Search Results (107)

This study employed response surface methodology for the first-time optimization of the ultrasound-assisted extraction (UAE) of the total phenolic content (TPC) and ABTS from edible lichens, including Evernia divaricata, Evernia prunastri, Pseudevernia furfuracea, Bryoria fuscescens, and Lobaria pulmonaria. Fourteen experimental points were generated using Design Expert Software, with the extraction temperature (25–40 °C), extraction time (5–20 min), and ethanol concentration (0–80%) as independent variables, and TPC and ABTS as dependent variables. The phenolic profile and mineral and antibacterial properties of the optimized lichen extracts were determined. Evernic and usnic acid were found in Evernia species. Atranorin was detected only in P. furfuracea. Fumarprotocetraric acid was found exclusively in B. fuscescens and was not detected in any of the other lichens. Calcium was found to have the highest mineral content in all the lichens, followed by potassium. L. pulmonaria, showing the lowest inhibition effect against all tested bacteria, while E. divaricata exhibited the most effective inhibition. Full article

Chirality plays a key role in the effectiveness and toxicity of bioactive compounds. Usnic acid (UA), a lichen metabolite, exists as two enantiomers. Despite numerous studies on its biological properties, enantioselective aspects remain poorly recognized. This study assessed the cytotoxicity of UA enantiomers against colon, prostate, thyroid, brain, and breast cancer cell lines, as well as non-cancerous cells. Cell viability was determined by the MTT assay after 24, 48, and 72 h. Colon cancer HCT116 cells were the most sensitive (IC₅₀ ~10 µg/mL, 72 h), with no enantiomeric dominance. In prostate cancer PC3 cells, (+)-UA was more effective. Moderate cytotoxic effect was noted for thyroid cancer cells; however, this was evaluated for the first time. MDA-MB-231 breast cancer cells were strongly affected (IC₅₀ 15.8 and 20.2 µg/mL for (+)- and (−)-UA, 72 h), as compared to MCF7 cells. Brain cancer cells were the least affected, as so were normal astrocytes. UA had no effect on normal colon epithelial cells but showed moderate toxicity in prostate, thyroid, and breast cells. To conclude, the overall cytotoxicity of (+)-UA was stronger than its (−)-enantiomer, while the latter compound was more toxic to normal cells. These findings highlight the advantage of (+)-UA, especially in chemopreventive strategies. Full article

Root and crown rot in strawberries caused by Neopestalotiopsis rosae (N. rosae) results in yield losses of approximately 70%. The main method of control is based on the application of fungicides; however, the excessive use of these products can induce resistance by pathogens to the active ingredients. The use of secondary metabolites is an alternative to disease management. Usnic acid (UA), a secondary metabolite produced by lichens, has shown antimicrobial and antifungal activities that could be useful for the management of phytopathogens, particularly the (+) enantiomer. To provide alternatives to fungicides, the potential of UA as an alternative for N. rosae management was evaluated under in vitro and in vivo conditions. Using the “poisoned medium” technique, concentrations of 0 (UA0), 100 (UA1), 200 (UA2), and 400 (UA4) µg/mL UA at a dose of 2.5 mL/L PDA were evaluated on N. rosae mycelial growth and the number of spores. The UA at 400 µg/mL exhibited a fungistatic effect, reducing the mycelial growth of isolates of N. rosae in 50–60%. In the in vivo assay, sprayed UA (400 µg/mL) reduced hydrogen peroxide (48.59%) and malonaldehyde (77.62%) contents in “Albion” strawberry seedlings inoculated with 466 and FREC2 strains, respectively. These findings suggest that UA could be a potential tool for N. rosae management and could help mitigate the oxidative stress induced by infection. However, field trials are required to evaluate and validate this response. Full article

