as possible therapeutic molecules in GB management.

Background/Objectives: Fungal keratitis (FK) is a current challenge in ophthalmology due to its association with severe visual impairment and the limitations of current antifungal therapies. We aim to evaluate the antifungal activity of essential oils (EOs) from the aromatic and medicinal plants Cymbopogon citratus and Lavandula pedunculata against selected FK pathogens collected from FK patients in two Portuguese hospitals. Methods: The antifungal activity of the EOs was tested at concentrations of 25%, 50%, 75%, and 100% for up to 7 days using the solid-phase disk diffusion in vitro assay. Results: Candida albicans was the most prevalent pathogen (28.6%), followed by Candida parapsilosis (21.4%) and Dicyma olivacea (14.2%). The other identified species were Aspergillus fumigatus and Scedosporium boydii (7.1%). Clinical diagnostic methodologies showed agreement with the molecular identification. Cymbopogon citratus EO showed higher antifungal activity than Lavandula pedunculata EO. The highest antifungal activity was observed against Aspergillus fumigatus and Scedosporium boydii (inhibition zone diameter, IZD = 90.0 mm) after 7 (Cymbopogon citratus EO) or 3 days of incubation (Lavandula pedunculata EO). While the antifungal activity of Cymbopogon citratus EO was maintained during the study (for Aspergillus fumigatus, Candida albicans, and Scedosporium boydii), the antifungal activity of Lavandula pedunculata EO decreased with time. Conclusions: Cymbopogon citratus EO and Lavandula pedunculata EO showed optimal antifungal activity against molds (Aspergillus fumigatus and Scedosporium boydii) after 3 days of incubation. Against yeasts (Candida albicans and Candida parapsilosis), the EOs showed lower activity. Our study sheds light on the development of new pharmacological strategies for FK based on EOs extracted from aromatic and medicinal plants.

Ketoprofen (KET) has been proven effective against glioblastoma. Nevertheless, its hydrophobic nature and susceptibility to degradation upon administration, added to the blood-brain barrier's low permeability, make its repurposing for brain cancer treatment challenging. Having this in mind, the purpose of this work was to develop KET-loaded in situ thermoreversible nanoemulgels for intranasal nose-to-brain delivery, aiming at increased drug strength and protection, controlled drug release, and improved bioavailability. Formulations containing Capryol (R) 90, Tween (R) 80, Transcutol (R) HP, poloxamer 407, and water were produced through spontaneous emulsification. Formulations' droplet size, polydispersity index (PDI), zeta potential, pH, rheology, stability, in vitro drug release, and in vitro antitumor efficacy and safety were evaluated. A high drug strength (4 mg/mL), and small (20-30 nm) and monodisperse (PDI 0.1-0.2) nanodroplets, with slightly negative to neutral zeta potential (-1.5 to-10 mV), were obtained. The nanoemulgels' also revealed skin adequate pH (approximate to 6), nasal cavity temperature' sol-gel transitions (32 degrees C), elevated viscosity (13660-927302 cP), and high cumulative controlled in vitro drug release (approximate to 78-93 %), following Makoid-Banakar and Weibull kinetic models. Optimized nanoemulgels revealed relevant efficacy against human glioblastoma U87 cells. Therefore, intranasal thermoreversible KETloaded nanoemulgels were successfully developed, of innovative composition, showing promising results for glioblastoma treatment. Given their high scalability potential, the developed nanoplatforms could be promising candidates for translational applications, and future in vivo assays could further confirm their potential, so that they might one day be considered as an adjuvant or primary treatment for glioblastoma.

Reward processing is a broad psychological construct that can be parsed into distinct components known as “reinforcement learning” (learning), “reward responsiveness” (liking), and “motivation to obtain a reward” (wanting). Dysfunctions in reward processing in mesolimbic and mesocortical pathways are a core feature of many pathologies. Psychedelics have been proposed as a treatment option for multiple disorders affecting the reward system, but mechanistic studies are lacking. In this preliminary, hypothesis-generating pharmacoimaging study, we evaluated the effects of inhaled N, N-dimethyltryptamine (DMT) with a particular focus on the connectivity of the mesocorticolimbic circuitry. Our within-subject pharmacoimaging design included 11 healthy participants with prior experience in psychedelics. In the active condition, DMT was self-administered immediately before MRI acquisition, while in the control condition there was no administration. We found decreased connectivity between the right nucleus accumbens (NAc) and the left ventral tegmental area (VTA), increased connectivity between the right NAc and anterior cingulate cortex (ACC) and increased connectivity between the medial prefrontal cortex (mPFC) and the ACC. These results correlated with changes in volition and perception, as measured with the hallucination rating scale. In sum, we found reduced connectivity in the midbrain-NAc pathway, which connectivity is often increased in addiction, and increased connectivity between reward/affective regions and the ACC. These findings suggest a potential therapeutic potential of psychedelics in disorders affecting reward processing.

As is the case for all forms of cultural heritage, museum collections are under a serious threat of biodeterioration, with fungi playing a significant role in this phenomenon, compromising the preservation and dissemination of past knowledge, artifacts and traditions. As part of the ongoing efforts to study the fungal contaminations observed in the artifacts that compose the collection of the Science Museum of the University of Coimbra, non-invasive sampling of four objects present in the archive was conducted and 43 fungal isolates were retrieved. These isolates were subjected to a comprehensive analysis based on molecular and morphological characteristics and were identified as belonging to 15 different species. Of the 15 isolated species, 5 were selected for in vitro antifungal essential oil assays, based on their relative abundance and biodeteriorative potential. For these, essential oils of Cymbopogon citratus, Lavandula angustifolia, Thapsia gummifera, Mentha pulegium, Mentha suaveolens, Thapsia villosa, Thymus mastichina and Agastache foeniculum were obtained via hydrodistillation, in accordance with the European Pharmacopeia. Of the tested oils, most impacted fungal growth, with Cymbopogon citratus and Lavandula angustifolia being able to fully inhibit the growth of most fungi even at the lowest tested concentrations.

Background/Objectives: Côa Valley, located in the northeast of Portugal, harbors more than 500 medicinal plant species. Among them, four species stand out due to their traditional uses: Equisetum ramosissimum Desf. (hemorrhages, urethritis, hepatitis), Rumex scutatus L. subsp. induratus (Boiss. and Reut.) Malag. (inflammation, constipation), Geranium purpureum Vill., and Geranium lucidum L. (pain relief, gastric issues). Given their rich ethnomedicinal history, we evaluated their protective effects on an in vitro model of metabolic dysfunction-associated steatotic liver disease (MASLD). Methods: Decoction (D) and hydroalcoholic (EtOH80%) extracts were prepared and chemically characterized. Their safety profile and effects on lipid accumulation were assessed in palmitic acid (PA)-treated HepG2 cells using resazurin, sulforhodamine B, and Nile Red assays. Results: Chemical analysis revealed diverse phenolic compounds, particularly kaempferol derivatives in E. ramosissimum. All extracts showed minimal cytotoxicity at 25–50 µg/mL. At 100 µg/mL, only E. ramosissimum extracts maintained high cell viability. In the lipotoxicity model, E. ramosissimum decoction demonstrated the most potent effect, significantly reducing PA-induced neutral lipid accumulation in a dose-dependent manner, while other extracts showed varying degrees of activity. Conclusions: These findings highlight E. ramosissimum’s decoction, rich in kaempferol derivatives, as particularly effective in reducing lipid accumulation in this MASLD cell model while also providing a comprehensive characterization of traditionally used plants from the Côa Valley region.