valorization-red-seaweed-gracilaria-gracilis-through-biorefinery

Valorization of the Red Seaweed <em>Gracilaria gracilis</em> Through a Biorefinery Approach

The interest in seaweeds as an abundant feedstock to obtain valuable and multitarget bioactive ingredients is continuously growing. In this work, we ...

Polytechnic of Leiria

Polytechnic University of Leiria

 is a marine brown seaweed mainly found on the Atlantic coast. Herein, we report the antioxidant and neuroprotective activities of seven fractions (F1⁻F7) obtained by normal phase chromatography from the  dichloromethane extract, as well as of its two major isolated diterpenes. Total phenolic content of fractions was determined by the Folin⁻Ciocalteu method, while antioxidant activity was evaluated by the DPPH, ORAC, and FRAP assays. Neuroprotective effects were evaluated in a neurotoxic model induced by 6-hydroxydopamine (6-OHDA) in a human neuroblastoma cell line (SH-SY5Y), while the mechanisms associated to neuroprotection were investigated by the determination of mitochondrial membrane potential, H₂O₂ production, Caspase-3 activity, and by observation of DNA fragmentation. Fractions F4 and F5 exhibited the best neuroprotective and antioxidant activities, respectively. F4 fraction prevented changes in mitochondrial potential, and induced a reduction of H₂O₂ levels production and an increase in cell viability, suggesting that it may contain multi-target compounds acting on different pathways. Hence, this fraction was subjected to purification steps, affording the known diterpenes eleganolone and eleganonal. Both compounds exhibited antioxidant potential, being interesting candidates for further neuroprotective studies.

Antioxidant and Neuroprotective Potential of the Brown Seaweed  in an in vitro Parkinson's Disease Model.

Due to the current demand for healthier food, the IV gamma products industry has grown over the years. Additionally, it is well known that chlorine is a disinfectant largely used in this industry. However, the risk assessment associated with the formation of carcinogenic by-products from this halogenated disinfection has not been widely studied. The aim of this study was to assess the trihalomethanes (THMs) exposure through the consumption of IV gamma products, more specifically, salad mixes, and potential health risk in Portuguese population using a deterministic method. The quantification of THMs in salad mixes from the most representative Portuguese brands was performed by gas chromatography coupled to mass spectrometry (GC-MS). The THMs exposures were estimated by combining the THMs concentration data with a ready-to-eat salad mix consumption survey (case study applied in the Lisboa e Vale do Tejo area; n = 271). The concentration of THMs in all samples was below the limit of quantification of the equipment (1 μg/L), and this limit was used for the evaluation of the exposure. It was found that it is very small for the consumption pattern of these products in the studied population categories, suggesting that there is no risk for consumers due to the halogenated disinfection process. The highest estimated daily exposure was 7,6510 mg/ kg bw/ week, which corresponds to a person who consumes such products daily.

Risk assessment of trihalomethanes exposure by consumption of IV gamma products: Evidences from a Portuguese regional survey.

Parkinson's disease (PD) is the second most common neurodegenerative disorder, and is characterized by a progressive degeneration of the dopaminergic neurons in the . Although not completely understood, several abnormal cellular events are known to be related with PD progression, such as oxidative stress, mitochondrial dysfunction and apoptosis. Accordingly, the aim of this study was to evaluate the neuroprotective effects of  enriched fractions in a neurotoxicity model mediated by 6-hydroxydopamine (6-OHDA) on SH-SY5Y human cells, and the disclosure of their mechanisms of action. Additionally, a preliminary chemical screening of the most promising bioactive fractions of  was carried out by GC-MS analysis. Among the tested fractions, four samples exhibited the capacity to revert the neurotoxicity induced by 6-OHDA to values higher or similar to the vitamin E (90.11 ± 3.74% of viable cells). The neuroprotective effects were mediated by the mitigation of reactive oxygen species (ROS) generation, mitochondrial dysfunctions and DNA damage, together with the reduction of Caspase-3 activity. Compounds belonging to different chemical classes, such as terpenes, alcohols, carboxylic acids, aldehydes, esters, ketones, saturated and unsaturated hydrocarbons were tentatively identified by GC-MS. The results show that  is a relevant source of neuroprotective agents, with particular interest for preventive therapeutics.

Natural Approaches for Neurological Disorders-The Neuroprotective Potential of .

