Search

The fresh bulbs of Stenomesson miniatum, a plant belonging to Amaryllidaceae family with a poorly investigated phytochemical profile, were traditionally employed by Andean healers to treat tumors and abscesses. The aims of this study were to characterize the alkaloid extract from the bulbs of S. miniatum and test its cytotoxic and antibacterial potential. The alkaloid extract was characterized by dereplication using various techniques (CPC, NMR, UPLC-HRMS) and referring to in-home or online databases for spectroscopic data matching. Cytotoxic activities were evaluated on A431 human epidermoid carcinoma cells through a metabolic assay, and on Jurkat human acute T-leukemia cells through a cell-impermeant fluorescent nuclear probe. Antibacterial assays were carried out against Staphylococcus aureus, Staphylococcus epidermidis and Streptococcus pyogenes by using a standardized microdilution broth method 96-well plate. Eleven known Amaryllidaceae alkaloids were identified together with another compound determined as being an extraction artefact. The alkaloid extract showed good cytotoxic activity against both the tumor cell lines, reaching an IC50 of 3.3 µg/mL against A431 cells and of 10.9 µg/mL against Jurkat cells. The analysis of the fractions allowed the activity to be attributed to the presence of pretazettine and haemathamine. Conversely, no antibacterial activities were recorded for any of the samples.

The development of infections still represents a severe problem in orthopedic field. In this work, we developed an anti-microbial and radiopaque calcium phosphate bone cement with a long-lasting inhibitory activity thanks to the addition of gentamicin sulphate-loaded solid lipid microparticles obtained by spray congealing. Cements containing gentamicin both added into cement powder and loaded into microparticles were prepared and characterized and their properties were compared. The results of structural, morphological, mechanical and in vitro characterizations indicate that inclusion of gentamicin into microparticles is a good approach in order to obtain materials able to display a strong inhibition towards the growth of Gram-negative, as well as Gram-positive bacteria and to enhance the viability of osteoblast-like cells. All the compositions have an excellent activity also against clinical isolates, including multi drug resistant phenotypes. In particular, cements containing gentamicin-loaded microparticles display a strong and long-term inhibitory activity. Suitable modification of the liquid/powder ratio provided injectable formulation, whereas addition of BaSO4 produced radiopacity. All the formulations display good injectability and cohesion, and no evidence of demixing.

Clofazimine (CLZ) is an antibiotic with a promising behavior against Gram-positive bacteria; however, the drug is completely insoluble in water and accumulates in fat tissues. We explored nanocarriers, labeled and not labeled with rhodamine, consisting of negatively charged sulfobutylether-beta-cyclodextrins for CLZ loading. A new oligomeric carrier was obtained cross-linking beta CyD with epichlorohydrin followed by sulfonation in a strongly alkaline aqueous medium. The oligomeric carrier has a MW of 53 kDa and forms small nanoparticles of a few tens of nm. With aqueous solutions containing a 25 mg/mL oligomeric carrier, we loaded up to 0.5 mg/mL of drug. The oligomers exhibited a 10-fold better loading capacity compared to monomers and formed nanoparticles with a size in the 20-60 nm range after drug loading. Circular dichroism confirmed encapsulation of the CLZ in the nanocarriers. All carriers with or without CLZ are not cytotoxic up to 1 mu M, while CLZ alone is highly cytotoxic at the same concentration. The drug has IC50 values below 100 nM against S. epidermidis. The same holds true also for clinical isolates of S. epidermidis, some displaying MDR So, the selectivity index significantly increased for CLZ/carrier systems compared to the drug alone. Taken all together, our results open new avenues for the clinical application of this antibiotic.

