Functionalized PLGA-Based Nanoparticles with Anti-HSV-2 Human Monoclonal Antibody: A Proof of Concept for Early Diagnosis and Targeted Therapy

Background: Functionalized nanoparticles (NPs) represent a cutting edge in innovative clinical approaches, allowing for the delivery of selected compounds with higher specificity in a wider time frame. They also hold promise for novel theranostic applications that integrate both diagnostic and therapeutic functions. Pathogens are continuously evolving to try to escape the strategies designed to treat them. Objectives: In this work, we describe the development of a biotechnological device, Nano-Immuno-Probes (NIPs), for early detection and infections treatment. Human Herpes Simplex Virus 2 was chosen as model pathogen. Methods: NIPs consist of PLGA-PEG-Sulfone polymeric NPs conjugated to recombinant Fab antibody fragments targeting the viral glycoprotein G2. NIPs synthesis involved multiple steps and was validated through several techniques. Results: DLS analysis indicated an expected size increase with a good polydispersity index. Z-average and z-potential values were measured for PLGA-PEG-Bis-Sulfone NPs (86.6 +/- 10.9 nm; -0.7 +/- 0.3 mV) and NIPs (151 +/- 10.4 nm; -5.1 +/- 1.9 mV). SPR assays confirmed NIPs' specificity for the glycoprotein G2, with an apparent KD of 1.03 +/- 0.61 mu M. NIPs exhibited no cytotoxic effects on VERO cells at 24 and 48 h. Conclusions: This in vitro study showed that NIPs effectively target HSV-2, suggesting the potential use of these nanodevices to deliver both contrast agents as well as therapeutic compounds

VP6-SUMO Self-Assembly as Nanocarriers for Gastrointestinal Delivery

High proteolytic degradation and poor absorption through epithelial barriers are major challenges to successful oral delivery of therapeutics. Nanoparticle platforms can enhance drug stability and extend the residence time in gastrointestinal (GI) tract. However, drug delivery systems are often inactivated in acidic environment of stomach or suffer poor absorption from intestinal cells due to the mucus layer. To overcome these issues we developed a drug delivery system constituted by a protein construct made by a Rotavirus capsid protein (VP6) and the small ubiquitin-like modifier SUMO. This chimeric construct allows specificity towards intestinal cells, the Rotavirus natural target, combined by an enhanced stability given by the eukaryotic protein transporter SUMO. Furthermore SUMO can act as a molecular switch that facilitates import/export of its ligand to the nucleus, the hypersensitive subcellular site target of many cell killing therapies. In this paper we show that SUMO-VP6 constructs self-assembly into stable nanocarriers. SUMO-VP6 nanocarriers display ideal features for drug delivery: a small size and high monodispersity, a high stability in different pH conditions and a high uptake in the nuclear and cytoplasmic compartment of intestinal cells. These features make SUMO-VP6 nanocarriers a promising novel system for oral delivery of poorly soluble drugs

The swine waste resistome: Spreading and transfer of antibiotic resistance genes in Escherichia coli strains and the associated microbial communities

The overuse of antimicrobials in livestock farming has led to the development of resistant bacteria and the spread of antibiotic-resistant genes (ARGs) among animals. When manure containing these antibiotics is applied to agricultural fields, it creates a selective pressure that promotes the acquisition of ARGs by bacteria, primarily through horizontal gene transfer. Most research on ARGs focuses on their role in clinical antibiotic resistance and their transfer from environmental sources to bacteria associated with humans, such as Escherichia coli. . The study investigates the spread of antibiotic-resistant genes (ARGs) through class 1 integrons in 27 Escherichia coli strains from pig manure. It focuses on six common ARGs (ermB, erm B, cml A, flo R, qnr S, tet A, and TEM) and the class 1 integron gene, assessing their prevalence in manure samples from three pig farms. The study found correlations and anticorrelations among these genes, indicating a predisposition of the integron in spreading certain ARGs. Specifically, cmlA A and tetA A genes were positively correlated with each other and negatively with int1, suggesting they are not transferred via Int 1. Farm B had the highest int1 counts and a higher abundance of the TEM gene, but lower levels of cmlA A and tetA A genes. The results underscore the complexity of predicting ARG spread in agricultural environments and the associated health risks to humans through the food chain. The study's results offer valuable insights into the antibiotic-resistant genes (ARGs) profile in swine livestock, potentially aiding in the development of methods to trace ARGs in the environment