In this study, a global response analysis was performed to explore the mechanism of action of Usnic acid and its synergy with Norfloxacin, a well-known quinolone antibiotic to which MRSA clinical isolates showed resistance (MIC, 500 µg/mL). A microdilution assay, a growth kinetics analysis, a microscopic analysis, and cell-based assays consistently showed that Usnic acid possesses strong anti-staphylococcal activity (MIC, 7.8 µg/mL), causes cell leakage, modulates efflux pump activity, and synergizes with Norfloxacin against the multi-drug-resistant clinical isolate MRSA 2071. Whole-cell proteome profiling using gel-free proteomics-based nano-LC-ESI-QTOF-MS/MS revealed several proteins whose expression was significantly modulated by Usnic acid and Norfloxacin alone or in combination. Usnic acid downregulated the abundance of RNA polymerase subunits (RpoB and RpoC), carbamoyl phosphate synthase large subunit (PyrAB), chaperone (GroEL), and adenylosuccinate synthetase (PurA). Interestingly, proteins found to be upregulated in the presence of Usnic acid and Norfloxacin included oxidative-stress-related proteins such as peroxidase (Tpx), alkyl hydroperoxide reductase (AphC), and general stress protein (UspA). This study clearly shows that Usnic acid affects numerous cellular targets and can potentiate the action of Norfloxacin. Furthermore, an in vivo study showed that UA at low concentrations prevents body weight gain, but changes in other tested toxicological parameters were found to be within normal limits. Thus, UA at low doses appears to be a promising candidate for repurposing old antibiotics through combination therapy against MRSA infections. Full article

Lichen-specific natural products exhibit a wide range of biological activities, which makes them potentially useful in the pharmaceutical, cosmetic, and nutritional industries. In the present study, a capillary electrophoresis method was developed and optimized for the separation of seven major metabolites, physodic acid, 3-hydroxyphysodic acid, atranorin, physodalic acid, chloroatranorin, salazinic acid, and protocetraric acid, found in Hypogymnia physodes. The optimization was performed using a design of experiments approach, focusing on four critical parameters: boric acid concentration, deoxycholic acid concentration, methanol content, and buffer pH. The overall separation efficiency was used as the response factor for optimization. The optimal separation conditions were achieved using a buffer composed of 60 mM boric acid, 70 mM deoxycholic acid, and 14% methanol at pH 9.6. The validated method was subsequently applied for the chemophenetic analysis of 28 lichen species belonging to the families Cladoniaceae, Parmeliaceae, Physciaceae, Ramalinaceae, and Teloschistaceae. In addition to the above-mentioned lichen compounds, the lichens examined showed the presence of evernic acid, usnic acid, and physicon. The developed CE method offers a reliable and efficient tool for the characterization of lichen metabolites, with potential applications in both botany and natural product research. Full article

Usnetic acid is a dibenzofuran-2-ylacetic acid that can be obtained by alkaline degradation of a secondary lichen metabolite—usnic acid. In the present paper, the product of the reaction of usnetic acid with a mild fluorinating agent, Selectfluor^®, was obtained. The structure of the product was proved by a set of physical methods, including ¹H, ¹³C, ¹⁹F NMR, HMBC, HSQC, HRMS and IR spectroscopy. Full article

Background: Disruption of the natural balance of the skin microbiota can impair wound healing and contribute to chronic infections. Identifying the bacterial species involved and understanding their antimicrobial susceptibility profiles are essential for guiding treatment, especially given the growing threat of antibiotic resistance. Methods: This study characterized the virulence and antibiotic resistance phenotypes of 43 bacterial strains isolated from chronic wounds. The antimicrobial activity of selected essential oils (sandalwood, ylang-ylang, sage, cajeput, and juniper), pharmaceutical products (propolis tinctures, usnic acid), and probiotic lactic acid bacteria strains (Lactobacillus spp., Lactococcus lactis) was assessed using qualitative and quantitative assays, including MIC, MICBA, and co-culture evaluations. Results: Gram-positive strains were more sensitive to essential oils than Gram-negative strains, with sandalwood, ylang-ylang, and propolis tincture showing the strongest antibacterial effects. These agents also showed significant biofilm inhibition. Probiotic strains exhibited antimicrobial activity against Staphylococcus aureus and Morganella morganii, with Lactobacillus paracasei and Lactobacillus rhamnosus being particularly effective in reducing bacterial growth and adhesion in vitro. Conclusions: Essential oils and probiotic strains demonstrate promising antimicrobial effects against chronic wound pathogens and may serve as alternative or adjunctive treatments to antibiotics. Further clinical research and standardization are necessary to establish their safety, efficacy, and optimal application protocols. Full article

