Natural products include a diverse set of compounds of drug discovery that are currently being actively used to target tumor angiogenesis. In the present study, we evaluated the anti-angiogenic activities of secondary metabolite usnic acid isolated from Usena antarctica. We investigated the in vitro effects on proliferation, migration, and tube formation of VEGF- and bFGF-stimulated HUVECs. Ex ovo anti-angiogenic activity was evaluated using the CAM assay. Our findings demonstrated that usnic acid in the concentration of 33.57 µM inhibited VEGF (25 ng/mL) and bFGF (30 ng/mL)-induced HUVECs proliferation, migration, and tube formation. The ex ovo CAM model was used to confirm the results obtained from in vitro studies. VEGF- and bFGF-induced vessel formation was inhibited by usnic acid after 72 h in over 2-fold higher concentrations compared to in vitro. Subsequently, histological sections of affected chorioallantoic membranes were stained with hematoxylin–eosin and alcian blue to determine the number and diameter of vessels as well as the thickness of the individual CAM layers (ectoderm, mesoderm, endoderm). Usnic acid was able to suppress the formation of VEGF- and bFGF-induced vessels with a diameter of less than 100 μm, which was demonstrated by the reduction of mesoderm thickness as well.

Elemental composition of Vaccinium myrtillus L. and Vaccinium vitis-idaea L. has not been determined yet in detail. In our study, a scanning electron microscope coupled with an energy dispersive X-ray analyser (SEM–EDX) ensured the determination of 15 elements in the leaves of Vaccinium sp. growing in the control and the mine polluted locality. The soil elemental analyses showed a higher content of 11 elements from all determined in the mine-influenced samples. Elemental analyses of the control leaves showed the highest contents of all determined elements for V. vitis-idaea, except for carbon. The impact of pollution on V. myrtillus leaves caused significant increase in oxygen, natrium, magnesium, sulphur, chlorine, potassium and calcium content. The contents of carbon, nitrogen and silicon decreased substantially. In the case of the V. vitis-idaea leaves, the content of most of elements reduced, and in the case of six elements, even significantly. A significant increase was recorded for carbon, iron and copper. The effect of the environment has influenced the production of phytochemicals, assessed by HPLC–DAD. The production of flavonoids (hyperoside, isoquercetin) increased significantly in V. myrtillus and decreased to a detectable minimum in V. vitis-idaea. In the case of chlorogenic acid, arbutin and hydroquinone, their levels changed minimally in V. myrtillus, but significantly more in the V. vitis-idaea leaves. The importance of elemental analyses lies in detecting the presence of toxic elements or their harmful concentrations in plants that are the source of food or dietary supplements.

Infections caused by Malassezia (M.) pachydermatis in dogs are mostly treated with azole antifungals. Excessive use of these drugs is usually associated with an increased incidence of resistant isolates, which can be prevented by combining commonly used antifungals with natural bioactive compounds. The present study aimed at testing the effectiveness of a combination of selected azole derivatives showing low antifungal activity against M. pachydermatis isolates, with plant essential oil components displaying the highest efficacy. Among the four azole antifungals tested (itraconazole, posaconazole, clotrimazole, and miconazole), clotrimazole (a mean MIC of 7.62 μg/mL at 72 h and 7.24 μg/mL at 96 h) and miconazole (a mean MIC of 1.71 μg/mL at 72 h and 2.33 μg/mL at 96 h) exhibited the lowest antifungal efficacy. Out of the four plant essential oil components tested (eugenol, terpinene-4-ol, geraniol, and limonene), eugenol (an average MIC of 378.57 μg/mL at 72 h and 1180 μg/mL at 92 h) showed the highest antifungal activity. The checkerboard method was used to assess the interaction of these agents. The fractional inhibitory concentration index (FICI) values for the combination of clotrimazole with eugenol reached 1.43 at 72 h and 0.70 at 96 h and for the combination of miconazole with eugenol, 1.30 at 72 h and 0.45 at 96 h. A higher effect of the combinations was recorded at 96 h, when the combination of clotrimazole with eugenol showed an additive effect in 66.67% of the isolates, and the combination of miconazole and eugenol brought a synergistic effect in 57.14% of the isolates. The obtained results indicate that eugenol is a suitable agent for enhancing the efficacy of poor azoles against M. pachydermatis.

The chicken chorioallantoic membrane (CAM) model is an embryonic blood capillary system considered a suitable “in vivo” model for studying the irritation effect of plant extracts on the vascular system, including impacts on hemostasis, hyperemia, hemorrhage, and coagulation. The main aim of the present work was to investigate the irritation effects of different concentrated alcohol extracts of Ginkgo biloba L. (GBE) leaves on the blood vessels of the CAM model during early embryogenesis, evaluated using the Luepke scoring system. The antioxidant properties of GBE were assessed using DPPH radical scavenging and the FRAP method, alongside HPLC-DAD analysis to confirm the presence of major therapeutically relevant metabolites, revealing a strong therapeutic potential of the extract. On embryonic day 9, different concentrations of GBE as well as the controls (saline solution and 30% ethanol) were applied to the CAM surface. Vascular changes were observed immediately after application, with vasoconstriction leading to the temporary “disappearance” of blood vessels. At 30 s post-application, all GBE concentrations and ethanol induced hyperemia and mild hemorrhage, which gradually diminished over time. No changes were observed with saline application. The extent of morphometric changes in the vessels was also influenced by the concentration of GBE used. Concentrations of 20% and 30% GBE induced vasoconstriction. Lower concentrations of GBE induced vasodilation, with maximum values recorded after 240 s for 1% and 15% GBE. The results of this study may help to better characterize the vascular effects of natural Ginkgo biloba under in vivo conditions and promote greater interest in the use of alternative animal models in pharmacological and biomedical research.

Natural Ophiocordyceps sinensis is a highly valued medicinal fungus known for its antitumor, immunomodulatory, and antiviral properties. Due to extensive overharvesting in Asia, cultivated alternatives have become increasingly important. This study aimed to evaluate the biological activity and chemical composition of extracts obtained from cultivated Ophiocordyceps sinensis grown using different rice substrates. Methanolic extracts were prepared from solid-state cultivated Ophiocordyceps sinensis grown on Oryza sativa var. indica and Oryza sativa var. japonica. Antioxidant activity was determined using the DPPH assay, while proteolytic activity was evaluated with the azocasein substrate. Chemical characterization of major compounds was performed using ¹D and ²D NMR spectroscopy, together with IR spectroscopy. UV/Vis spectrophotometry was employed to confirm the formation of silver nanoparticles in AgNO₃ solution. Antimicrobial activity was tested against bacterial strains, including Escherichia coli and Staphylococcus aureus. All prepared methanolic extracts exhibited measurable antioxidant and proteolytic activities. The dominant identified compounds were Z-oleic acid, linoleic acid, and D-mannitol. Selected extracts successfully induced the formation of silver nanoparticles. The highest antimicrobial activity against Escherichia coli was observed for sample 1OS, reaching a mean % RIZD value of 129.32 ± 0.58%.