INTRODUCTION: Pinenes, compounds from the monoterpene class, stand out for their bioactive properties, such as their antioxidant, anti-inflammatory, and antimicrobial effects. Oxidative stress, associated with various chronic diseases, has increased the demand for natural antioxidants like pinenes, which are widely used in the pharmaceutical and cosmetic industries. OBJECTIVE: We wished to investigate the antioxidant potential of pinenes. METHOD: This was a literature review on the antioxidant potential of pinenes. The article selection was conducted between October and November 2024 through searches in the Scielo, PubMed, and CAPES journal databases using the descriptors and the Boolean operators “pinene” AND “antioxidant.” RESULTS AND DISCUSSION: Research has shown that pinenes, such as alpha-pinene and beta-pinene, have significant antioxidant effects. Studies demonstrate that these compounds increase the activity of antioxidant enzymes, such as superoxide dismutase (SOD) and catalase (CAT), and reduce markers of oxidative stress, such as malondialdehyde (MDA) and reactive oxygen species (ROS). Alpha-pinene protects against carbon-tetrachloride-induced cardiac lesions in rats by modulating oxidative stress and inflammation. Beta-pinene has shown potential in reducing the oxidative stress caused by arsenic in rice plants, promoting the recovery of natural antioxidant defenses. Pinenes, especially alpha- and beta-pinene, demonstrate great potential as antioxidants, with the ability to reduce oxidative stress and modulate inflammatory processes. CONCLUSION: These results suggest that pinenes have therapeutic potential in diseases related to oxidative stress, such as neurodegenerative and inflammatory diseases, standing out as promising candidates for the development of treatments and products in the pharmaceutical and cosmetic industries. However, most of these studies have been conducted in animal or plant models, highlighting the need for human research to confirm these benefits.

Oxidative stress is an imbalance between the reactive oxygen species (ROS) and the endogenous antioxidant defense system in the body. Oxidative stress can injure cells in several ways, contributing to various diseases, such as atherosclerosis, cardiovascular disease, Alzheimer’s disease, and cancer. Lipoxygenase (LOX) is a family of non-heme iron-containing enzymes known for their catalysis of lipid peroxidation via the oxidation of polyunsaturated fatty acids (PUFAa). Oxidative stress can lead to the peroxidation of lipids, which can alter the substrate specificity and activity of LOX enzymes. This study aims to design benzimidazole derivatives and evaluate their antioxidant activity using an in silico approach. An oral bioavailability prediction was carried out using an online web server (ADMETlab 2.0). The compounds were designed, optimized, and prepared using ChemDraw 12.0, Spartan14, and UCFS Chimera. The molecular docking of the compounds in the LOX enzyme was carried out using AutoDock Vina. All the designed ligands passed Lipinski’s rule of five, which means that they are all druggable. All the ligands have a good synthetic accessibility score (< 6). The docking results showed that the ligands have a good binding affinity to the target, the LOX enzyme (1N8Q), with L20 having the best binding affinity (-6.0 Kcal/mol). The native ligand (DHB: 3,4-DIHYDROXYBENZOIC ACID) has a binding score of -5.7 Kcal/mol. Ascorbic acid was used as a reference drug and had a binding score of -5.4 Kcal/mol. The results indicated that ligands interacted with the binding pockets of the enzyme’s active site through conventional hydrogen bond, hydrogen bond, Van der Waals, Pi-Pi, Pi-alkyl, pi-anion, and alkyl interactions. The interacting amino acid residues responsible for the scavenging activity are HIS518, GLN514, LEU515, HIS523, ALA561, TRP519, and ILE572. This study suggested that designed benzimidazole ligands have the potential to scavenge free radicals.

The impact of 6-hydroxy-L-nicotine on inflammation in a 5xFAD mouse model of Alzheimer’s disease

Alexandra Mara Cimpanu¹, Lucian Hritcu¹, Razvan Stefan Boiangiu¹

¹BioActive Research Group, Department of Biology, Alexandru Ioan Cuza University of Iasi, Romania

Introduction: Alzheimer’s disease (AD) is a neurodegenerative disorder and the most common form of dementia. AD patients suffer from memory loss, aphasia, and personality and behavior changes and exhibit difficulties in thinking, language, and problem-solving skills. The main neuropathologic features of AD include extracellular plaques containing amyloid beta (Aβ) and intracellular neurofibrillary tangles of hyperphosphorylated tau protein. In addition to these essential features, several other pathologic changes such as inflammation and the sustained activation of microglia and other immune cells are commonly associated with AD. Nicotine has been reported to reduce anxiety and improve memory, learning, and attention, but its therapeutic use in AD is limited by its side effects. Here, we aim to evaluate the anti-inflammatory effect of a structure-related nicotine derivative, namely 6-hydroxy-L-nicotine (6HLN), in a transgenic mouse model of AD.