Potenziale utilizzo di idrolati per il controllo delle infezioni microbiche

Introduzione. Come \ue8 noto, diversi microrganismi costituiscono un diretto pericolo per la nostra salute in quanto possono infettare l'ospite umano: le malattie infettive causate da batteri e funghi sono ogni anno responsabili di morbilit\ue0 e mortalit\ue0 in un numero crescente di individui ospedalizzati e immunocompromessi. Inoltre, negli ultimi anni, la diffusione di ceppi microbici multi-resistenti ai farmaci, (ad es. Staphylococcus aureus, Pseudomonas aeruginosa e Candida spp.), ha provocato infezioni difficili o impossibili da controllare con antimicrobici convenzionali. La necessit\ue0 di nuove terapie per il trattamento delle infezioni microbiche ha portato i ricercatori a concentrarsi su possibili alternative di tipo naturale. L'idrolato (Id), noto anche come idrosol o acqua aromatica, \ue8 la frazione idrofila ottenuta durante il processo di distillazione in corrente di vapore assieme all\u2019olio essenziale (OE), capace di esercitare attivit\ue0 anti-microbiche grazie al contenuto in componenti terpeniche pari a circa l\u20191% v/v. Sebbene entrambe i prodotti della distillazione abbiano una nota azione antimicrobica, non \ue8 mai stata paragonata la loro attivit\ue0 quando estratti dalla stessa pianta. Scopo. Gli scopi di questo lavoro sono stati: (i) quello di paragonare l'attivit\ue0 antimicrobica dell\u2019OE e dell\u2019Id estratti da Monarda citriodora (MC) verso 30 ceppi fungini e batterici coinvolti in infezioni umane, (ii) valutare l\u2019efficacia dell\u2019Id di C. aurantium var. amara (CA) nei confronti di 18 ceppi fungini e batterici potenzialmente responsabili di infezioni cutanee. Materiali e metodi. L\u2019OE e l\u2019Id di MC sono stati ottenuti mediante distillazione in corrente di vapore partendo da piante fiorite coltivate presso l\u2019istituto Scarabelli-Ghini di Imola (fig.1), mentre l\u2019Id-CA era di tipo commerciale. Sono stati eseguiti test di micro-brodo diluizione secondo linee guida internazionali EUCAST per valutare l\u2019efficacia antimicrobica dell\u2019OE-MC (concentrazioni comprese tra 0,0078% e 4%), dell'Id-MC (concentrazioni tra 12,5% e 50%) e dell\u2019Id-CA (concentrazioni tra 3.12% e 50%) vs i ceppi testati. Inoltre, l'OE e gli Id sono stati analizzati in GC-MS per valutarne la composizione terpenica e, nel caso di M. citriodora, paragonare l\u2019efficacia dell\u2019OE a quella dell\u2019Id in funzione del loro contenuto terpenico. Risultati. L'OE-MC e l\u2019Id-CA sono risultati efficaci, a concentrazioni variabili, su tutti i ceppi testati, mentre, l'Id-MC era attivo su tutti i microrganismi tranne che sui ceppi di P. aeruginosa (MIC>4% v/v). Le analisi GC-MS hanno individuato come principali componenti terpeniche il timolo per OE-MC e di Id-MC (19,6% e 66,4% rispettivamente) e il linalolo per Id-CA. Conclusioni. Nonostante sia necessaria una maggiore concentrazione di Id-MC per ottenere lo stesso effetto inibente dell\u2019OE-MC, la quantit\ue0 di terpeni presenti per unit\ue0 di volume nell\u2019Id \ue8 risultata inferiore rispetto all'EO. La maggiore efficacia dei terpeni contenuti nell\u2019Id \ue8 probabilmente dovuta all'ambiente idrofilo che ne promuove una maggiore disponibilit\ue0