The ever-increasing interest in keeping a young appearance and healthy skin has leveraged the skincare industry. This, coupled together with the increased concern regarding the safety of synthetic products, has boosted the demand for new and safer natural ingredients. Accordingly, the aim of this study was to evaluate the dermatological potential of the brown seaweed  The antioxidant, anti-enzymatic, antimicrobial, photoprotective and anti-inflammatory properties of five  fractions (F1-F5) were evaluated. The ethyl acetate fraction (F3) demonstrated the most promising results, with the best ability to scavenge 2,2-diphenyl-1-picrylhydrazyl (DPPH) radicals (EC of 140.1 µg/mL) and the capacity to reduce reactive oxygen species (ROS) production promoted by UVA and UVB radiation in 3T3 cells, revealing its antioxidant and photoprotective potential. This fraction also exhibited the highest anti-enzymatic capacity, inhibiting the activities of collagenase, elastase and tyrosinase (IC of 7.2, 4.8 and 85.9 µg/mL, respectively). Moreover, F3 showed anti-inflammatory potential, reducing TNF-α and IL-6 release induced by LPS treatment in RAW 264.7 cells. These bioactivities may be related to the presence of phenolic compounds, such as phlorotannins, as demonstrated by NMR analysis. The results highlight the potential of  as a source of bioactive ingredients for further dermatological applications.

Unravelling the Dermatological Potential of the Brown Seaweed .

Parkinsons Disease (PD) is the second most common neurodegenerative disease worldwide, and is characterized by a progressive degeneration of dopaminergic neurons. Without an effective treatment, it is crucial to find new therapeutic options to fight the neurodegenerative process, which may arise from marine resources. Accordingly, the goal of the present work was to evaluate the ability of the monoterpenoid lactone Loliolide, isolated from the green seaweed , to prevent neurological cell death mediated by the neurotoxin 6-hydroxydopamine (6-OHDA) on SH-SY5Y cells and their anti-inflammatory effects in RAW 264.7 macrophages. Loliolide was obtained from the diethyl ether extract, purified through column chromatography and identified by NMR spectroscopy. The neuroprotective effects were evaluated by the MTT method. Cells' exposure to 6-OHDA in the presence of Loliolide led to an increase of cells' viability in 40%, and this effect was mediated by mitochondrial protection, reduction of oxidative stress condition and apoptosis, and inhibition of the NF-kB pathway. Additionally, Loliolide also suppressed nitric oxide production and inhibited the production of TNF-α and IL-6 pro-inflammatory cytokines. The results suggest that Loliolide can inspire the development of new neuroprotective therapeutic agents and thus, more detailed studies should be considered to validate its pharmacological potential.

Loliolide, a New Therapeutic Option for Neurological Diseases? In Vitro Neuroprotective and Anti-Inflammatory Activities of a Monoterpenoid Lactone Isolated from .

The treatment of Parkinson´s disease (PD) has benefited from significant advances resulting from the increasing research efforts focused on new therapeutics. However, the current treatments for PD are mostly symptomatic, alleviating disease symptoms without reversing or retarding disease progression. Thus, it is critical to find new molecules that can result in more effective treatments. Within this framework, this study aims to evaluate the neuroprotective and anti-inflammatory effects of three compounds (eleganolone, eleganonal and fucosterol) isolated from the brown seaweed Bifurcaria bifurcata. In vitro neuroprotective effects were evaluated on a PD cellular model induced by the neurotoxin 6-hydroxydopamine (6-OHDA) on SH-SY5Y human cells, while lipopolysaccharide (LPS) - stimulated RAW 264.7 macrophages were used to evaluate the anti-inflammatory potential. Additionally, the underlying mechanisms of action were also investigated. Compounds were isolated by preparative chromatographic methods and their structural elucidation attained by NMR spectroscopy. Among the tested compounds, eleganolone (0.1-1 µM; 24 h) reverted the neurotoxicity induced by 6-OHDA in about 20%. The neuroprotective effects were mediated by mitochondrial protection, reduction of oxidative stress, inflammation and apoptosis, and inhibition of NF-kB pathway. The results suggest that eleganolone may provide advantages in the treatment of neurodegenerative conditions and, therefore, should be considered for future preclinical studies.

Disclosing the potential of eleganolone for Parkinson's disease therapeutics: Neuroprotective and anti-inflammatory activities.

