Alzheimer’s disease (AD) is a multifactorial neurodegenerative disorder with a high incidence rate in older people worldwide.^[1] Apart from its widely recognized hallmarks, such as β-amyloid (Aβ) plaques and hyperphosphorylated tau proteins, neuroinflammation, oxidative stress, and metal ion accumulation were found to severely contribute to the disease's progression.^[2] Within this intricate framework, sirtuin-1 (SIRT1) has gained attention as a potential AD preventive factor, since it is involved in oxidative stress responses and brain homeostasis.^[3] AD's high degree of complexity seems to justify the limited success of current single-target therapies and the recent shift of the research programmes towards multifunctional agents able to target multiple mechanisms simultaneously, with additive or even synergistic effects to more effectively counteract disease progression.

In this study, we present a series of previously reported SIRT1 activators featuring a 2,6-diphenylimidazo[1,2-a]pyridine core^[4] for potential anti-AD applications. We expanded the compound library by decorating the phenyl rings with various methoxy and/or hydroxy groups. Due to their polyphenolic nature, they showed antioxidant properties in vitro, as assessed in vitro through DPPH and ABTS assays and TBARS using rat brain homogenate. Also, although with specific preferences, some compounds showed metal-chelating abilities towards Cu²⁺, Zn²⁺, and Fe²⁺.

Notably, in differentiated SH-SY5Y cells, a recognized AD model, compound CLM400 was demonstrated to reduce oxidative stress when tested at 2.5 nM, whereas a pro-oxidant effect was observed at higher concentrations. Interestingly, TEM analyses revealed the compound's ability to form larger, non-toxic off-pathway Aβ aggregates with a pro-aggregating behavior. This finding is particularly relevant and aligns with previous studies on resveratrol, while traditional anti-AD agents often focus on inhibiting aggregation into toxic soluble oligomers.

This preliminary study highlighted a new series of multifunctional agents as promising agents for AD treatment. However, in-depth studies are required to shed light on the formed aggregates.

In recent years, there has been a significant increase in the chestnut processing industry, leading to the generation of substantial amounts of by-products that are causing disposal challenges and environmental consequences within the post-harvest supply chain of Castanea spp. nuts. In particular, chestnut episperm, produced in large quantities during post-harvest processing, shows potential to serve as a valuable source for extracting antioxidant compounds, including phenolics. This study has developed an environmentally sustainable method for extracting the polyphenols and antioxidants with best health-promoting properties from chestnut episperm (cv Marsol, C. sativa × C. crenata) using ultrasound technology. High-performance liquid chromatography (HPLC) was used to assess the levels of specific phenolic compounds known for their health benefits and antioxidant capacity. The total polyphenolic content (TPC) ranged from 90 to 150 mg GAE/g of dried weight (DW). Furthermore, a Ferric Reducing Antioxidant Power (FRAP) assay confirmed the high TPC levels in the extract, showing that all the extracts presented high levels of antioxidant properties (approximately 470–515 mmol Fe²⁺/Kg of dried weight for the most effective extracts). The proposed method enabled the production of preparations containing significant amounts of castalagin (20–80 mg/100 g DW), chlorogenic acid (15–25 mg/100 g DW), vescalagin (40–75 mg/100 g), and ferulic acid (80–120 mg/100 g DW). This investigation demonstrated the potential of chestnut episperm as an innovative source of antioxidants, which may be extracted using sustainable technologies and utilized in industry applications as pharmaceutical and/or food products, thereby promoting the valorization of agri-food waste through a low-impact reuse strategy and reducing environmental impacts. Chestnut companies stand to benefit financially by extracting antioxidant molecules through processing post-harvest wastes like episperm and marketing these preparations, thereby avoiding expenses associated with by-product disposal and potentially increasing their incomes.