Anti-Mold Effectiveness of a Green Emulsion Based on Citrus aurantium Hydrolate and Cinnamomum zeylanicum Essential Oil for the Modern Paintings Restoration

A modern painting is characterized by multi-material bases extremely exposed to biodeteri-ogenic attacks. The aim of this work was to test the antifungal effectiveness of a natural, eco-friendly, and safe emulsion based on Citrus aurantium L. var. amara hydrolate and Cinnamomum zeylanicum Blume (from bark) essential oil, named “Zeylantium green emulsion” (Zege), on modern paintings. Colored unaged and aged canvas samples, performed with modern techniques (acrylic, vinylic and alkyd), were used to test in vitro both the antifungal effectiveness of Zege and its impact on the chemical–physical characteristics. Microbiological tests were performed according to the EUCAST international guidelines. pH measurements and colorimetric analysis were performed on unaged and aged canvases before and after Zege spray treatment. Finally, in situ tests were performed using the spray emulsion on canvas samples obtained from Ilaria Margutti’s modern artwork, which had been colonized by molds. Microbiological tests on canvas prototypes showed a time-and dose-dependent effectiveness of the Zege spray. None of the techniques underwent relevant changes in pH. Only the acrylic colors were unaffected in the colorimetric analysis, among all colored unaged or aged canvases. Tests made with modern artwork samples confirmed the in situ antifungal effectiveness. The Zege spray showed encouraging results in regard to the use of this formulation in the restoration of modern paintings

Origanum vulgare essential oil vs. A commercial mixture of essential oils: In vitro effectiveness on salmonella spp. from poultry and swine intensive livestock

open11noSalmonella spp. represent a public health concern for humans and animals due to the increase of antibiotic resistances. In this scenario, the use of essential oils (EOs) could be a valid tool against Salmonella contamination of meat. This work compares the in vitro effectiveness of an Italian mixture of feed additives based on EOs (GR-OLI) with EO of Origanum vulgare L., recently admitted by European Food Safety Authority (EFSA) for animal use. Twenty-nine Salmonella serotypes isolated from poultry and pig farms were used to assess GR-OLI and O. vulgare EO antimicrobial propeties. O. vulgare EO was active on the disaggregation of mature biofilm, while GR-OLI was capable of inhibiting biofilm formation and disaggregating preformed biofilm. Furthermore, GR-OLI inhibited bacterial adhesion to Caco-2 cells in a dose-dependent manner. Both products showed inhibition of bacterial growth at all time points tested. Finally, the synergistic action of GR-OLI with commonly used antibiotics against resistant strains was investigated. In conclusion, the mixture could be used both to reduce the meat contamination of Salmonella spp. before slaughter, and in synergy with low doses of ciprofloxacin against resistant strains. Although EOs as feed additives are already used in animal husbandry, no scientific study has ever highlighted their real antimicrobial potential.openDi Vito M.; Cacaci M.; Barbanti L.; Martini C.; Sanguinetti M.; Benvenuti S.; Tosi G.; Fiorentini L.; Scozzoli M.; Bugli F.; Mattarelli P.Di Vito M.; Cacaci M.; Barbanti L.; Martini C.; Sanguinetti M.; Benvenuti S.; Tosi G.; Fiorentini L.; Scozzoli M.; Bugli F.; Mattarelli P

Targeting ddx3x helicase activity with ba103 shows promising therapeutic effects in preclinical glioblastoma models

