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Additional file 1: Table S1. Intralaboratory reproducibility of HPV Selfy according to Meijer’s guidelines. HPV Selfy assay was performed twice on a subpopulation of 521 samples of which 157 samples positive to HC2. Overall concordance observed was 94.6% (kappa value of 0.87). Table S2. Interlaboratory reproducibility of HPV Selfy according to Meijer’s guidelines. HPV Selfy assay was performed in another laboratory (MediChrom) on a subpopulation of 500 samples of which 150 samples positive to HC2. Overall concordance observed was 93.6% (kappa value of 0.85). Table S3. Intra-laboratory reproducibility of genotype findings of HPV Selfy. Data are presented as number of each genotype detected in each run (i.e. run1 and/or run 2), and numbers do not count up to the total number of HR-HPV positive samples due to multiple infections.

I materiali ibridi bioispirati costituiti da titanio e melanina sono molto promettenti per lo sviluppo di sistemi multifunzionali impiegati in una serie considerevole di applicazioni. Il biossido di titanio (TiO2) è un prodotto economico, ecologico e fotostabile con un forte potenziale antimicrobico. Le eumelanine sono pigmenti idrofobici con carica negativa ad elevato peso molecolare, altamente immunogeniche, esse esercitano un'azione protettiva contro microrganismi e radiazioni UV. Questo studio dimostra la capacità delle interazioni degli ioni titanio con due precursori della melanina: l'acido 5,6 diidrossiindolo-2-carbossilico (DHICA) e la diidrossifenilalanina (DOPA) nel dirigere la via melanogenica. Saggi di attività antimicrobica diretti contro il ceppo Gram-negativo di Escherichia coli DH5α hanno consentito di valutare il contributo di ciascun componente, nonché del complesso di trasferimento di carica TiO2DHICA e TiO2DOPA alle prestazioni biologiche complessive. Infine, i risultati della caratterizzazione antimicrobica sono stati combinati con quelli spettroscopici per dimostrare se e quanto il trattamento ossidativo inducesse una modificazione strutturale nella melanina, migliorando così l'efficacia antimicrobica complessiva, e ponendo le basi per la determinazione del meccanismo d'azione di questi nuovi materiali ibridi. Il polietilene (PE) è un polimero sintetico molto idrofobo, ad alto peso molecolare e difficilmente biodegradabile. È molto importante trovare microrganismi capaci di aumentare l'idrofilia del PE e/o ridurre la lunghezza della sua catena polimerica. In questo studio, il fungo Cladosporium halotolerans, è stato isolato dal sistema gastrico delle larve di Galleria mellonella. Il fungo si è dimostrato capace di degradare le microparticelle di PE, e se coltivato in presenza di questo polimero, secerne nel surnatante enzimi degradativi come la laccasi, combinando un'azione meccanica ed enzimatica che mira a modificare la struttura chimica del polietilene in un periodo di 15 giorni.

Polyethylene (PE) is high molecular weight synthetic polymer, very hydrofobic and hardly biodegradable. To increase polyethylene bio-degradability it is very important to find microorganisms that improve the PE hydrophilic level and/or reduce the length of its polymeric chain by oxidation. In this study, we isolated Cladosporium halotolerans, a fungal species, from the gastric system of Galleria mellonella larvae. Here, we show that C. halotolerans grows in the presence of PE polymer, it is able to interact with plastic material through its hyphae and secretes enzymes involved in PE degradation.

