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- Cytotoxity induced by extracts of Pisolithustinctorius spores on human cancer and normal cell lines – evaluation of the anticancer potentialPublication . Alves, Ricardo; Preto, Marco; Oliveira, Rui S.; Martins, RosárioCancer is one of the most important causes of death and morbidity worldwide. According to the World Health Organization 8.2 million people have died from cancer in 2012. The treatment of this fast growing disease is regarded as a major challenge in health for the XXI century. Conventional anti-cancer therapy is usually aggressive to the patients due to systemic toxicity caused by lack of specificity of the chemotherapy drugs (Kidd, 2000). This has triggered the interest of the scientific community to search for new and more effective therapies, less aggressive than conventional treatments. Used since antiquity in oriental civilizations, fungi, in particular mushrooms have been shown to limit or reverse tumor growth (Kidd, 2000). Pisolithustinctorius (Basidiomycota) is a soil fungus with world-wide distribution that it known to form ectomycorrhizal symbioses in forest ecosystems (Kope & Fortin, 1990). The sporocarps of this fungus contain Pisosterol, a triterpene that has been shown to have antitumor activity against some cancer cell lines (Montenegro et al., 2004). Nevertheless, no studies have focused on the anticancer potential of other structures such as spores and thus the anticancer potential of P. tinctorius , remains largely unknown.
- Toxicity assessment of crude and partially purified extracts of marine Synechocystis and Synechococcus cyanobacterial strains in marine invertebratesPublication . Martins, Rosario; Fernandez, Nuria; Beiras, Ricardo; Vasconcelos, VítorAmong the Cyanoprokaryota, the genera Synechocystis and Synechococcus have rarely been studied with respect to potential toxicity. This is particularly true with marine environments where studies about the toxicity of cyanobacteria are restricted to filamentous forms at the warmer temperate and tropical regions and also to filamentous forms at cold seas such as the Baltic Sea. In this study, we describe the effects of cyanobacterial strains of the Synechocystis and Synechococcus genera isolated from the marine coast of Portugal, on marine invertebrates. Crude and partially purified extracts at a concentration of 100 mg/ml of freeze-dried material of the marine strains were tested for acute toxicity in nauplii of the brine shrimp Artemia salina, in the rotifer Brachionus plicatillis and in embryos of the sea urchin Paracentrotus lividus and the mussel Mytilus galloprovincialis. The cyanobacterial extracts, especially the crude extract, had an impact on A. salina nauplii. No significant toxic effects were registered against the rotifer. A negative impact of all strains was recorded on the embryonic development of the sea urchin, with toxic effects resulting in an inhibition of embryogenesis or development of smaller larvae. To the mussel embryos, the effects of cyanobacterial extracts resulted in a complete inhibition of embryogenesis. The results of all assays indicate that Synechocystis and Synechococcus marine strains contained toxic compounds to marine invertebrates.
- Cyanobacteria for skin care and cosmeceutical formulationsPublication . Melo, Luísa; Morone, Janaína; Silva, Margarida; Ferraz, Ricardo; Martins, RosárioSkin acts as a barrier from external stimulus such as pollutants and ultraviolet radiation. Disturbs in the skin are involved in skin aging, which mostly results on a thinner epidermis, dryness, wrinkles, and loss of elasticity. Synthetic ingredients in cosmetics are known to be more toxic and to cause negative impact on the environment. In a society increasingly worried with human and environment health, natural skin care products gain special attention and boosted the search for natural sources. Due to the production of bioactive compounds, cyanobacteria emerged as an excellent source for cosmetic ingredients. Cyanobacteria strains of CIIMAR culture collection (LEGE-CC) have already proven to be interesting for cosmetic purposes [1,2]. In this project we aimed to evaluate the potential of four LEGE-CC cyanobacteria strains for skin care purposes. The cytotoxicity of acetonic and aqueous extracts was evaluated in keratinocytes (HaCAT), fibroblasts (3T3L1) and endothelial cells (hCMEC/D3) along with the total phenolic content and antioxidant properties. Cytotoxic assays revealed toxicity of acetonic extracts to keratinocytes in the higher concentrations tested. Aqueous extracts revealed low or non-cytotoxicity. Mainly in low concentrations of extracts, cellular proliferation was registered. Aqueous extracts from strain JM/RS021A and acetone extracts from strain JM/RS035B revealed higher values for total phenolic compounds. The acetone extracts of strain JM/RS021A showed the greatest scavenging activity values on the DPPH assay.
