Dario Antonini

The complex genus Achillea L. comprises more than 140 species distributed widely throughout the Northern Hemisphere. Several species are widely used in traditional medicine for their therapeutic properties, yet few studies have correlated their biological properties with the plant’s phytochemical composition. Among these, Achillea setacea Waldst. & Kit. is a perennial species traditionally used to treat digestive and inflammatory disorders. In this study, the essential oil of A. setacea, collected wild in North Macedonia, was analyzed spectrometrically and spectroscopically by GC-MS and NMR, respectively. A total of nineteen compounds were identified, with camphor (31.3%), 4-terpineol (11.3%), and eucalyptol (10.6%) being the main constituents. Furthermore, the biological activities of pure oil were evaluated, showing notable antioxidant properties, as well as antimicrobial effects against a panel of clinically relevant microorganisms, including Gram-positive and Gram-negative bacteria. Furthermore, its impact on human intestinal epithelial (Caco-2) cells was assessed, highlighting its potential relevance for gastrointestinal applications, in agreement with the traditional use of Achillea species for digestive disorders.

The tumor suppressor ARF (p14 in human, p19 in mouse), has traditionally been characterized by its pivotal role in tumor surveillance. However, its involvement in an expanding range of cellular processes reveals that its functions are broader and more complex than initially appreciated. Here, we uncover a previously unrecognized role of p19ARF in endodermal differentiation, specifically in pancreatic lineage specification using an in vitro differentiation model of mouse embryonic stem cells (mESCs). Using CRISPR/Cas9-mediated mutagenesis, we show that mESCs with mutations in p19Arf are unable to efficiently differentiate towards pancreatic endoderm. Transcriptomic profiling reveals substantial alterations in gene networks associated not only with lineage commitment but also with cytoskeletal organization and cell morphology. These changes correlate with a disruption in stem cell architecture and suggest the persistence of pluripotency feature when only one copy of functional p19Arf is present. Taken together, our findings highlight a role for p19Arf in modulating endodermal differentiation while maintaining the essential cellular properties of stem cells, thus expanding its relevance beyond tumor suppression.

Aberrant transcriptional regulation is a defining feature of squamous cell carcinoma (SCC), yet how lineage transcription factors coordinate shared and factor-specific oncogenic programs remains poorly understood. Although TP63 (p63) is frequently amplified in SCC, the contribution of its paralog TP73 (p73) has remained unclear. Here we show that p73, together with p63, is upregulated in skin SCC and is required for tumorigenesis. Mechanistically, p63 and p73 form heteromeric complexes and co-occupy distal enhancer elements, establishing a shared chromatin regulatory framework. Integration of chromatin and transcriptomic profiling reveals that this common enhancer landscape supports both convergent and divergent transcriptional outputs. Both factors cooperatively sustain core proliferation but also exert regulatory biases, with p63 preferentially reinforcing epithelial lineage circuits and p73 contributing to DNA replication and stress-associated pathways. Among shared downstream targets, p63/p73 co-regulation of multiple epidermal growth factor receptor (EGFR) ligands establishes a feed-forward signaling module that amplifies mitogenic signaling. Amphiregulin emerges as a dominant functional mediator, and its depletion phenocopies key aspects of p63/p73 loss, including impaired proliferation and tumor formation. Together, these findings support a model in which shared enhancer occupancy by p63 and p73 drives cooperative and factor-specific transcriptional programs, linking chromatin regulation to signaling and tumor maintenance.