HPV Selfy clinical sensitivity and specificity resulted non-inferior to those of HC2. By analysis of a total of 889 cervical liquid-based cytology samples from a screening population, of which 98 were from women with CIN2+, HPV Selfy showed relative sensitivity and specificity for CIN2+ of 0.98 and 1.00 respectively (non-inferiority score test: P = 0.01747 and P = 0.00414, respectively); the test reached adequate intra- and inter-laboratory reproducibility. Moreover, we demonstrated that the performance of HPV Selfy on self-collected vaginal samples was non-inferior to the performance obtained on clinician-collected cervical specimen (0.97 relative sensitivity and 0.97 relative specificity). Finally, through HPV Selfy genotyping, we were able to describe HPV types prevalence in the study population. Clinical validation of HPV Selfy on self‑collected vaginal samples (VALHUDES) Next, we aimed at evaluating HPV Selfy performance on self-collected samples, as indicated by VALHUDES protocol. Hence, we needed to assess whether HPV testing on vaginal self-samples was as accurate as HPV testing on a cervical sample taken by a clinician. To do so, we identified 119 CIN2+ cases (age 25–65 years) and 791 ≤ CIN1 cases, for which we had available paired cervical specimen and self-collected vaginal samples. HPV Selfy testing in self-collected samples was found similarly sensitive (93/96; relative sensitivity 0.97; 95% CI 0.90–1.04) and specific (722/745; relative specificity 0.97; 95% CI 0.95–0.99) to detect CIN2+ in the total study population (Table 4), in comparison with HPV Selfy performed on paired ThinPrep. Thus, HPV Selfy assay fulfills VALHUDES requirements for use of HR-HPV DNA tests on self-collected samples according to non-inferiority analysis (relative sensitivity > 0.90 with T = 1.70, p = 0.044; relative specificity > 0.95 with T = 1.87, p =0.031). Secondary objectives of VALHUDES protocol include the assessment of the absolute accuracy of HR-HPV DNA test applied according to the sampling device and the proportion of adequate samples as determined by amplification of an internal control (a ubiquitous human gene). HPV Selfy assay provides a human beta-globin internal control, used to evaluate sample quality. In the whole study cohort, mean Ct value for the human beta-globin internal control for the HPV Selfy test on self-collected samples, obtained with the direct analysis on self-collected samples, was 26.1 Ct (median value 25.9 Ct, maximum 30.7 Ct, minimum 16.5 Ct). In the subgroup of women with biopsy-diagnosis of cervical lesions CIN2+, the same analysis resulted in 26.7 Ct (median value 26.5 Ct, maximum 30.7 Ct, minimum 24.2 Ct). This means that all women were able to self-collect a similar amount of specimen, confirming self-collected samples’ quality adequacy and so the easiness of the self-sampling procedure using the FLOQSwabs® (Copan, Brescia, Italy).

Daucus is a genus of economically important plants belonging to Apiaceae family spread in temperate regions. Species of this genus are used as food and several biological properties have reported. The chemical composition of the essential oils from different organs (roots, stems and flowers) of Daucus carota subsp. maximus, a species not previously investigated, was analyzed by GC-MS. Our results showed the presence of β-phellandrene as the most abundant component of stems and flowers and of γ-terpinene as a major compound of the oil from the roots. Flower essential oil caused a greater increase in the activity of superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPX) in polymorphonuclear leukocytes (PMN) cells compared to stem and root essential oils. The antimicrobial activity of the flower and stem oil were more effective, compared to root oil, against Escherichia coli and Staphylococcus aureus.

Foeniculum vulgare (Apiaceae) is an aromatic fennel with important practices in medicinal and traditional fields, used in the treatment of digestive complications, and gastrointestinal and respiratory disorders. Its leaves and stems, tender and fresh, are used in the production of pasta dressing and main courses, while its seeds, with a strong smell of anise, are excellent flavoring for baked goods, meat dishes, fish, and alcoholic beverages. The aim of this work is concerning the extraction of essential oil (EO) from the leaves of F. vulgare subsp. vulgare var. vulgare, investigating antimicrobial, antibiofilm, and antioxidant efficacy. In particular, GC-MS analysis showed how the chemical composition of EO was influenced by the massive presence of monoterpene hydrocarbons (α-pinene 33.75%) and phenylpropanoids (estragole 25.06%). F. vulgare subsp. vulgare var. vulgare EO shows excellent antimicrobial activity against both Gram-positive and Gram-negative strains. This EO can inhibit biofilm formation at very low concentrations and has a good ability to scavenge oxygen radicals in vitro. F. vulgare subsp. vulgare var. vulgare EO also has an increased activity of superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx) enzymes and decreased ROS levels in zymosan opsonized PMNs (OZ).