Aging is a process characterized by a general decline in physiological functions. The high bioavailability of reactive oxygen species (ROS) plays an important role in the aging rate. Red blood cell deformability is the ability of cells to modulate their shape to ensure transit through narrow capillaries of the microcirculation. A loss of deformability can occur in several pathological conditions, during natural aging through an increase in membrane protein phosphorylation, and/or through the structural rearrangements of cytoskeletal proteins due to oxidative conditions, with a key role played by band 3. Due to the close relationship between aging and oxidative stress (OS), functional foods rich in flavonoids are excellent candidates to counteract age-related changes. This study aimed to verify the protective role of Açaì extract in a D-Galactose (D-Gal)-induced model of aging in human red blood cells. Our results showed that Açaì berry extract (10 μg/mL) avoided the formation of leptocytes, prevented the increased OS, and restored alterations in the distribution of band 3 protein (B3p) and CD47 proteins in red blood cells exposed to 100 mM D-Gal. Moreover, the significant decrease in membrane red blood cell deformability associated with D-Gal treatment was alleviated by pre-treatment with Açaì extract. The extract completely restored the increase in B3p phosphorylation and Syk kinase levels, but this effect was only partial for the alterations in the distribution of spectrin, ankyrin, and protein 4.1 after exposure to 100 mM D-Gal treatment. Interestingly, D-Gal exposure was also associated with an acceleration of the rate constant of SO42- uptake through B3p, as well as A1c formation. Both alterations were attenuated by pre-treatment with the Açaì extract. These findings contribute to clarifying the aging mechanisms in human red blood cells and to proposing flavonoid-rich functional foods as natural antioxidants for the treatment and prevention of OS-related disease conditions.

Rooibos (Aspalathus linearis) is a legume originally from Africa. Rooibos can come “unfermented” (green rooibos) or “fermented” (rooibos). The latter is obtained through an oxidation process controlling temperature and time. Apart from their different organoleptic properties, “fermented” and “unfermented” products also present different chromatographic profiles, with the dihydrochalcone aspalathin being the major antioxidant affected after processing.

In the present study, a commercial green rooibos powdered extract standardized to dihydrochalcones (aspalathin, nothofagin) and flavones (orientin, isorientin, vitexin…), together with the corresponding volatile fingerprint (safranal, damascenone…), was evaluated in a zebrafish model. The experimental groups were “control” (fed with commercial standard feed) and “Treatment” (fed with the standard feed including the green rooibos extract) groups. The protocol was performed twice, at 2 and 4 months post the start of ingestion, as follows: Each animal was individually placed at the bottom of a quadrangular glass aquarium (20x8x18 cm; length x width x depth; swimming volume: 3.5 L). The fish were filmed (1920 x 1080 px) for 6 min. The 1st minute was considered the time required to acclimate to the new environment, and the behavior analysis was considered for the remaining 5 min. Individual swimming activity was analyzed using Noldus Ethovision® tracking software, generating a virtual grid dividing the tank into two areas (upper and lower).

A standard diet including 0.05% green rooibos extract may modulate the behavior of zebrafish after 4 months of its continuous ingestion. This change in swimming patterns is associated with a state of lower anxiety in the zebrafish model. These results are supported by the trend towards lower latency to the first entry at the top of the tank.

Olive leaf extracts are rich in bioactive polyphenols, including secoiridoids, phenolic acids, and flavonoids, recognized for their anticancer potential. Among various cancers, breast cancer stands out as one of the most prevalent and lethal malignancies worldwide, with high morbidity and mortality rates. Despite advancements in treatment, including surgery, chemotherapy, hormone therapy, and targeted therapies, resistance to these interventions remains a significant challenge. As a result, there is a growing interest in plant-based compounds due to their potential to enhance the current therapies or provide novel, less toxic alternatives.

This study focuses on the antiproliferative effects of three flavonoids from olive leaves due to their well-documented antioxidant, anti-inflammatory, and anticancer properties. We aimed to determine how their combinations, at various ratios, influence the inhibition of breast cancer cell growth.

Using CompuSyn software, we modeled their synergistic, additive, and antagonistic effects based on the Combination Index values. Various ratios of the three flavonoids were tested through MTT viability assays on the MCF-7 (estrogen-receptor-positive) and JIMT-1 (HER2-positive, trastuzumab-resistant) breast cancer cell lines. Additionally, an 3D isobologram was generated to visualize and confirm the nature of these interactions. The results revealed that the type of interaction depends significantly on the flavonoid ratios.

The natural ratio in olive leaf extract demonstrated strong synergy, enhancing its antiproliferative activity and confirming prior research on its efficacy against breast cancer. The optimized flavonoid ratios showed even greater synergy, suggesting that tailored combinations could surpass the previously reported results and offer a new avenue for more potent, plant-based cancer therapies.

These findings underscore the importance of refining the flavonoid ratios and exploring their molecular mechanisms to enhance polyphenol-based treatments. Further research could lead to novel therapeutic formulations that provide more effective and targeted alternatives to the conventional treatments, improving patient outcomes while reducing side effects.