The increasing problem of antibiotic resistance is a critical global health issue, necessitating the development of alternative therapeutic strategies to manage infections effectively. Among the promising solutions are human antimicrobial peptides (AMPs), naturally occurring molecules known for their broad spectrum of antimicrobial activity. Background/Objectives: This study investigates the potential of some AMPs, selected through a bioinformatic approach, as alternatives to conventional antibiotics, particularly focusing on their efficacy against species within the Bacteroidota phylum. These species, including pathogens such as Porphyromonas gingivalis, Capnocytophaga ochracea, and Capnocytophaga canimorsus, are well known for their roles in various human infections and related diseases. Non-pathogenic environmental species, such as Flavobacterium johnsoniae, are also included in this group, frequently used as a model organism. Methods: By analyzing the antimicrobial efficacy, mechanisms of action, and potential therapeutic applications of human AMPs, this research underscores their significance in addressing the challenge of antibiotic resistance. Results: This study identified three peptides, KTL24, LIR23, and MFP22, as particularly interesting. These peptides are derived from specific human proteins, namely SPI1, NAPSA and SCUB1. Conclusions: Their notable antimicrobial potential suggests that AMPs could serve either as a complementary treatment alongside traditional antibiotics or as a standalone therapy, mitigating the ongoing spread of antibiotic resistance and offering an alternative in global health strategies.

Daucus is a large genus of the Apiaceae family, comprising around forty-five accepted species, that has a worldwide distribution. Species of this genus have been reported to have several traditional medicinal uses, and some of them are also largely used as food and spices. Daucus nebrodensis Strobl. is an endemic species of Sicily growing in the montane environments of the Madonie and the Nebrodi Mountains. In this work, the essential oil of D. nebrodensis (DnEO), collected wild near Messina (Italy), was chemically and biologically investigated. The hydrodistilled essential oil (yield 0.15%), obtained from fresh aerial parts, was evaluated by GC-MS, and It was particularly rich in monoterpene hydrocarbons, with sabinene (33.6%), α-pinene (17.2%), γ-terpinene (9.8%), and α-terpinene (7.6%) as the main metabolites. DnEO, and its main constituents, have been tested to evaluate their biological properties. Given the current problem of antibiotic resistance, it is of great interest to identify alternative molecules that could counteract the its progression. Therefore, DnEO was tested against Gram-negative species, such as E. coli DH5α and P. aeruginosa PAOI, and Gram-positive species, such as S. aureus ATCC6538P, B. subtilis AZ54, and M. smegmatis MC²155, showing notable antibacterial activity. The MIC for Bacillus subtilis, the most sensitive strain, was 18 mg/mL, while the MIC for Pseudomonas aeruginosa, the least sensitive strain, was 30 mg/mL. Moreover, interesting antibiofilm activity was observed against Mycobacterium smegmatis with a 55% inhibition. Its ability to form biofilms contributes to its persistence and resistance in clinical settings. These findings highlight the potential of D. nebrodensis EO as a source of bioactive compounds with promising antimicrobial and antibiofilm properties.

Mutations in the TP63 gene cause several syndromic disorders, including ankyloblepharon–ectodermal defects–cleft lip/palate (AEC) syndrome, characterized by severe skin erosions, cleft palate, and ectodermal dysplasia. These mutations often affect the carboxy-terminal sterile-α-motif (SAM) domain of the p63 protein, leading to domain misfolding, protein aggregation, and impaired transcriptional activity. To dissect the molecular mechanisms underlying AEC pathogenesis, we investigated primary keratinocytes derived from p63L514F mutant mice, which carry a SAM domain mutation associated with AEC syndrome. p63L514F keratinocytes exhibited significantly reduced proliferation compared to wild-type controls, as indicated by decreased 5-ethynyl-2′-deoxyuridine (EdU) incorporation, decreased Cyclin D1 and Cyclin D2 expression, and an increase in the cell-cycle inhibitors p21 and p27. Furthermore, p63L514F keratinocytes showed increased cell death, elevated reactive oxygen species (ROS) levels, and a decreased reduced (GSH) and oxidized (GSSG) glutathione (GSH/GSSG) ratio, indicating oxidative stress. This stress response was accompanied by a marked reduction in Solute Carrier Family 7 Member 11 (Slc7a11), a critical regulator of antioxidant defense. We further identified Slc7a11 as a likely direct transcriptional target of p63: p63 depletion reduced Slc7a11 expression, and chromatin immunoprecipitation uncovered an evolutionary conserved p63-binding enhancer upstream of the Slc7a11 promoter. Together, our findings demonstrate that p63 mutations causative of AEC syndrome impair keratinocyte proliferation, promote cell death via oxidative stress, and compromised antioxidant defenses, revealing a dual role for p63 in sustaining skin homeostasis.