DDX3X is an ATP-dependent RNA helicase that has recently attracted interest for its involvement in viral replication and oncogenic progression. Starting from hit compounds previously identified by our group, we have designed and synthesized a new series of DDX3X inhibitors that effectively blocked its helicase activity. These new compounds were able to inhibit the proliferation of cell lines from different cancer types, also in DDX3X low-expressing cancer cell lines. According to the absorption, distribution, metabolism, elimination properties, and antitumoral activity, compound BA103 was chosen to be further investigated in glioblastoma models. BA103 determined a significant reduction in the proliferation and migration of U87 and U251 cells, downregulating the oncogenic protein β-catenin. An in vivo evaluation demonstrated that BA103 was able to reach the brain and reduce the tumor growth in xenograft and orthotopic models without evident side effects. This study represents the first demonstration that DDX3X-targeted small molecules are feasible and promising drugs also in glioblastoma

Antibiofilm activity of three different irrigation techniques: An in vitro study

The microbial infection of the endodontic space occurs in a necrotic tooth as a result of dental caries, trauma, periodontal disease, or previous root canal therapy. The disruption of the biofilms and the reduction of the bacterial load inside root canals are crucial for the success of root canal therapy. The aim of this study was to compare, in vitro, the antibiofilm efficacy of a novel passive sonic irrigation (PSI) device with passive ultrasonic irrigation (PUI) and conventional needle irrigation (CNI). Forty-four single-rooted human teeth were inoculated with a culture of E. faecalis for 28 days. The specimens were randomly divided into three groups: PUI, CNI, and PSI (n = 12). The activation protocols were performed using both 17% EDTA and 5.25% NaOCl. Residual bacterial biofilm was taken by means of a canal brush and colony-forming unit (CFU) were counted. The data were analyzed using one-way ANOVA and Games-Howell's post hoc tests. A major reduction in CFU was observed in the PSI and PUI groups, in comparison with the CNI group. No difference was found (p > 0.05) in terms of CFU reduction between PSI and PUI. PSI could be as effective as PUI in the removal of bacterial biofilms from straight root canals

A New Strategy for Glioblastoma Treatment: In Vitro and In Vivo Preclinical Characterization of Si306, a Pyrazolo[3,4-d]Pyrimidine Dual Src/P-Glycoprotein Inhibitor

20siopenOverexpression of P-glycoprotein (P-gp) and other ATP-binding cassette (ABC) transporters in multidrug resistant (MDR) cancer cells is responsible for the reduction of intracellular drug accumulation, thus decreasing the efficacy of chemotherapeutics. P-gp is also found at endothelial cells' membrane of the blood-brain barrier, where it limits drug delivery to central nervous system (CNS) tumors. We have previously developed a set of pyrazolo[3,4-d]pyrimidines and their prodrugs as novel Src tyrosine kinase inhibitors (TKIs), showing a significant activity against CNS tumors in in vivo. Here we investigated the interaction of the most promising pair of drug/prodrug with P-gp at the cellular level. The tested compounds were found to increase the intracellular accumulation of Rho 123, and to enhance the efficacy of paclitaxel in P-gp overexpressing cells. Encouraging pharmacokinetics properties and tolerability in vivo were also observed. Our findings revealed a novel role of pyrazolo[3,4-d]pyrimidines which may be useful for developing a new effective therapy in MDR cancer treatment, particularly against glioblastoma.openFallacara, Anna Lucia; Zamperini, Claudio; Podolski-Renić, Ana; Dinić, Jelena; Stanković, Tijana; Stepanović, Marija; Mancini, Arianna; Rango, Enrico; Iovenitti, Giulia; Molinari, Alessio; Bugli, Francesca; Sanguinetti, Maurizio; Torelli, Riccardo; Martini, Maurizio; Maccari, Laura; Valoti, Massimo; Dreassi, Elena; Botta, Maurizio; Pešić, Milica; Schenone, SilviaFallacara, Anna Lucia; Zamperini, Claudio; Podolski-Renić, Ana; Dinić, Jelena; Stanković, Tijana; Stepanović, Marija; Mancini, Arianna; Rango, Enrico; Iovenitti, Giulia; Molinari, Alessio; Bugli, Francesca; Sanguinetti, Maurizio; Torelli, Riccardo; Martini, Maurizio; Maccari, Laura; Valoti, Massimo; Dreassi, Elena; Botta, Maurizio; Pešić, Milica; Schenone, Silvi