In a hidden corner of the Earth, an ongoing war is being waged: a battle between lichens and microorganisms. Lichens, ancient and unique symbiotic organisms, with their unique survival wisdom, are bursting with vitality in extreme environments. Over 80% of secondary metabolites in lichens are not found in other organisms, making lichen-derived compounds a promising resource for the development of new drugs, particularly against drug-resistant microorganisms, due to their distinctive chemical structures and biological activities. This article aims to explore in depth the lichen species exhibiting antimicrobial activity and their antimicrobial metabolites and focus on unique compounds such as divaricatic acid, usnic acid, vulpinic acid, salazinic acid, and rhizocarpic acid, which demonstrate significant antimicrobial effects against various resistant microorganisms, including methicillin-resistant Staphylococcus aureus, drug-resistant Mycobacterium tuberculosis, and Candida albicans and other drug-resistant microorganisms. Meanwhile, this paper discusses the potential applications and challenges associated with the use of lichens in medicine, agriculture, and food industry, aiming to elucidate these mysterious organisms for lichen researchers and enthusiasts while promoting further research and applications in the field of antimicrobials. Full article

Background/Objectives: Titanium dental implants, while highly successful, face challenges due to polymicrobial infections leading to peri-implantitis and implant failure. Biofilm formation on implant surfaces is the primary cause of these infections, with factors such as matrix production and cross-kingdom interactions contributing to the microbial accumulation of bacterial and fungal pathogens species. To combat this issue, naturally derived molecules have been reported to overcome the hurdle of antimicrobial resistance against the application of conventional antibiotics and antifungals. Methods: The present study aimed to employ the lichen-derived molecules, usnic acid (UA), to retard the development of biofilms of bacterial and fungal pathogens on the surface of titanium kept in the human artificial saliva (HAS) working as a growth-supporting, host-mimicking media. Results: The minimum inhibitory concentration of UA in HAS towards Candida albicans was >512 µg/mL, whereas against Staphylococcus aureus and Streptococcus mutans, it was determined to be 512 µg/mL. Whereas, in the standard growth media, the MIC value of UA towards S. mutans and S. aureus were 8 and 16 µg/mL; however, against C. albicans, it was 512 µg/mL. UA synergistically enhanced the efficacy of the antibiotics toward bacterial pathogens and the efficacy of antifungals against C. albicans. The antibiofilm results depict the fact that in the HAS, UA significantly reduced both mono-species of S. mutans, S. aureus, and C. albicans and mixed-species biofilm of C. albicans with S. mutans and S. aureus on the surface of the titanium. Conclusions: The present study showed that UA is a promising natural drug that can control oral polymicrobial disease as a result of the application of dental implants. Full article

Tyrosyl-DNA phosphodiesterases 1 and 2 (TDP1 and TDP2) are important DNA repair enzymes that remove various adducts from the 3′- and 5′-ends of DNA, respectively. The suppression of the activity of these enzymes is considered as a promising adjuvant therapy for oncological diseases in combination with topoisomerase inhibitors. The simultaneous inhibition of TDP1 and TDP2 may result in greater antitumor effects, as these enzymes can mimic each other’s functions. We have previously shown that usnic acid-based sulfides can act as dual inhibitors, with TDP1 activity in the low micromolar range and their TDP2 at 1 mM. The oxidation of their sulfide moieties to sulfoxides led to an order of magnitude decrease in their cytotoxicity potential, while their TDP1 and TDP2 activity was preserved. In this work, we synthesized new series of usnic acid-based sulfides and their oxidized analogues, i.e., sulfoxides and sulfones, to systematically study these irregularities. The new compounds inhibit TDP1 with IC₅₀ values (the concentration of inhibitor required to reduce enzyme activity by half) in the 0.33–25 μM range. Most sulfides and some sulfoxides and sulfones inhibit TDP2 with an IC₅₀ = 138−421 μM. In addition, the most active compounds synergized (×4) with topotecan on the HeLa cell line as well as causing dose-dependent DNA damage, as confirmed by Comet assay. Sulfides with the 6-methylbenzoimidazol-2-yl substituent (8f, IC₅₀ = 0.33/138 μM, TDP1/2) and sulfones containing a pyridine-2-yl fragment (12k, IC₅₀ = 2/228 μM, TDP1/2) are the most potent derivatives and, therefore, are promising for further development. Full article

Cationic antimicrobial peptides (AMPs), also called host defence peptides, have established antimicrobial and anticancer activities. Conjugation of an AMP to a bioactive molecule with complementary activity can address some of the clinical limitations of the peptide candidate. This approach has been particularly applied in antimicrobial applications of AMPs, but it remains relatively less explored in the generation of anticancer candidates. In this study, two usnic acid derivatives, based on hydrazinothiazole and benzylidenefuranone pharmacophore moieties, respectively, were conjugated to L-K6, a lysine/leucine-rich AMP, through a new pyrazole ligation intrinsically driven by the cargo molecule. Both components, the usnic acid derivative and the peptide, are selectively active against cancer cells, by targeting the human DNA repair enzyme tyrosyl-DNA phosphodiesterase 1 (TDP1) and through DNA damage, respectively. The two conjugates, based on a hydrazone linkage, exhibited pleiotropic effects, ranging from reduction in the activity of the parent drugs to their conservation or even enhancement. Notably, the conjugates retained some anti-TDP1 activity and displayed intermediate, or even higher, cytotoxicities against glioblastoma cells, compared to their individual components. Full article