- Stalling the course of neurodegenerative diseases: could cyanobacteria constitute a new approach toward therapy?Publication . Ramos, Vitória; Reis, Mariana; Ferreira, Leonor; Silva, Ana Margarida; Ferraz, Ricardo; Vieira, Mónica; Vasconcelos, Vitor; Martins, RosárioNeurodegenerative diseases (NDs) are characterized by progressive and irreversible neuronal loss, accompanied by a range of pathological pathways, including aberrant protein aggregation, altered energy metabolism, excitotoxicity, inflammation, and oxidative stress. Some of the most common NDs include Alzheimer’s Disease (AD), Parkinson’s Disease (PD), Multiple Sclerosis (MS), Amyotrophic Lateral Sclerosis (ALS), and Huntington’s Disease (HD). There are currently no available cures; there are only therapeutic approaches that ameliorate the progression of symptoms, which makes the search for new drugs and therapeutic targets a constant battle. Cyanobacteria are ancient prokaryotic oxygenic phototrophs whose long evolutionary history has resulted in the production of a plethora of biomedically relevant compounds with anti-inflammatory, antioxidant, immunomodulatory, and neuroprotective properties, that can be valuable in this field. This review summarizes the major NDs and their pathophysiology, with a focus on the anti-neurodegenerative properties of cyanobacterial compounds and their main effects.
- Evaluation of the potential of marine cyanobacteria towards Alzheimer’s diseasePublication . Moreira, Ana Rita; Silva, Ana Margarida; Vieira, Mónica; Martins, RosárioAlzheimer’s disease (AD) is one of the most prevalent neurodegenerative diseases (NDs) and the major cause of dementia. Despite the constant efforts to achieve cure, no treatment is currently effective and new approaches are urgently needed. Marine biotechnology has been at the frontline for the bioprospection of natural sources to treat ND. Several marine compounds have already provided evidence for anti-AD drugs by acting in mechanisms such as the inhibition of acetylcholinesterase (AChE). Among marine organisms, cyanobacteria have been recognized as a major source of bioactive compounds and the potential towards AD has been highlighted, namely AChE inhibitors. This project is included in a screening program that aims to evaluate the potential of cyanobacteria from the Culture Collection of CIIMAR -LEGE-CC against AD. In this work, 80 fractions of marine cyanobacteria were screened for the potential to inhibit AChE since this is a proven enzyme target to ameliorate the loss of cholinergic signal and to induce cytotoxicity in the neuroblastoma cell line SHY5Y, in the normal fibroblasts cell line 3T3L1, and in the endothelial cells from central nervous system capillaries hCMEC/D3. Results considering the enzymatic assays and cytotoxic assays revealed that the fractions are not toxic or with low toxicity. The strains Phormidium sp; an unidentified filamentous Synechococcales and Nodosilinea nodulosa revealed moderate toxicity towards AChE.
- Hierridin B Isolated from a Marine Cyanobacterium Alters VDAC1, Mitochondrial Activity, and Cell Cycle Genes on HT-29 Colon Adenocarcinoma CellsPublication . Freitas, Sara; Martins, Rosario; Costa, Margarida; Leão, Pedro; Vitorino, Rui; Vasconcelos, Vitor; Urbatzka, RalphHierridin B was isolated from a marine cyanobacterium Cyanobium sp. strain and induced cytotoxicity selectively in HT-29 adenocarcinoma cells. The underlying molecular mechanism was not yet elucidated.
- Carotenoids from cyanobacteria modulate iNOS and inhibit the production of inflammatory mediators: Promising agents for the treatment of inflammatory conditionsPublication . Morone, Janaína; Vasconcelos, Vitor; Hentschke, Guilherme; Rosário Martins, Maria; Pinto, Eugénia; Lopes, GracilianaCyanobacteria are green multiproduct refineries of increasing interest for different industrial prospects. In this work, eleven cyanobacteria strains isolated from the Cape Verde archipelago were explored for their biotechnological applications in the field of inflammation. A biorefinery approach was employed to produce carotenoidtargeted extracts, further profiled by HPLC-PDA and explored for their ability to i) scavenge important physiological free radicals of oxygen (superoxide anion radical, O 2 •) and nitrogen (nitric oxide, • NO) involved in the inflammatory process ii) slow-down post-inflammatory hyperpigmentation and iii) modulate the activity of inf lammatory cytokine-producing enzymes, in enzymatic and cell systems comprising RAW 264.7 cells. The studied strains turned out to be important carotenoid producers (70.47–186.71 μ g mg 1 dry extract), mainly represented by β-carotene and zeaxanthin. The targeted-extracts stood-out for their potential to slow-down the inflammatory process through a multitarget approach: scavenging • NOandO 2 •, reducing inflammatory cytokines production through lipoxygenase inhibition, and modulating the inducible nitric oxide synthase in LPSstimulated RAW 264.7 cells, with strains of the order Nodosilineales revealing to be worth of further biotechnological exploitation.