Summary Candida albicans and Staphylococcus aureus are common human pathogens , frequently isolated independently or co-isolated from bloodstream infections, and able to form dense polymicrobial biofilms on various medical devices resulting in strong resistance to conventionally used antimicrobials. New and innovative approaches are therefore needed to ensure the successful management of biofilm related infections. In this study, a chalcone-based derivative and a polycyclic anthracene-maleimide adduct, previously ascertained by us as inhibitors of C. albicans and S. aureus growths, respectively, were reconsidered in a new perspective by evaluating the efficacy of a combined treatment against a polymicrobial biofilm. Both quantitative and qualitative analyses were carried out to delve into their inhibitory potential on the polymicrobial population. Our results indicate that these newly identified antimicrobials are effective in reducing the biomass of the mixed C. albicans-S. aureus biofilm and the viability of fungal-bacterial cells within the polymicrobial community; in addition, confocal laser scanning microscopy demonstrates that the combined treatment thoroughly modifies the architecture of the dual-species biofilm.

In recent decades, synthetic plastic polymers have been the most practical and economical solution for packaging applications due to their low cost, availability, excellent optical, mechanical and barrier properties and resistance against water. However, most of the plastics used for packaging are hardly biodegradable. With a view to a circular economy, the aim of this work focused on the development of a new material made of commercial cellulose derivatives (hydroxypropyl methyl cellulose or carboxymethyl cellulose) mixed with snail mucus extracted from Helix Aspersa Muller. Increasing in Snail Mucus content enhances films elongation and adhesion strength while decreasing water vapor permeability. The cellulose-snail mucus based films are highly transparent but, more interestingly, the mucus confers UV-screening effect. In addition, the composite films exhibit antimicrobial activity against both Gram-positive and Gram-negative bacteria. Furthermore, snail mucus addition to carboxymethyl cellulose strongly decreases films solubility in water. The biodegradation tests indicate that all the films degrade in soil between two and four weeks. The excellent results indicate that these biocomposite films are very good candidate for food packaging.

Lactic acid bacteria (LAB) &ldquo, fermentates&rdquo, confer a beneficial effect on intestinal function. However, the ability of new fermentations to improve LAB broth activity in preventing pathogen-induced intestinal inflammation and barrier dysfunction has not yet been studied. The objective of this study was to determine if broths of LAB fermented with Eruca sativa or Barbarea verna seed extracts prevent gut barrier dysfunction and interleukin-8 (CXCL8) release in vitro in human intestinal Caco-2 cells infected with enterohemorrhagic Escherichia coli (EHEC) O157:H7. LAB broths were assayed for their effects on EHEC growth and on Caco-2 viability, thereafter, their biological properties were analysed in a co-culture system consisting of EHEC and Caco-2 cells. Caco-2 cells infected with EHEC significantly increased CXCL8 release, and decreased Trans-Epithelial Electrical Resistance (TEER), a barrier-integrity marker. Notably, when Caco-2 cells were treated with LAB broth enriched with E. sativa seed extract and thereafter infected, both CXCL8 expression and epithelial dysfunction reduced compared to in untreated cells. These results underline the beneficial effect of broths from LAB fermented with E. sativa seed extracts in gut barrier and inflammation after EHEC infection and reveal that these LAB broths can be used as functional bioactive compounds to regulate intestinal function.

Several strategies have been explored to obtain effective econazole nitrate (ECN) concentrations at the site of application for a prolonged time. In this paper, different gelatin-based film formulations for vaginal application were investigated, containing ECN (10% w/w with respect to gelatin) as pure drug or as drug-solid dispersions (SD). For the production of SD, different polymers were evaluated: polyvinylpyrrolidone (PVP), Soluplus® (polyvinyl caprolactam-polyvinyl acetate-polyethylene glycol graft copolymer) and Gelucire® 50/13 (mixture of mono-, di- and triglycerides of fatty acids, esters of PEG 1500 and free PEG). Gelucire®-SD showed the best solubility enhancement, increasing 9.2 times the ECN solubility in pH 4.5 solution respect to pure drug; DSC and XRD analysis confirmed the crystalline form of the drug. XRD results evidenced that all gelatin-based films, containing either the drug or the SD, underwent the topotactic transformation of ECN into crystalline econazole (EC), owing to a strong interaction between the drug and the gelatin. Films containing Gelucire®-based SD displayed lower brittleness and rigidity with respect to the other samples; moreover they demonstrated good structural integrity after 24 h of incubation in the acidic solution (swelling degree of about 350%). Then, Gelucire®-SD based films were compared with the corresponding formulations cross-linked by genipin (2% w/w). The addition of genipin did not interfere with the drug-gelatin interaction. Gelucire®-SD based films showed similar release profiles to neat gelatin films, enhancing the drug release in the first 5 h and controlling the EC release over time, avoiding the use of a crosslinking additive. Finally, gelatin films containing Gelucire® solid dispersion displayed good adhesiveness and anti-Candida activity. Overall, results support the potential use of this film formulation as noncytotoxic EC delivery system for the treatment of vaginal candidiasis.

