Antimicrobial resistance is a major public health concern worldwide. Albeit to a lesser extent than bacteria, fungi are also becoming increasingly resistant to antifungal drugs. Moreover, due to the small number of antifungal classes, therapy options are limited, complicating the clinical management of mycoses. In this view, antimicrobial peptides (AMPs) are a potential alternative to conventional drugs. Among these, Proline-rich antimicrobial peptides (PrAMPs), almost exclusively of animal origins, are of particular interest due to their peculiar mode of action. In this study, a search for new arginine- and proline-rich peptides from plants has been carried out with a bioinformatic approach by sequence alignment and antimicrobial prediction tools. Two peptide candidates were tested against planktonic cells and biofilms of Candida albicans and Candida glabrata strains, including resistant isolates. These peptides showed similar potent activity, with half-maximal effective concentration values in the micromolar range. In addition, some structural and functional features, revealing peculiar mechanistic behaviors, were investigated.

By-products of the fish industry can be a source of valuable molecules, with different technological applications. Fish bones contain both organic and inorganic phases (collagen/other proteins and calcium phosphate minerals respectively), with several uses and among them in cosmetics and medicine. Using the common extraction protocols, only one of the two phases can be extracted, while the other one is degraded. In the present work a method of extraction which allows the obtainment of both proteins and calcium phosphates was developed. Bones from two different species (salmon and sea bream) were tested. Saline solutions based on (NH4)HCO3/ (NH4)2HPO4 or (NH4)HCO3 were employed for the protein extraction. Analysis of the organic phase showed that the main extracted proteins were collagen in its different forms (especially alpha-1); some differences were observed between salmon and sea bream and between the methods of extraction. As for the mineral phase, after calcination at 700 degrees C, single phase hydroxyapatite was obtained with (NH4)HCO3 solution, while the use of (NH4) HCO3/(NH4)2HPO4 led to a biphasic material of beta-tricalcium phosphate and beta-calcium pyrophosphate. Biocompatibility tests performed on both organic and inorganic extracts (with fibroblast and osteoblast-like cellular lines) showed their non-toxicity and, hence, their potential suitability for cosmetic or biomedical applications. This work shows that it is possible to obtain a more complete valorisation of the fish bones, with the simultaneous extraction of organic and inorganic valuable compounds. The principles of this process could be applied also to bones of other species, with a positive impact on the environment for the reduction of the wastes and a better use of all available resources.

The increased prevalence and incidence of fungal infections, of which Candida albicans represents one of the most life-threatening organisms, is prompting the scientific community to develop novel antifungal molecules. Many essential oils components are attracting attention for their interesting antifungal activities. Given the chemical and physical characteristics of these compounds, the use of appropriate nanodelivery systems is becoming increasingly widespread. In this study, chitosan nanoparticles were prepared using an ionic gelation procedure and loaded with the phenolic monoterpene carvacrol. After a bioassay guided optimization, the best nanoparticle formulation was structurally characterized by means of different spectroscopic (UV, FTIR and DLS) and microscopy techniques (SEM) and described for their functional features (encapsulation efficiency, loading capacity and release kinetics). The antifungal activity of this formulation was assayed with different Candida spp., both in planktonic and biofilm forms. From these studies, it emerged that the carvacrol loaded nanoparticles were particularly active against planktonic forms and that the antibiofilm activity was highly dependent on the species tested, with the C. tropicalis and C. krusei strains resulting as the most susceptible.

Calcium phosphate is a natural biomineral and the major inorganic constituent of bones and teeth. Therefore, synthetic calcium phosphates that mimic the biogenic ones possess excellent biocompatibility as well as biodegradability and are promising materials for medicine. Due to their unique physiochemical properties, calcium phosphate nanoparticles (CaP NPs) are extensively exploited in nano-medicine as carriers of biomolecules, including peptides, proteins, and nucleic acids. In this regard, peptides are of particular interest as they are exceptionally selective and efficacious for the treatment of a broad range of diseases. Among various peptides for biomedical applications, cardio-specific peptides are particularly interesting since they represent a valuable alternative to conventional treatments. Moreover, they can contribute to overcome important clinical limitations, including drug resistance and non-specific biodistribution of traditional drug products. In this work, we have investigated the loading of a therapeutic mimetic peptide, which was previously shown to improve myocardial contraction and results in the restoration of cardiac function. Peptide-loaded CaP NPs were prepared by exploiting a biomineralization approach, by using a mineralizing solution containing Ca, Mg, and PO4 ions. Several experimental conditions were tested by varying the reaction time, as well as the drug concentration. Colloidal stability, morphology, size, as well as drug loading were evaluated to identify the best candidate to be tested in vitro in the future.