Introduction: Systemic Lupus Erythematosus (SLE) is a chronic autoimmune disease characterised by dysregulation of the immune system, production of self-reactive antibodies and an inflammatory state in organs and tissues. Oxidative stress has been implicated in the pathogenesis of cardiovascular events in SLE, promoting tissue injury and inflammation. Thrombosis is a well-known clinical feature in SLE but, despite the large number of risk factors, its underlying pathogenetic mechanisms are far to be understood. This study aimed to explore how oxidative changes in the structure and function of fibrinogen may contribute to the development of atherothrombosis in SLE.

Methods: Here, we recruited 144 adult SLE patients and 90 non-SLE controls. We assayed systemic redox status and, in purified fibrinogen fractions, we explored fibrinogen functional and structural features.

Results: Compared to the control group, SLE patients showed higher leukocyte ROS production. This increase was accompanied by higher plasma lipid peroxidation, decreased antioxidant defenses, and increased fibrinogen oxidation. Additionally, SLE patients displayed significant fibrinogen structural alterations and changes in blood clot architecture, characterized by lower porosity and a denser fibrin network with thinner fibrin fibers. Notably, functional properties of fibrinogen, such as thrombin-induced fibrin polymerization and susceptibility to plasmin-induced lysis, were more markedly affected in SLE patients than in controls. These structural and functional fibrinogen changes strongly correlated with redox parameters, which were also associated with SLE disease activity.

Conclusions: These data suggest that, in SLE patients, ROS induce structural and functional changes in fibrinogen. This could represent a new pathogenetic mechanism underlying atherothrombosis in SLE.

Introduction: Behçet’s syndrome (BS) is a systemic vasculitis of unknown etiology characterized as a multisystemic immune-inflammatory disorder affecting vessels of all sizes and frequently complicated by thrombosis. Systemic redox imbalance and heightened neutrophil activation in BS patients are believed to contribute to impaired coagulation. Our group have already demonstrated an altered fibrinogen structure and reduced fibrin susceptibility to plasmin-induced lysis in BS patients, primarily due to increased reactive oxygen species (ROS) production by neutrophils. This supports the idea that BS serves as a model for inflammation-driven thrombosis. Moreover, BS has been associated with a peculiar dysbiosis in the gut microbial ecosystem, represented by a depletion in the Clostridium cluster, involved in short-chain fatty acid (SCFA) production. Among SCFAs, patients show a decrease in the production of butyrate, which exerts anti-inflammatory and trophic activity.

Methods: To explore the relationship between butyrate, blood redox balance, and cardiovascular risk, we examined the effects of a 3-month butyrate-enriched diet in 30 bariatric surgery (BS) patients. Participants were randomly divided into two groups: 15 received oral butyrate supplementation (2.4 g/day), while 15 followed a lacto-ovo-vegetarian diet rich in insulin and resistant starch, which enhance butyrate production through fermentation. At baseline (T0) and after the intervention (T3), we assessed leukocyte intracellular ROS production, plasma malondialdehyde (MDA) levels as a marker of lipid peroxidation, and plasma total antioxidant capacity (TAC). Additionally, we evaluated fibrin susceptibility to plasmin-induced lysis using purified fibrinogen fractions.

Results: Both butyrate-enriched interventions led to significant reductions in leukocyte ROS production and plasma lipid peroxidation, alongside an increase in plasma total antioxidant capacity. Importantly, fibrin susceptibility to plasmin-induced lysis showed marked improvement in both groups at T3 compared to T0.

Conclusions: Our findings suggest that a butyrate-enriched diet enhances blood redox balance and promotes fibrin degradation, highlighting its potential role in cardiovascular disease prevention.

Introduction: Metabolic stress from prolonged hyperglycemia leads to adipocyte dysfunction, contributing to the pathogenesis of metabolic disorders such as obesity and type 2 diabetes. Numerous studies have shown that natural compounds are promising alternative therapies for alleviating metabolic alterations. However, their effects on human models are limited. Our objectives are to elucidate the metabolic alterations in human adipocytes under hyperglycemic conditions compared to mature adipocytes and to study the potential effect of compound A on antioxidant-related metabolism in hyperglycemic human adipocytes.

Methods: Hypertrophic human Simpson–Golabi–Behmel syndrome (SGBS) adipocytes, obtained by 2-week incubation with 25 mM of glucose, were used as a hyperglycemic model. Compound A was incubated during the last 48 h to study its effect on ROS production, on the expression of key target proteins like AKT (by Western blot), and on the modulation of metabolic pathways by profiling using UPLC-MS.