Extensive documentation is available on plant essential oils as a potential source of antimicrobials, including natural drugs against Candida spp. Yeasts of the genus Candida are responsible for various clinical manifestations, from mucocutaneous overgrowth to bloodstream infections, whose incidence and mortality rates are increasing because of the expanding population of immunocompromised patients. In the last decade, although C. albicans is still regarded as the most common species, epidemiological data reveal that the global distribution of Candida spp. has changed, and non-albicans species of Candida are being increasingly isolated worldwide. The present study aimed to review the anti-Candida activity of essential oils collected from 100 species of the Lamiaceae family growing in the Mediterranean area and the Middle East. An overview is given on the most promising essential oils and constituents inhibiting Candida spp. growth, with a particular focus for those natural products able to reduce the expression of virulence factors, such as yeast-hyphal transition and biofilm formation. Based on current knowledge on members of the Lamiaceae family, future recommendations to strengthen the value of these essential oils as antimicrobial agents include pathogen selection, with an extension towards the new emerging Candida spp. and toxicological screening, as it cannot be taken for granted that plant-derived products are void of potential toxic and/or carcinogenic properties.

The large amount of cauliflower industry waste represents an unexplored source of bioactive compounds. In this work, peptide hydrolysates from cauliflower leaves were characterized by combined bioanalytical approaches. Twelve peptide fractions were studied to evaluate unexplored biological activities by effect-based cellular bioassays. A potent inhibition of intracellular xanthine oxidase activity was observed in human vascular endothelial cells treated with one fraction, with an IC50 = 8.3±0.6 μg/ml. A different fraction significantly induced the antioxidant enzyme superoxide dismutase 1 and decreased the tumor necrosis factorα-induced VCAM-1 expression, thus leading to a significant improvement in the viability of human vascular endothelial cells. Shotgun peptidomics and bioinformatics were used to retrieve the most probable bioactive peptide sequences. Our study shows that peptides from cauliflower waste should be recycled for producing valuable products useful for the prevention of endothelial dysfunction linked to atherogenesis progression.

In this work we developed new antibacterial composite materials using polydopamine (PDA) to trigger the deposition of silver nanoparticles (AgNPs) onto calcium phosphates, namely octacalcium phosphate (OCP) and Alpha-tricalcium phosphate (TCP). Functionalization of OCP and TCP with a self-polymerized polydopamine layer was obtained by soaking the calcium phosphates in dopamine solution. The PDA surface of functionalized calcium phosphates (OCPd and TCPd) promoted the deposition of AgNPs by reducing silver ions when soaked in a silver nitrate solution. The amount of deposited AgNPs can be modulated by varying the concentration of silver nitrate solution and the type of substrate. The results of in vitro tests carried out with osteoblast-like MG63 cells indicate that the combination of AgNPs with OCP provides more biocompatible materials than those obtained using TCP as substrate. In particular, the study of osteoblast activity and differentiation was focused on the samples OCPdAg5 (silver content = 8.2 wt%) and TCPdAg5 (silver content = 4.7 wt%), which did not show any cytotoxicity, and compared with those obtained on pure OCP and TCP. The results demonstrate that the AgNPs loaded materials support osteoblast viability and differentiation, whereas they significantly inhibit the growth of relevant antibiotic-resistant pathogenic bacteria.