The use of inhalable nanoparticles (NPs) for cystic fibrosis (CF) has been advocated as a promising tool to improve the efficacy of antimicrobials taking advantage of their ability to penetrate airway mucus and pathogen biofilm and to release the drug in or in proximity to the enclosed bacteria. Here, inhalable calcium phosphate (CaP) NPs were functionalized with colistin (Col) which is one of the most active antimicrobials against Gram-negative bacteria. The adsorption kinetic and isotherm of Col on CaP-NPs were investigated and fitted according to different mathematical models and revealed an electrostatic interaction between positively charged amine groups of Col and negatively charged surface of CaP-NPs. The maximum Col payload was of about 50 mg g of CaP-NPs. After functionalization, despite an increase of size (213 vs 95 nm), in citrate solution, CaP-NPs maintained a dimension and surface charge considered suitable for crossing mucus barrier. CaP-NPs do not interact with mucin and are able to permeate a layer of artificial mucus. In vitro tests on pulmonary cells demonstrated that CaP-NPs are not cytotoxic up to a concentration of 125 ?g mL. The antimicrobial and antibiofilm activity of Col loaded CaP-NPs tested on Pseudomonas aeruginosa RP73, a clinical strain isolated from a CF patient, was similar to that of free Col demonstrating that the therapeutic effect of Col adsorbed on CaP-NPs was retained. This work represents the first attempt to use CaP-NPs as delivery system for the CF treatment. The encouraging results open the way to further studies.

By products of the fish industry can be a source of valuable molecules, with different technological applications. Fish bones, in particular, contain both organic and inorganic phases (collagen, other proteins and calcium phosphate respectively), with uses in cosmetics and biomedicine. Using the most common extraction protocols, only of the two phases can be extracted, while the other one is degraded. Investigation of the organic phase composition by proteomic related procedures followed by LC MS MS analysis, showed that the main extracted proteins were collagen in its different forms (especially alpha 1); some differences were observed between salmon and sea bream and between the methods of extraction. Biocompatibility tests performed on both organic and inorganic extracts (with fibroblast and osteoblast like cellular lines) showed their non toxicity and, hence, their suitability for cosmetic or biomedical applications.

The purpose of this study was to correlate the chemical composition of four commercial concentrated glycerine macerates (C-GMs), produced through the same extraction method, with their in vitro antimicrobial, antioxidant, and immunomodulatory properties, in order to evaluate their potential for healing upper airway diseases. C-GMs of Carpinus betulus (CB), Ficus carica (FC), Alnus glutinosa (AG) and Ribes nigrum (RN) were studied. The quality was evaluated using HPLC and IM-SPME/GC-MS systems; anti-oxidant and anti-microbial activities were assessed by the respective DPPH test, and micro-broth dilution test performed against 10 strains of Streptococcus pyogenes and 10 probiotic strains. ELISA and MTT tests were used to assess the immunomodulatory activity and the cytotoxicity of C-GMs, respectively. A significant correlation was found between the number of active compounds and the in vitro C-GMs effectiveness. Furthermore, the C-GMs of AG showed the best anti-microbial activity on pathological strains and, together with CB, the best anti-oxidant activity. The ELISA test exhibited a good immunomodulatory activity of RN. In vitro data support the integrated use of C-GMs of CB, AG, and RN in presence of airway diseases, and highlight the importance of standard procedures in cultivation, harvest and post-harvest treatments, as a premise for C-GMs with consistent characteristics.

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Background: Irritable bowel syndrome (IBS) is a functional disorder without any pathological alteration, in which the alterations of the Candida/Saccharomyces ratio of the gut microbiota, the balance of pro and anti-inflammatory cytokines and the brain-gut-microbiome axis are important for the development and progression of IBS. The aim of the study was to identify natural products, including essential oils or hydrolates, which were contextually harmless for the gut beneficial strains (e.g. Saccharomyces spp.) but inhibitory for the pathogenic ones (Candida spp.). Methods: The effectiveness of 6 essential oils and 2 hydrolates was evaluated using microbiological tests, carried out on 50 clinical isolates (Candida, Saccharomyces and Galattomyces species) and 9 probiotic strains (Saccharomyces cerevisiae, Lactobacillus species, Akkermansia muciniphila and Faecalibacterium prausnitzii) and immunological and antioxidant assays. Results: The study led to a mixture based on a 1/100 ratio of Citrus aurantium var. amara essential oil/Vitis vinifera cv Italia hydrolate able to contextually reduce, in a concentration-dependent manner, the ability of Candida species to form hyphal filaments and have an interesting immunomodulatory and anti-oxidant action. This mixture can potentially be useful in the IBS treatment promoting the restoration of the intestinal microbial and immunological balance.

In the modern view of selective drug delivery of bioactive molecules, the attention is moving onto the setup of the perfect carrier more than in the optimization of the active compound. In this respect, virus-like particles constitute bioinspired nanodevices with the intrinsic ability to transport a large class of molecules, ranging from smart drugs to small interfering RNAs. In this work, we demonstrate the efficacy of a novel construct obtained by fusing a self-assembling protein from the human Rotavirus A, VP6, with the Small Ubiquitin Modifier domain, which maintains the ability to form nanoparticles and nanotubes and is able to be used as a drug carrier, even without specific targeting epitopes. The high expression and purification yield, combined with low toxicity of the empty particles, clearly indicate a good candidate for future studies of selective drug delivery. © 2018 American Institute of Chemical Engineers Biotechnol. Prog., 35: e2769, 2019.