Marine fungi and, particularly, endophytic species have been recognised as one of the most prolific sources of structurally new and diverse bioactive secondary metabolites with multiple biotechnological applications. Despite the increasing number of bioprospecting studies, very few have already evaluated the cosmeceutical potential of marine fungal compounds. Thus, this study focused on a frequent seaweed in the Portuguese coast, Halopteris scoparia, to identify the endophytic marine fungi associated with this host, and assess their ability to biosynthesise secondary metabolites with antioxidative, enzymatic inhibitory (hyaluronidase, collagenase, elastase and tyrosinase), anti-inflammatory, photoprotective, and antimicrobial (Cutibacterium acnes, Staphylococcus epidermidis and Malassezia furfur) activities. The results revealed eight fungal taxa included in the Ascomycota, and in the most representative taxonomic classes in marine ecosystems (Eurotiomycetes, Sordariomycetes and Dothideomycetes). These fungi were reported for the first time in Portugal and in association with H. scoparia, as far as it is known. The screening analyses showed that most of these endophytic fungi were producers of compounds with relevant biological activities, though those biosynthesised by Penicillium sect. Exilicaulis and Aspergillus chevalieri proved to be the most promising ones for being further exploited by dermocosmetic industry. The chemical analysis of the crude extract from an isolate of A. chevalieri revealed the presence of two bioactive compounds, echinulin and neoechinulin A, which might explain the high antioxidant and UV photoprotective capacities exhibited by the extract. These noteworthy results emphasised the importance of screening the secondary metabolites produced by these marine endophytic fungal strains for other potential bioactivities, and the relevance of investing more efforts in understanding the ecology of halo/osmotolerant fungi.

Marine endophytic fungi associated with Halopteris scoparia (Linnaeus) Sauvageau as producers of bioactive secondary metabolites with potential dermocosmetic application.

Resulting from the growing human population and the long dependency on fossil-based energies, the planet is facing a critical rise in global temperature, which is affecting all ecosystem networks. With a growing consciousness this issue, the EU has defined several strategies towards environment sustainability, where biodiversity restoration and preservation, pollution reduction, circular economy, and energetic transition are paramount issues. To achieve the ambitious goal of becoming climate-neutral by 2050, it is vital to mitigate the environmental footprint of the energetic transition, namely heavy metal pollution resulting from mining and processing of raw materials and from electronic waste disposal. Additionally, it is vital to find alternative materials to enhance the efficiency of energy storage devices. This review addresses the environmental challenges associated with energetic transition, with particular emphasis on the emergence of new alternative materials for the development of cleaner energy technologies and on the environmental impacts of mitigation strategies. We compile the most recent advances on natural sources, particularly seaweed, with regard to their use in metal recycling, bioremediation, and as valuable biomass to produce biochar for electrochemical applications.

Seaweed's Role in Energetic Transition-From Environmental Pollution Challenges to Enhanced Electrochemical Devices.

In recent decades, seaweeds have proven to be an excellent source of bioactive molecules. Presently, the seaweed  is harvested in a small area of the Portuguese coast exclusively for agar extraction. The aim of this work was to fully disclosure  as a sustainable source of antimicrobial ingredients for new dermatological formulations, highlighting its potential to be explored in a circular economy context. For this purpose, after a green sequential extraction, these seaweed fractions (F1-F5) were chemically characterized (H NMR) and evaluated for their antimicrobial potential against ,  and . The most active fractions were also evaluated for their effects on membrane potential, membrane integrity and DNA damage. Fractions F2 and F3 displayed the best results, with IC values of 16.1 (7.27-23.02) μg/mL and 51.04 (43.36-59.74) μg/mL against , respectively, and 53.29 (48.75-57.91) μg/mL and 102.80 (87.15-122.30) μg/mL against , respectively. The antimicrobial effects of both fractions seem to be related to membrane hyperpolarization and DNA damage. This dual mechanism of action may provide therapeutic advantages for the treatment of skin dysbiosis-related diseases.

Gelidiales Are Not Just Agar-Revealing the Antimicrobial Potential of  for Skin Disorders.

The growing knowledge about the harmful effects caused by some synthetic ingredients present in skincare products has led to an extensive search for natural bioactives. Thus, this study aimed to investigate the dermatological potential of five fractions (F1-F5), obtained by a sequential extraction procedure, from the brown seaweed . The antioxidant (DPPH, FRAP, ORAC and TPC), anti-enzymatic (collagenase, elastase, hyaluronidase and tyrosinase), antimicrobial (,  and ), anti-inflammatory (nitric oxide, tumor necrosis factor-α, interleukin-6 and interleukin-10) and photoprotective (reactive oxygen species) properties of all fractions were evaluated. The ethyl acetate fraction (F3) displayed the highest antioxidant and photoprotective capacity, reducing ROS levels in UVA/B-exposed 3T3 fibroblasts, and the highest anti-enzymatic capacity against tyrosinase (IC value: 89.1 µg/mL). The solid water-insoluble fraction (F5) revealed the greatest antimicrobial activity against  growth (IC value: 12.4 µg/mL). Furthermore, all fractions demonstrated anti-inflammatory potential, reducing TNF-α and IL-6 levels in RAW 264.7 macrophages induced with lipopolysaccharides. Chemical analysis of the  fractions by NMR revealed the presence of different classes of compounds, including lipids, polyphenols and sugars. The results highlight the potential of  to be incorporated into new nature-based skincare products.