Snail mucus is an attractive natural substance, which is increasingly used in cosmetic creams and syrups thanks to its emollient, moisturizing, protective and reparative properties. The aim of the present study was to explore the physicochemical properties of chitosan-based films added with snail mucus extracted from Helix Aspersa Muller. To this aim, chitosan films at different content of snail mucus were fabricated by simple solvent casting technique. The results of X-ray diffraction analyses, tensile mechanical tests, Infrared spectroscopy and thermogravimetry demonstrated that snail mucus addition strongly modifies the properties of chitosan films. In particular, it acted like a plasticizer enhancing films extensibility up to ten times and strongly improving their water barrier and bioadhesion properties, with a trend depending on Snail mucus content. Furthermore, it provides the films with antibacterial properties and enhanced cytocompatibility, yielding materials with tailored properties for specific requirements.

Escherichia coli may innocuously colonize the intestine of healthy subjects or may instigate infections in the gut or in other districts. This study investigated intestinal E. coli isolated from 20 healthy adults. Fifty-one strains were genotyped by molecular fingerprinting and analyzed for genetic and phenotypic traits, encompassing the profile of antibiotic resistance, biofilm production, the presence of surface structures (such as curli and cellulose), and their performance as recipients in conjugation experiments. A phylogroup classification and analysis of 34 virulence determinants, together with genes associated to the pks island (polyketide-peptide genotoxin colibactin) and conjugative elements, was performed. Most of the strains belonged to the phylogroups B1 and B2. The different phylogroups were separated in a principal coordinate space, considering both genetic and functional features, but not considering pulsed-field gel electrophoresis. Within the B2 and F strains, 12 shared the pattern of virulence genes with potential uropathogens. Forty-nine strains were sensitive to all the tested antibiotics. Strains similar to the potential pathogens innocuously inhabited the gut of healthy subjects. However, they may potentially act as etiologic agents of extra-intestinal infections and are susceptible to a wide range of antibiotics. Nevertheless, there is still the possibility to control infections with antibiotic therapy.

In this paper, a small series of anthracene-maleimide-based compounds was prepared and evaluated to assess the antimicrobial potential of this polycyclic core, a scaffold previously unexplored for new antibiotic development. Some of the new compounds showed appreciable anti

Gelatin-based films enriched with snail slime are proposed as novel biodegradable and naturally bioadhesive patches for cutaneous drug delivery. Films (thickness range 163–248 μm) were stretchable and they adhered firmly onto the wetted skin, especially those with high amount (70% V/V) of snail slime extract. Fluconazole was selected as model drug and added to films containing the highest amount of snail slime. The presence of Fluconazole (4.53 ± 0.07% w/w) did not modify significantly the mechanical properties, the swelling degree and the bioadhesive performances of the films. Structural investigations demonstrated that the crystalline form III of the drug changed to the amorphous one, forming an amorphous solid dispersion. Moreover, snail slime prevented the drug recrystallization over time. In vitro permeation studies showed that film exhibited a cumulative drug concentration (over 60% in 24 h) similar to that of the control solution containing 20% w/V of ethanol. Fluconazole-loaded gelatin films proved to be effective towards clinical isolates of Candida spp. indicating that the drug maintained its remarkable antifungal activity once formulated into gelatin and snail slime-based films. In conclusion, snail slime, thanks to its peculiar composition, has proved to be responsible of optimal skin adhesion, film flexibility and of the formation of a supersaturating drug delivery system able to increase skin permeation.