- Structure of Hierridin C, Synthesis of Hierridins B and C, and Evidence for Prevalent Alkylresorcinol Biosynthesis in PicocyanobacteriaPublication . Costa, Margarida; Sampaio-Dias, Ivo E.; Castelo-Branco, Raquel; Scharfenstein, Hugo; Rezende de Castro, Roberta; Silva, Artur; Schneider, Maria Paula C.; Araújo, Maria João; Martins, Rosario; Domingues, Valentina F.; Nogueira, Fátima; Camões, Vera; Vasconcelos, Vitor M.; Leão, Pedro N.Small, single-celled planktonic cyanobacteria are ubiquitous in the world's oceans yet tend not to be perceived as secondary metabolite-rich organisms. Here we report the isolation and structure elucidation of hierridin C, a minor metabolite obtained from the cultured picocyanobacterium Cyanobium sp. LEGE 06113. We describe a simple, straightforward synthetic route to the scarcely produced hierridins that relies on a key regioselective halogenation step. In addition, we show that these compounds originate from a type III PKS pathway and that similar biosynthetic gene clusters are found in a variety of bacterial genomes, most notably those of the globally distributed picocyanobacteria genera Prochlorococcus, Cyanobium and Synechococcus.
- Exploring the bioactive potential of Pisolithus (Basidiomycota): comprehensive insights into antimicrobial, anticancer, and antioxidant properties for innovative applicationsPublication . S. Oliveira, Rui; Preto, Marco; Santos, Germana; Silva, Ana Margarida; Vasconcelos, Vitor; Martins, RosárioAddressing pressing health concerns, modern medical research seeks to identify new antimicrobials to combat drug resistance, novel molecules for cancer treatment, and antioxidants for inflammation-related diseases. Pisolithus (Basidiomycota) is a ubiquitous and widely distributed fungal genus in forest ecosystems, known for establishing ectomycorrhizal associations with a range of host plants, enhancing their growth, and conferring protection against biotic and abiotic stresses. Beyond ecological applications, Pisolithus yields bioactive compounds with medicinal potential. This comprehensive review explores the transversal biological activity of Pisolithus fungi, aiming to provide a thorough overview of their antimicrobial, anticancer, and antioxidant potential. The focus is on elucidating bioactive compounds within Pisolithus to trigger further research for innovative applications. Compounds from Pisolithus displayed antimicrobial activity against a broad spectrum of microorganisms, including antibiotic-resistant bacteria. The efficacy of Pisolithus-derived compounds matched established medications, emphasizing their therapeutic potential. In anticancer research, the triterpene pisosterol stood out with documented cytotoxicity against various cancer cell lines, showcasing promise for novel anticancer therapies. Pisolithus was also recognized as a potential source of antioxidants, with basidiocarps exhibiting high antioxidant activity. In vivo validation and comprehensive studies on a broader range of compounds, together with mechanistic insights into the mode of action of Pisolithus-derived compounds, are compelling areas for future research.
- Cyanobacteria secondary metabolites as biotechnological ingredients in natural anti-aging cosmetics: potential to overcome hyperpigmentation, loss of skin density and UV radiation-deleterious effectsPublication . Favas, Rita; Morone, Janaína; Martins, Rosário; Vasconcelos, Vitor; Lopes, GracilianaThe loss of density and elasticity, the appearance of wrinkles and hyperpigmentation are among the first noticeable signs of skin aging. Beyond UV radiation and oxidative stress, matrix metalloproteinases (MMPs) assume a preponderant role in the process, since their deregulation results in the degradation of most extracellular matrix components. In this survey, four cyanobacteria strains were explored for their capacity to produce secondary metabolites with biotechnological potential for use in anti-aging formulations. Leptolyngbya boryana LEGE 15486 and Cephalothrix lacustris LEGE 15493 from freshwater ecosystems, and Leptolyngbya cf. ectocarpi LEGE 11479 and Nodosilinea nodulosa LEGE 06104 from marine habitats were sequentially extracted with acetone and water, and extracts were analyzed for their toxicity in cell lines with key roles in the skin context (HaCAT, 3T3L1, and hCMEC). The non-toxic extracts were chemically characterized in terms of proteins, carotenoids, phenols, and chlorophyll a, and their anti-aging potential was explored through their ability to scavenge the physiological free radical superoxide anion radical (O2•−), to reduce the activity of the MMPs elastase and hyaluronidase, to inhibit tyrosinase and thus avoid melanin production, and to block UV-B radiation (sun protection factor, SPF). Leptolyngbya species stood out for anti-aging purposes: L. boryana LEGE 15486 presented a remarkable SPF of 19 (at 200 µg/mL), being among the best species regarding O2•− scavenging, (IC50 = 99.50 µg/mL) and also being able to inhibit tyrosinase (IC25 = 784 µg/mL), proving to be promising against UV-induced skin-aging; L. ectocarpi LEGE 11479 was more efficient in inhibiting MMPs (hyaluronidase, IC50 = 863 µg/mL; elastase, IC50 = 391 µg/mL), thus being the choice to retard dermal density loss. Principal component analysis (PCA) of the data allowed the grouping of extracts into three groups, according to their chemical composition; the correlation of carotenoids and chlorophyll a with MMPs activity (p < 0.01), O2•− scavenging with phenolic compounds (p < 0.01), and phycocyanin and allophycocyanin with SPF, pointing to these compounds in particular as responsible for UV-B blockage. This original survey explores, for the first time, the biotechnological potential of these cyanobacteria strains in the field of skin aging, demonstrating the promising, innovative, and multifactorial nature of these microorganisms.