Methods: 6HLN was chronically administered in doses of 0.3 and 0.6 mg/kg (b.w., i.p., for 30 days) to a 5xFAD mouse model of AD. By using ELISA, we measured the levels of glial fibrillary acidic protein (GFAP), nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB), interleukin 6 (IL-6), and tumor necrosis factor α (TNF-α) in the brains of the 6HLN-treated mice.

Results: Our results showed that 6HLN treatment reduced, in a dose-dependent manner, the levels of GFAP, NF-kB, IL-6, and TNF-α in the brain of the transgenic mouse model of AD, indicating a promising anti-inflammatory potential of 6HLN.

Conclusion: These findings are in accordance with those from the literature, indicating that 6HLN might represent a new neuropharmacological agent in ameliorating AD. This work was supported by CNCSIS-UEFISCSU, project number PN-III-P4-PCE-2021-1692.

Redox homeostasis in the human body is strictly regulated by reducing molecules, such as glutathione, as well as various antioxidant enzymes. A correlation between oxidative stress damage and the majority of diseases has already been shown. Therefore, studying the antioxidant effects of natural products with prominent antioxidative properties is necessary for the prevention and treatment of various pathological conditions. RJ is a bee product known for its abundance in bioactive substances and its function as a very potent regulator of many metabolic processes. It is also considered a medicinal agent that can cope with oxidative stress. Being easy to access and produce, and due to its great antioxidant properties, RJ has become an ideal option for helping oncological patients that undergo chemotherapy, which can induce severe levels of oxidative stress in body.

This study aimed to evaluate RJ’s ability to scavenge reactive species and modulate the GSTP1 marker in healthy cells in order to offer a scientific basis for the further development and use of this natural bee product.

Healthy lung fibroblasts (MRC-5 cell line) were treated with RJ at a concentration of 100 μg/mL, and after 24 h, the level of O₂˙ˉ superoxide anion radicals was assessed by an NBT test, while the gene expression of the GSTP1 marker was analyzed by the quantitative real-time polymerase chain reaction (qPCR) method.

This natural bee product reduced the concentration of O₂˙ˉ in the tested cells within 24 h of treatment, and this antioxidative effect was most likely due to upregulated GSTP1 expression. The protein product of the GSTP1 gene is responsible for the catalyzation of glutathione (GSH) binding to oxidative stress metabolites, such as superoxide anion radicals, and their further neutralization in cells.

We found that RJ has an important protective effect against the oxidative damage of human lung fibroblast cells, and its properties can be used to explore new resources for pharmacological treatments, as well as to improve natural medicine.

During cell aging, mitochondrial ROS (mtROS) cause mitochondrial DNA mutations, which lead to mitochondrial dysfunction. This causes an increased production of mtROS, creating a vicious cycle and leading to accelerated aging. We used rotenone, a complex I inhibitor, to cause mitochondrial dysfunction in dermal fibroblasts, which resulted in high mitochondrial and cytosolic ROS levels, apoptotic cell death, increased matrix metalloproteinase-1 secretion, and decreased collagen secretion. Pre-treatment with carotenoid-rich tomato extracts, rosemary extract, and estradiol reversed these effects. The aim of the current study was to evaluate the effect of rotenone and phytonutrients and estradiol on mitochondrial respiration and cell senescence and to analyze the mechanisms involved in these effects. Rotenone caused a substantial reduction in mitochondrial respiration and ATP levels and increased the number of senescent cells. These effects were reversed by phytonutrients and estradiol. The enhanced mitochondrial respiration as a result of the phytonutrient and estradiol treatment was associated with increased mitochondrial biogenesis. Rotenone treatment was associated with an increased activity of NFƙB and AP-1 transcription systems, whereas the phytonutrients and estradiol caused an inhibition of these activities and an activation of antioxidant response element (ARE/Nrf2) transcriptional activity. To determine whether the activation of ARE/Nrf2 and inhibition of NFƙB are crucial for cell protection, we inhibited these pathways by the Nrf2 inhibitors ML385 and ochratoxin A and by the NFƙB inhibitors IKK-16 and JSH-23. Inhibition of ARE/Nrf2 markedly reduced the protective effects of the phytonutrients and estradiol by diminishing their potential to reduce mtROS and the ATP level. Inhibition of NFƙB markedly reduced rotenone-induced cell damage, similar to the effects of the phytonutrients and estradiol, suggesting that NFkB inhibition is important for skin cell protection. The presented results indicate that tomato carotenoids, rosemary extract, and estradiol protect dermal fibroblasts from damage caused by mtROS and may thus delay skin cell senescence and improve skin health.