Results: Our findings revealed profound metabolic disruptions in adipocytes exposed to hyperglycemia, characterized by increased ROS, decreased AKT phosphorylation and adiponectin levels, and the downregulation of multiple metabolites and their associated pathways, like cysteine metabolism. Importantly, treatment with compound A demonstrated marked efficacy in mitigating these metabolic alterations and restoring adipocyte homeostasis. Compound A increased AKT phosphorylation and decreased ROS. Regarding metabolic pathways, it increased metabolites from cysteine and glutathione metabolism, partially reverting metabolic stress through an antioxidant environment.

Conclusions: These results provide insights into the metabolic pathways underlying adipocyte dysfunction in hyperglycemic conditions. In addition, they highlight the potential therapeutic effect of natural compound A in ameliorating metabolic disturbances associated with long-term hyperglycemia.

Introduction: Myelofibrosis (MF) is a myeloproliferative neoplasm characterized by clonal myeloproliferation, bone marrow fibrosis, extramedullary hematopoiesis, and abnormal inflammation. About 90% of MF patients carry mutations in the JAK2, CALR, or MPL genes. Among these, JAK2 mutations promote cytokine independence, constitutive activation of STAT proteins, and increased reactive oxygen species (ROS) production within hematopoietic stem cells. Excessive ROS and inflammation are potential triggers for thrombotic events, a leading cause of morbidity and mortality in myeloproliferative neoplasm patients.

To explore the mechanisms of inflammation-induced thrombosis in MF, we analyzed oxidation-induced fibrinogen alterations in MF patients compared to healthy controls and the effects of treatment with Ruxolitinib, a first-in-class JAK inhibitor.

Methods: This study included 20 primary MF patients and 20 age- and sex-matched healthy controls. Plasma redox status and structural and functional fibrinogen alterations were assessed in both primary MF patients (before and after Ruxolitinib treatment) and controls.

Results: MF patients displayed a significant increase in plasma lipid peroxidation and nitrate/nitrite levels, associated with a reduced total antioxidant capacity and free thiol plasma content. Oxidative imbalance was associated with fibrinogen oxidation, resulting in structural and functional alterations. Structural changes impaired fibrinogen polymerization into fibrin and reduced fibrin susceptibility to plasmin-induced lysis. A positive correlation between fibrin resistance to plasmin digestion and plasma oxidative stress was observed. Patients treated with Ruxolitinib exhibited significant improvements in redox status, as well as in fibrinogen structure and function.

Conclusions: MF patients exhibit a prothrombotic profile sustained by oxidative modifications of fibrinogen. Preliminary findings suggest that Ruxolitinib may improve redox balance and provide cardiovascular protection in these patients.

Obesity, a widespread global health challenge, is a condition characterized by an excessive accumulation of body fat that poses significant risks to overall health. It is often associated with a wide range of metabolic disorders, including insulin resistance, type 2 diabetes, dyslipidemia, hypertension, and cardiovascular diseases. Functional foods, particularly those enriched with bioactive compounds such as polyphenols and proteins, hold promise as effective tools for addressing obesity-related issues. Moreover, functional foods can address obesity-related comorbidities, such as cardiovascular disease and type 2 diabetes, by reducing markers like LDL cholesterol and fasting blood glucose levels [1,2]. This study evaluated the anti-obesity effects of hempseed-enriched kombucha (HK) using Caenorhabditis elegans as a model. The analysis included the quantification of reactive oxygen species (ROS) and the assessment of lipid storage using established staining techniques. First, HK was prepared with a previously developed formula [3] optimized to include hempseeds, and it was fermented for 12 days under controlled conditions until a safe pH (3.5-4.5) was achieved. Antioxidant capacity was analyzed via a 2,2-Diphenyl-1-picrylhydrazyl radical scavenging assay (DPPH+), Ferric Reducing Antioxidant Power (FRAP) assays, and a C. elegan in vivo assay, while fat storage reduction was assessed using Nile-Red fluorescence microscopy in daf-2 mutants. HK demonstrated significantly higher antioxidant activity (DPPH+ inhibition: 56%) compared to from conventional kombucha (K) (34% inhibition). HK significantly reduced reactive oxygen species (ROS) in C. elegans, showing lower fluorescence intensity in ROS assays compared to K. HK also reduced fat storage in C. elegans by 80-85%, outperforming K, which achieved a 60-65% reduction compared to glucose controls (p < 0.05). These findings suggest that the bioactive components in HK may enhance lipid metabolism through antioxidant mechanisms [4,5]. This research highlights the potential of hempseed-enriched kombucha as a functional beverage for managing obesity while contributing to sustainable health solutions.