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The current state of research on the anti‑SARS‑CoV‑2 potential of artemisinin‑related compounds has identified arteannuin B as a potent inhibitor of the nCoV‑2019BetaCov/Wuhan/WiV04/2019 and BetaCov/Italy/CDG1/2020 strains of the virus. The aim of this work was to fractionate the targeted sesquiterpenoid compounds, arteannuin B and artemisinin, from the complex matrix of the crude ethanolic leaf extract of Artemisia annua L. using high‑speed countercurrent chromatography (HSCCC) and to test the simplified or purified fractions against the genomically characterized Alpha SARS‑CoV‑2 variant in vitro. This is the first detailed in vitro anti‑SARS‑CoV‑2 study using an analytically characterized supercritical fluid extract of A. annua L. The preparative HSCCC method enabled the isolation of purified arteannuin B in a single chromatographic step, which was confirmed by LC‑ESI‑QTOF‑MS/MS. The MS data confirmed the selectivity of the HSCCC method for the targeted fractionation of artemisinin from the complex matrix, as it was successfully separated from the EtOH crude extract without co‑elution with arteannuin B. Antiviral activity determined by quantitative real‑time PCR (qRT‑PCR) yielded half‑maximal effective concentrations (EC₅₀) of 93.7 µg/mL (SC‑CO₂ extract), 173.5 µg/mL (EtOH extract), 187.3 µg/mL (artemisinin knockout fraction), 38.1 µg/mL (arteannuin B fraction), and >100 µg/mL (artemisinin). The arteannuin B fraction was highly active at 50 µg/mL (p < 0.0001) and 100 µg/mL (p < 0.0001), and inhibited the amplification of the SARS‑CoV‑2 N and RdRp genes by 84% and 100%, respectively. An important contribution of this study is the demonstration of the antiviral activity of arteannuin B against the Alpha variant of SARS‑CoV‑2, which is known to have increased infectivity and transmissibility.

Emina Koric, Violeta Milutinović, Alma Hajrudinović-Bogunić, F. Bogunić, Tatjana Kundaković-Vasović, Irma Gusic, Jelena Radović Selgrad, K. Durić et al.

The genetic, morphological and taxonomic diversity of the genus Sorbus is due to homoploid and polyploid hybridisation, autopolyploidy and apomixis, which also influence the production and diversity of secondary metabolites, especially flavonoids. The aim of this study was to investigate the relationships and variations of flavonoids in terms of hybrid origin and ploidy level between the parental species and their hybrid derivatives. The sampling design included leaf material of the following Sorbus accessions from ten natural localities: parental taxa (di-, tri- and tetraploids of S. aria; diploid S. torminalis and S. aucuparia) and their di-, tri- and tetraploid hybrid derivatives from crosses of S. aria × S. torminalis (subg. Tormaria) as well as the tetraploid S. austriaca and S. bosniaca, which originate from crosses of S. aria × S. aucuparia (subg. Soraria). We analysed the flavonoid profiles from the leaf fractions by LC-MS. A total of 23 flavonoids were identified, including apigenin and luteolin derivatives, which distinguish the hybrid groups from each other. This profiling highlights the distinctiveness of the Tormaria and Soraria accessions and emphasises the potential of the subg. Tormaria for further research on bioactive compounds in biological studies.

Introduction: Rheumatoid arthritis (RA) is a chronic inflammatory disease characterized by joint inflammation and destruction, leading to significant pain and disability. Adenosine deaminase (ADA) is identified as a biomarker for RA’s inflammatory process. This study aims to investigate the potential of flavonoids and phenolic acids to inhibit ADA activity (in silico) and evaluate their anti-inflammatory effects in a RA model (in vivo). Methods: The molecular docking study was conducted using YASARA Structure 19.12.14. software following the Auto Dock 4.2 protocol. A rat model with pristane-induced arthritis was used to test the anti-inflammatory effect of selected polyphenols. The consistency of the development of the rat model was evaluated through the following indicators artistic score, paw volume, and body weight. Quercetin was administered intragastrically at doses of 150 and 400 mg/kg over 15 days. The C-reactive protein (CRP) level in serum was measured with an automatic biochemical analyzer. Statistical analyses were performed using SPSS 29.0.2.0. Results: Molecular docking simulations showed flavonoids inhibited ADA activity with inhibition constants ranging from 0.012 mM to 0.190 mM. In the in vivo RA model, quercetin significantly reduced joint inflammation and serum CRP levels at a higher dose of 400 mg/kg. Conclusion: Quercetin shows promise as an anti-inflammatory agent for RA by targeting ADA, suggesting that flavonoid-rich plant extracts could enhance RA treatment.