Cistus ladanifer L. is a Mediterranean shrub and is widely distributed in the Iberian Peninsula. It has high importance in the perfume industry and is used as a medicinal plant due to its essential oils. Its residues from distilleries, however, are underutilized. These residues are potential sources of phenolic compounds that can be extracted and used in several applications. In this work, extracts from distillery residues were produced using two methods, solid–liquid extraction (SLE) and subcritical water extraction (SWE). Two extracts were produced for each method, under different conditions. For the SLE extraction, the two treatment options were CLL40°C (1g:50 mL 50:50 water:methanol (v/v), 1 h extraction time, 40 °C) and CLL60°C (1g:100 mL 50:50 water:methanol (v/v), 1 h extraction time, 60 °C). For the SWE extraction, the extracts were produced by mixing 2 g of CLL with 200 mL of ultrapure water at 60 bar and 100 or 150 °C. The yield of the extraction was assessed, along with its total phenolic content (TPC) and antioxidant activity being assessed using different assays (DPPH^• and ABTS^•+ scavenging activities and FRAP assay). CLL40°C displayed the highest TPC (175.24 ± 21.82 mg gallic acid equivalents/g dry extract) and the strongest antioxidant activity in the DPPH^• and ABTS^•+ assays (267.81 ± 60.11 and 518.90 ± 50.16 mg Trolox equivalents/g dry extract). CLL60°C displayed the highest results in the FRAP assay (11.64 ± 2.96 mg ascorbic acid equivalents/g dry extract). The analysis of the SWE extracts is currently being performed and will be finished in time for the conference. These results will offer valuable information on the impact and potential of different extraction methods for the valorization of labdanum residues and phenolic extraction.

Rising awareness about the advantages of natural antioxidants has driven efforts to explore alternative natural sources for these compounds. Macroalgae are a promising source of these bioactive compounds. Brown macroalgae are renowned for their ecological significance, metabolite diversity, and potent bioactive attributes. These metabolites, spanning polysaccharides, phenolic compounds, and terpenoids, play a dual role by influencing the ecological dynamics of the algae and exhibiting noteworthy bioactive characteristics, highlighting their antioxidant potential as the common driver of their effects. This study aimed to characterize four brown macroalgae by comparing two orders, Fucales (Fucus vesiculosus and Pelvetia canaliculata) and Laminariales (Laminaria ochroleuca and Saccharina latissima), searching for bioactive compounds, namely, pigments and phlorotannins, with antioxidant properties, and to later optimize their extraction by Heat-Assisted Extraction (HAE) and Pressurized Liquid Extraction (PLE). The experiments were designed following a response surface methodology (RSM). Extraction was performed according to the following independent variables: for HAE, time (t, 5-60 min), temperature (T, 30-90°C) and solvent concentration (S, 0-100% ethanol) and for PLE, time (t, 1-50 min), temperature (T, 50-200°C) and ethanol percentage of the solvent (S, 0-100%). Phenolic compounds and pigments were identified by HPLC-ESI-QqQ-MS/MS following the previous methodology. In both cases, PLE improved the extraction of active compounds. The extraction yield of F. vesiculosus – PLE ranged from 9.1 to 63.4%, obtaining the best ratio among the tested species. Phlorotannins and carotenoids were the most frequent compounds to exhibit antioxidant properties. The presence of quinic acid and scopoletin was also characterized. This work will shed light on the optimization and potential of these algae-derived compounds, paving the way for further research in natural antioxidants for industrial application.