In this work, the essential oil of the aerial parts of a not previously investigated accession of Prangos trifida (Mill.) Herrnst. & Heyn (Apiaceae), collected in Northern Italy, was analysed by GC/MS. Twenty-five compounds were identified, accounting for 91.30% of the total oil, which is characterised by a high content of monoterpene hydrocarbons (71.26%). The main constituents of the essential oil were cis-beta-ocimene (18.12%), alpha-phellandrene (12.14%), sylvestrene (11.32%), p-mentha-1,3,8-triene (9.56%) and alpha-pinene (8.85%). The essential oil of P. trifida showed a good antimicrobial activity directed above all against Gram-positive bacteria, and had an interesting antioxidant activity, thus making it an excellent candidate for future cosmetic and pharmaceutical applications.

Prangos ferulacea (L.) Lindl, which belongs to the Apiaceae family, is a species that mainly grows in the eastern Mediterranean region and in western Asia. It has been largely used in traditional medicine in several countries and it has been shown to possess several interesting biological properties. With the aim to provide new insights into the phytochemistry and pharmacology of this species, the essential oils of flowers and leaves from a local accession that grows in Sicily (Italy) and has not yet been previously studied were investigated. The chemical composition of both oils, obtained by hydrodistillation from the leaves and flowers, was evaluated by GC-MS. This analysis allowed us to identify a new chemotype, characterized by a large amount of (Z)-β-ocimene. Furthermore, these essential oils have been tested for their possible antimicrobial and antioxidant activity. P. ferulacea essential oils exhibit moderate antimicrobial activity; in particular, the flower essential oil is harmful at low and wide spectrum concentrations. They also exhibit good antioxidant activity in vitro and in particular, it has been shown that the essential oils of the flowers and leaves of P. ferulacea caused a decrease in ROS and an increase in the activity of superoxide dismutase (SOD), catalase (CAT) and glutathione S-transferase (GST) in OZ-stimulated PMNs. Therefore, these essential oils could be considered as promising candidates for pharmaceutical and nutraceutical preparations.

The Stachys L. genus has been used in traditional medicine to treat skin inflammations, stomach disorders, and stress. The aim of this study was to investigate the chemical profile and biological activity of the methanolic extract of Stachys spreitzenhoferi Heldr. (Lamiaceae) aerial parts, collected on the island of Kythira, South Greece. The analysis by liquid chromatography coupled with electrospray ionization and high-resolution mass spectrometry [LC-(-)ESI/HRMSn] of the methanol extract revealed the occurrence of thirty-six compounds - flavonoids, phenylethanoid glycosides, iridoids, quinic acid derivatives, aliphatic alcohol glycosides, and oligosaccharides - highlighting the substantial presence, as main peaks, of the iridoid melittoside (2) along with flavonoid compounds such as 4'-O-methylisoscutellarein mono-acetyl-diglycoside/chrysoeriol mono-acetyl-diglycoside (24), trimethoxy- (35) and tetramethoxyflavones (36). This extract was tested for its antimicrobial properties against Gram-positive and negative pathogenic strains. The extract was not active against Gram-negative bacteria tested, but it possessed a good dose-dependent antimicrobial activity towards S. aureus (MIC: 1.0mg/mL) and L. monocytogenes (MIC: 1.0mg/mL) Gram-(+) strains. Furthermore, this extract has been tested for its possible antioxidant activity in vitro. In particular, it has been shown that these molecules cause a decrease in DPPH, ABTS, and H2O2 radicals. The extract of S. spreitzenhoferi exhibited anti-DPPH activity (IC50: 0.17mg/mL), anti-H2O2 activity (IC50: 0.125mg/mL), and promising antiradical effect with an IC50 value of 0.18mg/mL for anti-ABTS activity. S. spreitzenhoferi extract caused a decrease in ROS (at the concentration of 200μg/mL) and an increase in the activity of the antioxidant enzymes SOD, CAT, and GPX in OZ-stimulated PMNs. Furthermore, it exhibited antiproliferative activity against acute myeloid leukemia (U937cell), causing 50% of cell death at the 0.75mg/mL.