Cystic fibrosis (CF) is a genetic disease affecting today nearly 70,000 patients worldwide and characterized by a hypersecretion of thick mucus difficult to clear arising from the defective CFTR protein. The over-production of the mucus secreted in the lungs, along with its altered composition and consistency, results in airway obstruction that makes the lungs susceptible to recurrent and persistent bacterial infections and endobronchial chronic inflammation, which are considered the primary cause of bronchiectasis, respiratory failure, and consequent death of patients. Despite the difficulty of treating the continuous infections caused by pathogens in CF patients, various strategies focused on the symptomatic therapy have been developed during the last few decades, showing significant positive impact on prognosis. Moreover, nowadays, the discovery of CFTR modulators as well as the development of gene therapy have provided new opportunity to treat CF. However, the lack of effective methods for delivery and especially targeted delivery of therapeutics specifically to lung tissues and cells limits the efficiency of the treatments. Nanomedicine represents an extraordinary opportunity for the improvement of current therapies and for the development of innovative treatment options for CF previously considered hard or impossible to treat. Due to the peculiar environment in which the therapies have to operate characterized by several biological barriers (pulmonary tract, mucus, epithelia, bacterial biofilm) the use of nanotechnologies to improve and enhance drug delivery or gene therapies is an extremely promising way to be pursued. The aim of this review is to revise the currently used treatments and to outline the most recent progresses about the use of nanotechnology for the management of CF.

One of the main prerequisite of a therapeutic drug is to overcome a series of physiological barriers and it to be less toxic for the human. To this aim, it is necessary to find a carrying vector able to enhance drug internalization and to improve its therapeutic efficacy. Presently, biocompatible and biodegradable nanoparticles (NPs) as some effective drug delivery devices, are widely studied. In order to increase the antimicrobial and antitumor efficacy of different natural products numerous studies are in progress to evaluate different drug delivery systems (cationic liposomes, lysozyme-shelled hollow nanoimicrobubbles and nanoimicrocapsules). This delivery system would have a synergistic and additive effect decreasing the drug resistance reactions too. The present article has tried to review some of the latest research on nanocarriers in clinical care.

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Proteomic surveys with top-down platforms are today revealing thousands of naturally occurring fragments of bigger proteins. Some of them have not functional meaning because they derive from pathways responsible for protein degradation, but many have specific functions, often completely different from that one of the parent proteins. These peptides encrypted in the protein sequence are nowadays called cryptides. They are frequent in the animal and plant kingdoms and represent a new interesting -omic field of investigation. To point out how much widespread is their presence, we describe here the most studied cryptides from very common sources such as serum albumin, immunoglobulins, hemoglobin, and from saliva and milk proteins. Given its vastness, it is unfeasible to cover the topic exhaustively, therefore only several selected examples of cryptides from other sources are thereafter reported. Demanding is the development of new -omic platforms for the functional screening of new cryptides, which could provide suggestion for peptides and peptido-mimetics with variegate fields of application.

Marine environment is acquiring even more interest as a source of new bioactive compounds, among these marine organism's derived bioactive peptides are considered a promising group of natural substances exhibiting different biological activities: antimicrobial, anticancer, antihypertensive, anti-inflammatory and so on. In particular antimicrobial activity may encounter the emerging severe problem of the antibiotic-resistant microorganisms. Among marine animals, sponges have attracted in the years a great interest of pharmacologists, chemists and biochemists as a rich source of peculiar antimicrobial compounds that they have evolved to protect themselves due to their sessile nature. Sponges produce a great variety of antimicrobial peptides with peculiar structural features showing antifungal, antibacterial and antiviral properties. In this review we will focus on those peptides showing the most interesting antimicrobial activities as a proposal for their exploitation as new future drugs

Methicillin-resistant Staphylococcus aureus (MRSA) is responsible for serious hospital infections worldwide and represents a global public health problem. Curcumin, the major constituent of turmeric, is effective against MRSA but only at cytotoxic concentrations or in combination with antibiotics. The major issue in curcumin-based therapies is the poor solubility of this hydrophobic compound and the cytotoxicity at high doses. In this paper, we describe the efficacy of a composite nanoparticle made of curcumin (CU) and graphene oxide (GO), hereafter GOCU, in MRSA infection treatment. GO is a nanomaterial with a large surface area and high drug-loading capacity. GO has also antibacterial properties due mainly to a mechanical cutting of the bacterial membranes. For this physical mechanism of action, microorganisms are unlikely to develop resistance against this nanomaterial. In this work, we report the capacity of GO to support and stabilize curcumin molecules in a water environment and we demonstrate the efficacy of GOCU against MRSA at a concentration below 2 mg ml21. Further, GOCU displays low toxicity on fibroblasts cells and avoids haemolysis of red blood cells. Our results indicate that GOCU is a promising nanomaterial against antibiotic-resistant MRSA.

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