Abstract The study aimed to analyse the potential of Lamiaceae essential oils, extracts, and hydrolats against bacterial biofilms. Bacterial cells Pseudomonas aeruginosa, Staphylococcus aureus, and Bacillus cereus were exposed to Thymus vulgaris L. (thyme), Salvia officinalis L. (sage), Mentha × piperita L. (mint) essential oils, extracts, and hydrolats. The result of the minimal inhibitory concentration assessment shows the highest antibacterial potential for essential oils, followed by extracts and hydrolats respectively. The anti-biofouling capacity revealed that thyme essential oil has the highest potential for biofilm prevention for all tested bacteria, reducing up to 91% of biofilm, followed by mint (88%) and sage (87%) essential oil. While the thyme extract (84%), sage extract (83%) and hydrolat (77%) we less effective. The chemical composition of thyme essential oil showed a high percentage of monoterpene hydrocarbons and oxygenated monoterpenes, among which p-cymene and thymol were the most predominant. The bacterial cell membrane integrity assessment shows a significant increase in dead cells by increasing the concentration of thyme essential oil. The findings of our research indicate that the choice of herbal preparation significantly affects the active components, thereby influencing both antibacterial and anti-biofouling capabilities. Lamiaceae essential oils show great potential for biofilm management and represent a good candidate for antibacterial application in pharmacy, medicine, and industry.

Triterpenes are very important secondary metabolites with wide structural diversity and significant role in pharmacy and medicine.In the present research, a comparative study of pharamacological activities of the triterpene fractions obtained from several plant species belonging to Lamiaceae family, was carried out. In-vitro anti-proliferative activity was performed using a standardproliferation assay based on tetrazolium salts. In vitro anti-inflammatory activity of triterpene fractions was determined by an assay of inhibition of albumin denaturation. In general, the triterpene fractions obtained from plant species belonging to Lamiaceae family showed a strong anti-proliferative activity and anti-inflammatory activity.The triterpene fraction of Rosmarini folium showed the strongest anti-proliferative activity (GI50range from 4 to 37 μg/ml) and the strongest anti-inflammatory activity in the range from 57.27% to 80.69%. This comparative study provides scientific evidence to support the traditional use of Lamiacae plant species for medical purposes as anti-inflammatory and anti-proliferative medicines.

Among natural products, essential oils from aromatic plants have been reported to possess potent anticancer properties. In this work, we aimed to perform the cytotoxic concentration range screening and antiproliferative activity screening of chemically characterized Thymus vulgaris L. essential oil. In vivo bioassay was conducted using the brine shrimp lethality test (BSLT). In vitro evaluation of antiproliferative activity was carried out on three human tumor cell lines: breast adenocarcinoma MCF-7, lung carcinoma H460 and acute lymphoblastic leukemia MOLT-4 using MTT assay. Essential oil components thymol (36.7%), p-cymene (30.0%), γ-terpinene (9.0%) and carvacrol (3.6%) were identified by gas chromatography/mass spectrometry. Analyzed essential oil should be considered as toxic/highly toxic with LC50 60.38 µg/mL in BSLT and moderate/weakly cytotoxic with IC50 range 52.65–228.78 µg/mL in vitro, according to evaluated cytotoxic criteria. Essential oil induced a dose-dependent inhibition of cell proliferation in all tested tumor cell lines and showed different sensitivity. Dose dependent toxicity observed in bioassay as well as the in vitro assay confirmed that brine shrimp lethality test is an adequate method for preliminary toxicity testing of Thymus vulgaris L. essential oil in tumor cell lines.

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