The genus Lavandula L., belonging to the Lamiaceae family, contains about forty species with a distribution that mainly extends in the Mediterranean basin from the North Atlantic to the Middle East. Due to their excellent biological properties, the aerial parts and flowers of Lavandula peduncolata ssp. have been utilized in traditional medicine in Morocco and western Europe. This paper investigated the chemical composition and antibacterial activities as well as the antibiofilm and antioxidant activities of the essential oil (EO) obtained from the pre-flowering aerial parts of Lavandula pedunculata subsp. atlantica collected in Morocco. The chemical composition of the EO, obtained by classic hydrodistillation, showed by GC-MS analysis the presence of a large amount of oxygenated monoterpene compounds. The main constituents of the EO were camphor (27.8%), camphene (10.9%), fenchone (10.6%), and eucalyptol (8.5%). The EO has been evaluated for its antimicrobial, antibiofilm, antioxidant and anti-inflammatory properties, showing promising activity against both Gram-positive and Gram-negative strains. These findings highlight the potential of Lavandula EO in combating infections caused by Streptococcus mutans and Streptococcus oralis (oral diseases), Staphylococcus aureus (skin infections), Escherichia coli and Shigella sonnei (gastrointestinal and urinary infections). In addition, although the EO showed no evident effects on cell viability in eukaryotic epithelial cells, it exhibited promising effects on anti-inflammatory properties.

Pulicaria species are used as herbal medicine and in the preparation of decoctions in several Asian and African regions. Among them, the plant Pulicaria burchardii is known for its medicinal properties, but comprehensive studies on its biological activity are still limited. This study examined the properties of the essential oil (EO) extracted by P. burchiardii and collected in Morocco during the flowering period. The focus was on its antimicrobial, anti-germinative, antioxidant, and antiviral activities, with the aim of evaluating its potential use in food preservation and beyond. The EO was subjected to various analyses to determine its chemical composition and biological efficacy. Specifically, GCMS and NMR analyses revealed that the EO is rich in oxygenated sesquiterpenes (72.59%), with 1-oxo-bisabolone being the predominant component (65.09%). The antimicrobial activity was tested against various Gram-positive and Gram-negative bacteria, demonstrating a significant inhibition of bacterial growth, particularly against Bacillus subtilis (MIC value of 0.6 mg/mL). The anti-germinative property was evaluated on spores of B. subtilis and other bacilli, such as Bacillus cereus, revealing a notable ability to prevent germination. For antiviral activity, the EO was tested against several pathogenic viruses including SARS-CoV-2 and HSV-1, showing an effective broad-spectrum reduction in viral replication in vitro. This study demonstrated that P. burchardii essential oil had excellent antibacterial and antiviral capabilities. The future challenge will focus mainly on the principal compound, 1-oxo-bisabolone, to demonstrate its real effectiveness as an antibacterial and/or antiviral.

LAMA2, coding for the laminin-α2 chain, is a crucial ECM component, particularly abundant in skeletal muscle. Mutations inLAMA2trigger the often-lethalLAMA2-congenital muscular dystrophy (LAMA2-CMD). Various phenotypes have been linked to LAMA2-CMD; nevertheless, the precise mechanisms that malfunction during disease onset in utero remain unknown. We generatedLama2-deficient C2C12 cells and found thatLama2-deficient myoblasts display proliferation, differentiation, and fusion defects, DNA damage, oxidative stress, and mitochondrial dysfunction. Moreover, fetal myoblasts isolated from thedy^Wmouse model of LAMA2-CMD display impaired differentiation and fusion in vitro. We also showed that disease onset during fetal development is characterized by a significant down-regulation of gene expression in muscle fibers, causing pronounced effects on cytoskeletal organization, muscle differentiation, and altered DNA repair and oxidative stress responses. Together, our findings provide unique insights into the critical importance of the laminin-α2 chain for muscle differentiation and muscle cell homeostasis.