-A Source of Natural Active Ingredients for Greener Skincare Formulations.

Gastrointestinal diseases, such as peptic ulcers, are caused by a damage in the gastric mucosa provoked by several factors. This stomach injury is regulated by many inflammatory mediators and is commonly treated with proton-pump inhibitors, histamine H2 receptor blockers and antacids. However, various medicinal plants have demonstrated positive effects on gastric ulcer treatment, including plants of the  genus. The aim of this study was to evaluate the antiulcer and anti-inflammatory activities of the stem bark ethanolic extract of  (A. St.-Hil.) Ravenna. We performed a preliminary quantification of phenolic compounds by high-performance liquid chromatography-diode array detection (HPLC-DAD), followed by the prospection of other chemical groups through nuclear magnetic resonance (NMR) spectroscopy. A set of in vitro assays was used to evaluate the extract potential regarding its antioxidant activity (DPPH: 19.83 ± 0.34 µg/mL; TPC: 307.20 ± 6.20 mg GAE/g of extract), effects on cell viability and on the release of TNF-α in whole human blood. Additionally, in vivo assays were performed to evaluate the leukocyte accumulation and total protein quantification in carrageenan-induced air pouch, as well as the antiulcerogenic effect of the extract on an ethanol-induced ulcer in rats. The extract contains flavonoids and phenolic compounds, as well as sugars and quinic acid derivatives exhibiting potent antioxidant activity and low toxicity. The extract reduced the release of TNF-α in human blood and inhibited the activity of p38α (1.66 µg/mL), JAK3 (5.25 µg/mL), and JNK3 (8.34 µg/mL). Moreover, it reduced the leukocyte recruitment on the pouch exudate and the formation of edema, reverting the effects caused by carrageenan. The extract presented a significant prevention of ulcer formation and a higher reduction than the reference drug, Omeprazole. Therefore,  extract has demonstrated relevant therapeutic potential for the treatment of gastric diseases, deserving the continuation of further studies to unveil the mechanisms of action of plant bioactive ingredients.

Antiulcerogenic Potential of the Ethanolic Extract of  (A. St.-Hil.) Ravenna Evaluated by In Vitro and In Vivo Studies.

The marine environment presents itself as a treasure chest, full of a vast diversity of organisms yet to be explored. Among these organisms, macroalgae stand out as a major source of natural products due to their nature as primary producers and relevance in the sustainability of marine ecosystems. Sulfated polysaccharides (SPs) are a group of polymers biosynthesized by macroalgae, making up part of their cell wall composition. Such compounds are characterized by the presence of sulfate groups and a great structural diversity among the different classes of macroalgae, providing interesting biotechnological and therapeutical applications. However, due to the high complexity of these macromolecules, their chemical characterization is a huge challenge, driving the use of complementary physicochemical techniques to achieve an accurate structural elucidation. This review compiles the reports (2016-2021) of state-of-the-art methodologies used in the chemical characterization of macroalgae SPs aiming to provide, in a simple way, a key tool for researchers focused on the structural elucidation of these important marine macromolecules.

Sulfated Polysaccharides from Macroalgae-A Simple Roadmap for Chemical Characterization.

The increase in the life expectancy average has led to a growing elderly population, thus leading to a prevalence of neurodegenerative disorders, such as Parkinson's disease (PD). PD is the second most common neurodegenerative disorder and is characterized by a progressive degeneration of the dopaminergic neurons in the substantia  (SNpc). The marine environment has proven to be a source of unique and diverse chemical structures with great therapeutic potential to be used in the treatment of several pathologies, including neurodegenerative impairments. This review is focused on compounds isolated from marine organisms with neuroprotective activities on in vitro and in vivo models based on their chemical structures, taxonomy, neuroprotective effects, and their possible mechanism of action in PD. About 60 compounds isolated from marine bacteria, fungi, mollusk, sea cucumber, seaweed, soft coral, sponge, and starfish with neuroprotective potential on PD therapy are reported. Peptides, alkaloids, quinones, terpenes, polysaccharides, polyphenols, lipids, pigments, and mycotoxins were isolated from those marine organisms. They can act in several PD hallmarks, reducing oxidative stress, preventing mitochondrial dysfunction, α-synuclein aggregation, and blocking inflammatory pathways through the inhibition translocation of NF-kB factor, reduction of human tumor necrosis factor α (TNF-α), and interleukin-6 (IL-6). This review gathers the marine natural products that have shown pharmacological activities acting on targets belonging to different intracellular signaling pathways related to PD development, which should be considered for future pre-clinical studies.