Background According to international guidelines, Human Papillomavirus (HPV) DNA tests represent a valid alternative to Pap Test for primary cervical cancer screening, provided that they guarantee balanced clinical sensitivity and specificity for cervical intraepithelial neoplasia grade 2 or more (CIN2+) lesions. The study aimed to assess whether HPV Selfy (Ulisse BioMed – Trieste, Italy), a full-genotyping HPV DNA test that detects and differentiates 14 high-risk HPV (HR-HPV) types, meets the criteria for primary cervical cancer screening described in the international guidelines, on clinician-collected as well as on self-collected samples. Methods For each participant woman, consecutively referring to Azienda Sanitaria Universitaria Giuliano Isontina (Trieste, Italy) and CRO—National Cancer Institute (Aviano, Italy) for the cervical cancer screening program, the following samples were tested: (a) a clinician-collected cervical specimen, analyzed with the reference test (Hybrid Capture®2 test, HC2) and HPV Selfy; and (b) a self-collected vaginal sample, analyzed with HPV Selfy. Enrolled women were also asked to fulfill a questionnaire about self-sampling acceptability. As required by guidelines, a non-inferiority test was conducted to compare the clinical performance of the test under evaluation with its reference test. Results HPV Selfy clinical sensitivity and specificity resulted non-inferior to those of HC2. By analysis of a total of 889 cervical liquid-based cytology samples from a screening population, of which 98 were from women with CIN2+, HPV Selfy showed relative sensitivity and specificity for CIN2+ of 0.98 and 1.00 respectively (non-inferiority score test: P = 0.01747 and P = 0.00414, respectively); the test reached adequate intra- and inter-laboratory reproducibility. Moreover, we demonstrated that the performance of HPV Selfy on self-collected vaginal samples was non-inferior to the performance obtained on clinician-collected cervical specimen (0.92 relative sensitivity and 0.97 relative specificity). Finally, through HPV Selfy genotyping, we were able to describe HPV types prevalence in the study population. Conclusions HPV Selfy fulfills all the requirements of the international Meijer’s guidelines and has been clinically validated for primary cervical cancer screening purposes. Moreover, HPV Selfy has also been validated for self-sampling according to VALHUDES guidelines. Therefore, at date, HPV Selfy is the only full-genotyping test validated both for screening purposes and for self-sampling. Trial registration ASUGI Trieste n. 16008/2018; CRO Aviano n.17149/2018

Acca sellowiana is an evergreen plant that produces edible fruit whit high nutritional properties, and also contains clinically relevant bioactive compounds, including polyphenols and essential oils. Numerous biological activities were demonstrated for A. sellowiana fruits: antifungal and antitumoral and anti-oxidant. It was also showed a strong antibacterial activity against Gram-positive and Gram-negative bacteria. Fruit are generally considered recalcitrant plant tissues for the difficulty to obtain high quality protein due to a low protein content and the presence of interfering substances. For this reason, the objective of the present work was obtain high quantity of protein extract, to determine the location in the fruit of the molecules responsible for the antibacterial activity, separate them according to molecular weight, test their thermo resistance, study the timing of action, isolate the protein fraction having activity and hypothesise a mechanism of action directed against bacterial membranes.

The peculiar aspect that emerges from the study of Orchidaceae is the presence of various molecules, which are particularly interesting for pharmaceutical chemistry due to their wide range of biological resources. The aim of our study was to investigate the properties of two dihydrophenanthrenes, isolated, for the first time, from Himantoglossum robertianum (Loisel.) P. Delforge (Orchidaceae) bulbs and roots. Chemical and spectroscopic study of the bulbs and roots of Himantoglossumrobertianum (Loisel.) P. Delforge resulted in the isolation of two known dihydrophenanthrenes—loroglossol and hircinol—never isolated from this plant species. The structures were evaluated based on 1H-NMR, 13C-NMR, and two-dimensional spectra, and by comparison with the literature. These two molecules have been tested for their possible antioxidant, antimicrobial, antiproliferative, and proapoptotic activities. In particular, it has been shown that these molecules cause an increase in the activity of superoxide dismutase (SOD), catalase (CAT), and glutathione S-transferase (GST) in polymorphonuclear leukocytes (PMN); show antimicrobial activity against Escherichia coli and Staphylococcus aureus, and have anti-proliferative effects on gastric cancer cell lines, inducing apoptosis effects. Therefore, these two molecules could be considered promising candidates for pharmaceutical and nutraceutical preparations.