In the present work, the phytotoxic effects of secondary metabolites extracted from lichen Ramalina celastri (usnic acid) and lichen Stereocaulon ramulosum (a naturally occurring mixture of atranorin and perlatolic acid, approx. 3:1) on cultures of the aposymbiotically grown lichen photobiont Asterochloris erici were evaluated. Algae were cultivated on the surface of glass microfiber disks with applied crystals of lichen extracts for 14 days. The toxicity of each extract was tested at the two selected doses in quantities of 0.01 mg/disk and 0.1 mg/disk. Cytotoxicity of lichen extracts was assessed using selected physiological parameters, such as growth (biomass production) of photobiont cultures, content of soluble proteins, chlorophyll a fluorescence, chlorophyll a integrity, contents of chlorophylls and total carotenoids, hydrogen peroxide, superoxide anion, TBARS, ascorbic acid (AsA), reduced (GSH) and oxidized (GSSG) glutathione, and composition of selected organic acids of the Krebs cycle. The application of both tested metabolic extracts decreased the growth of photobiont cells in a dose-dependent manner; however, a mixture of atranorin and perlatolic acid was more effective when compared to usnic acid at the same dose tested. A higher degree of cytotoxicity of extracts from lichen S. ramulosum when compared to identical doses of extracts from lichen R. celastri was also confirmed by a more pronounced decrease in chlorophyll a fluorescence and chlorophyll a integrity, decreased content of chlorophylls and total carotenoids, increased production of hydrogen peroxide and superoxide anion, peroxidation of membrane lipids (assessed as TBARS), and a strong decrease in non-enzymatic antioxidants such as AsA, GSH, and GSSG. The cytotoxicity of lichen compounds was confirmed by a strong alteration in the composition of selected organic acids included in the Krebs cycle. The increased ratio between pyruvic acid and citric acid was a very sensitive parameter of phytotoxicity of lichen secondary metabolites to the algal partner of symbiosis. Secondary metabolites of lichens are potent allelochemicals and play significant roles in maintaining the balance between mycobionts and photobionts, forming lichen thallus. Full article

Cancer, a complex group of diseases marked by uncontrolled cell growth and invasive behavior, is characterized by distinct hallmarks acquired during tumor development. These hallmarks, first proposed by Douglas Hanahan and Robert Weinberg in 2000, provide a framework for understanding cancer’s complexity. Targeting them is a key strategy in cancer therapy. It includes inhibiting abnormal signaling, reactivating growth suppressors, preventing invasion and metastasis, inhibiting angiogenesis, limiting replicative immortality, modulating the immune system, inducing apoptosis, addressing genome instability and regulating cellular energetics. Usnic acid (UA) is a natural compound found in lichens that has been explored as a cytotoxic agent against cancer cells of different origins. Although the exact mechanisms remain incompletely understood, UA presents a promising compound for therapeutic intervention. Understanding its impact on cancer hallmarks provides valuable insights into the potential of UA in developing targeted and multifaceted cancer therapies. This article explores UA activity in the context of disrupting hallmarks in cancer cells of different origins based on recent articles that emphasize the molecular mechanisms of this activity. Full article

The genus Xanthoparmelia includes several subcrustose, squamulose, small foliose, and small subfruticose species, primarily in the Southern Hemisphere. Here, we report on the first small foliose species lacking usnic acid in the genus occurring in the Holarctic. The species has been previously known as Lecanora olivascens Nyl., but subsequent studies of the morphology, secondary chemistry, and molecular data of the nuITS rDNA indicate that this species instead belongs to Xanthoparmelia. Consequently, the new combination Xanthoparmelia olivascens (Nyl.) V.J. Rico and G. Amo is proposed, and an epitype is designated here. We discuss the unique presence of a subcrustose Xanthoparmelia species lacking cortical usnic acid in the Northern Hemisphere. This species fits phylogenetically into a clade that was previously only known from the Southern Hemisphere, and hence represents another example of N-S disjunction in lichenized fungi. Full article