Marine-Derived Components: Can They Be a Potential Therapeutic Approach to Parkinson's Disease?

This work addresses the potential of the red seaweed  as a source of bioactive ingredients for skin health and wellness in response to the growing awareness regarding the significance of sustainable strategies in developing new nature-based dermocosmetic products. Hydroalcoholic extracts from the dried biomass were subjected to sequential liquid-liquid partitions, affording five different fractions (F1-F5). Their cosmetic potential was assessed through a set of in vitro assays concerning their antioxidant, photoprotective, and healing properties. Additionally, their cytotoxicity in HaCaT cells and their capacity to induce inflammation in RAW 264.7 cells were also evaluated. As a proof-of-concept, O/W emulsions were prepared, and emulsion stability was assessed by optical microscopy, droplet size analysis, centrifugation tests, and rheology analysis. Furthermore, in vivo tests were conducted with the final formulation to assess its antioxidant capacity. At subtoxic concentrations, the most lipophilic fraction has provided photoprotection against UV light-induced photooxidation in HaCaT cells. This was conducted together with the aqueous fraction, which also displayed healing capacities. Regarding the physical and stability assays, the best performance was achieved with the formulation containing 1% aqueous extract, which exhibited water retention and antioxidant properties in the in vivo assay. In summary,  displayed itself as a potential source of bioactive ingredients with multitarget properties for dermatological use.

Article	Year
Antioxidant and Neuroprotective Potential of the Brown Seaweed in an in vitro Parkinson's Disease Model. Marine drugs\| PubMed ID: 30717087	2019
Risk assessment of trihalomethanes exposure by consumption of IV gamma products: Evidences from a Portuguese regional survey. Toxicology reports\| PubMed ID: 32071882	2020
Natural Approaches for Neurological Disorders-The Neuroprotective Potential of . Molecules (Basel, Switzerland)\| PubMed ID: 33238492	2020
Unravelling the Dermatological Potential of the Brown Seaweed . Marine drugs\| PubMed ID: 33671016	2021
Loliolide, a New Therapeutic Option for Neurological Diseases? In Vitro Neuroprotective and Anti-Inflammatory Activities of a Monoterpenoid Lactone Isolated from . International journal of molecular sciences\| PubMed ID: 33672866	2021
Disclosing the potential of eleganolone for Parkinson's disease therapeutics: Neuroprotective and anti-inflammatory activities. Pharmacological research\| PubMed ID: 33812007	2021
Marine endophytic fungi associated with Halopteris scoparia (Linnaeus) Sauvageau as producers of bioactive secondary metabolites with potential dermocosmetic application. PloS one\| PubMed ID: 33983974	2021
Seaweed's Role in Energetic Transition-From Environmental Pollution Challenges to Enhanced Electrochemical Devices. Biology\| PubMed ID: 35336831	2022
Gelidiales Are Not Just Agar-Revealing the Antimicrobial Potential of for Skin Disorders. Antibiotics (Basel, Switzerland)\| PubMed ID: 35453232	2022
-A Source of Natural Active Ingredients for Greener Skincare Formulations. Molecules (Basel, Switzerland)\| PubMed ID: 36235032	2022
Antiulcerogenic Potential of the Ethanolic Extract of (A. St.-Hil.) Ravenna Evaluated by In Vitro and In Vivo Studies. International journal of molecular sciences\| PubMed ID: 36555275	2022
Sulfated Polysaccharides from Macroalgae-A Simple Roadmap for Chemical Characterization. Polymers\| PubMed ID: 36679279	2023
Marine-Derived Components: Can They Be a Potential Therapeutic Approach to Parkinson's Disease? Marine drugs\| PubMed ID: 37623732	2023
New Insights into the Dermocosmetic Potential of the Red Seaweed . Antioxidants (Basel, Switzerland)\| PubMed ID: 37759987	2023

Alice Martins

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