Optimization of antioxidant extraction through artificial neural networks

The increasing demand in modern society for natural sources of bioactive compounds with potential applications in preventive medicine has driven the development of innovative approaches to optimize extraction processes. While Response Surface Methodology (RSM) has been extensively utilized for prediction and optimization in extraction studies, Artificial Neural Networks (ANNs) provide a powerful alternative by enabling the development of nonlinear computational models. These models are capable of self-learning and training to address practical challenges, without relying on predefined mathematical equations, by mimicking the structure and functionality of biological neural networks. The application of ANNs offers numerous advantages, including the ability to interpret complex datasets, scale results through optimization and parallelization, and model complex, nonlinear relationships. Furthermore, ANNs allow for the handling of large datasets and generalization across systems, without the limitations of predefined models or specific experimental designs. This systematic review compiles studies on the application of ANNs in the optimization of extraction processes of antioxidants present in natural matrices (e.g., edible plants, vegetables, and fruits), providing an objective evaluation of their potential for the sustainable development of industrial products enriched with these compounds. Special attention is given to how ANNs outperform traditional techniques, such as RSM, in predicting yields, enhancing extraction efficiency, and minimizing resource consumption. By exploring key studies and methodologies, this review aims to highlight the role of ANNs in advancing green and sustainable technologies, offering novel insights into their applicability in designing industrial processes that incorporate natural antioxidants into food, cosmetics, and pharmaceutical products.

This patent landscape study provides a detailed overview of innovation in antioxidants derived from Lamiaceae plants, offering valuable insights into current trends and future opportunities for research and industrial development. As medicinal preparations, antioxidants from plants such as rosemary, thyme, and lavender are increasingly used in treatments targeting oxidative stress and related health conditions. The primary focus lies on the antioxidant properties of Lamiaceae plants, which have gained significant attention for their ability to scavenge free radicals, providing vital protection against oxidative damage.

To identify patents related to plant-based antioxidants derived from the Lamiaceae family, we followed an approach based on the intersection of different Cooperative Patent (CPC) codes, which enables the identification of relevant patents by combining therapeutic applications, medicinal preparations, and botanical classifications. The results of this analysis were then presented in terms of publication trends, jurisdictional distribution, key players, classification overview, and technological focus within market dynamics.

The data indicate a steady increase in the number of patents published from 2001 to 2021. The analysis shows a diverse distribution of key players across regions. China leads in terms of both the number of applicants and innovation diversity, followed by the United States and Europe. Asia’s robust representation underscores the global importance of Lamiaceae-derived antioxidants in various industrial sectors. The presence of European players indicates a steady focus on applications with high commercial value, such as cosmetics and functional foods, while North American companies drive advancements in pharmaceuticals and nutraceuticals. The CPC subgroup analysis demonstrates that patents for antioxidants from Lamiaceae plants are mainly concentrated in medicinal, dermatological, and cosmetic applications. The versatility of these plants supports innovation in pharmaceuticals, skincare, and wellness products, with a growing trend towards natural and sustainable solutions in various industries.

Antioxidants from seafood play a crucial role in promoting human health by providing a natural source of compounds that mitigate oxidative stress. Oxidative stress, usually as a result of antioxidant imbalance, is strongly linked with DNA damage, inflammation, metabolic diseases, and other hazardous events. This intricate process can be ameliorated by endogenous antioxidant enzymes and molecules, but also by intake of antioxidants naturally present in food. Seafood is rich in bioactive antioxidants like astaxanthin, tocopherols, selenium, coenzyme Q10, glutathione, and omega-3 fatty acids, all of which have demonstrated significant potential in reducing cellular oxidative damage.

Some antioxidants in fish, such as astaxanthin, tocopherols, and selenium, collaborate in mitigating oxidative stress by neutralizing free radicals. This action helps to reduce cellular damage, thereby lowering the risk of chronic diseases such as cancer, cardiovascular diseases, and neurodegenerative disorders. Astaxanthin, a carotenoid found in certain fish species like salmon and trout, is known for its superior antioxidant capacity, surpassing vitamin E and other carotenoids. Tocopherols (vitamin E compounds) also contribute to the prevention of lipid oxidation in cell membranes, enhancing cellular integrity and function. Selenium, an essential trace mineral in fish, is a component of selenoproteins with critical antioxidant functions, protecting against oxidative damage and supporting immune function. Additional antioxidants such as coenzyme Q10, found in fish like sardines and mackerel, contribute to cellular energy production while neutralizing free radicals. Glutathione, present in various types of seafood, supports detoxification pathways and bolsters cellular antioxidant defense systems.

This review synthesizes the mechanisms, health benefits, and dietary implications of seafood-derived antioxidants, providing insights into their potential for enhancing human health.