Parvovirus B19 (B19V) is a human ssDNA virus responsible for a wide range of clinical manifestations, still lacking for a specific antiviral therapy. The identification of compounds active against B19V may add therapeutic options to the treatment of B19V infections, that now entirely relies on symptomatic treatments. In the search for compounds possibly inhibiting B19V replication, a particular focus was raised to cidofovir, an acyclic nucleoside phosphonate broadly active against dsDNA viruses. The present study was aimed at evaluating the effect of cidofovir against B19V in two model systems, the UT7/EpoS1 cell line and erythroid progenitor cells (EPC), generated from peripheral blood mononuclear cells. Experiments were carried out at different multiplicity of infections and cidofovir concentrations (0-500 μM) during a course of infection. The effects of cidofovir on B19V replication were assessed by qPCR assays while influence of cidofovir on host cells was measured by cell proliferation and viability assays. Our findings demonstrated that cidofovir has a relevant inhibiting activity on B19V replication within infected UT7/EpoS1, and that the effect on B19V DNA amounts is dose-dependent allowing for the determination of EC50 and EC90 values (7.45-41.27 μM, and 84.73-360.7 μM, respectively). In EPCs, that constitute a cellular population close to the natural target cells in bone marrow, the inhibitory effect was demonstrated to a lesser extent, however provoking a significant reduction on B19V DNA amounts at 500 μM (68.2-92.8%). To test infectivity of virus released from EPCs cultured in the presence of cidofovir, cell culture supernatants were used as inoculum for a further course of infection in UT7/EpoS1 cells, indicating a significant reduction in viral infectivity at 500 μM cidofovir. Since the drug did not interfere with the overall cellular DNA synthesis and metabolic activity, the observed effect of cidofovir could be likely related to a specific inhibition of B19V replication.

The aim of this study was to determine the in vitro antimicrobial activity of crude extracts of three plants from Ayurveda tradition (Tinospora cordifolia, Alstonia scholaris, Crataeva nurvala) against reference microbial strains and clinical isolates of methicillin-resistant Staphylococcus aureus (MRSA) and carbapenemase-producing Klebsiella pneumoniae. IC50 values were obtained by micro-dilution methods meeting the requirements of the NCCLS standard. The cytotoxicity of the extracts was also investigated on a mammalian cell line. Extracts displayed a variable degree of antimicrobial activity and did not interfere with mammalian cell proliferation. T. cordifolia and A. scholaris exhibited a higher inhibitory activity against clinical isolates of MRSA and carbapenemase-producing K. pneumoniae compared with reference strains, while C. nurvala exhibited a different behaviour. An antifungal activity towards Candida albicans was observed for A. scholaris extract. Results indicate that constituents from T. cordifolia and A. scholaris may be a potential source of new therapeutic strategies for infectious diseases.

Quercetin, a flavonoid widely diffused in fruits and vegetables, is known for its good pharmacological qualities, such as anti-oxidant and anti-inflammatory properties. In this work, we loaded quercetin on gelatin films with the aim to develop materials with tailored anti-oxidant, mechanical and stability properties. To this purpose, gelatin films at increasing flavonoid content were prepared using two different solvents, namely H2O/EtOH (EtOH films) and DMSO (DMSO films). Quercetin content increased up to about 3.8 and 1.8 wt% in DMSO and EtOH films, respectively. The use of DMSO as solvent prevents the partial regain of collagen triple helix structure during gelling of gelatin sols and results in remarkable extensibility of the films. At variance, EtOH films display X-ray diffraction patterns and DSC plots in agreement with the presence of triple helix structure, and exhibit reduced swelling and increasing mechanical properties on increasing quercetin content. Moreover, their values of denaturation enthalpy indicate the presence of chemical interaction between the flavonoid and gelatin, which can be responsible of their lower quercetin release in PBS in comparison to DMSO films. The flavonoid release is sustained for both series of films and occurs through anchorage to gelatin nanoparticles. Moreover, both DMSO and EtOH functionalized films exhibit relevant anti-oxidant properties, in agreement with their RSA levels, which are comparable to that of pure quercetin.

Parvovirus B19 (B19V), a single-stranded DNA virus in the family Parvoviridae, is a human pathogenic virus responsible for a wide range of clinical manifestations. Currently there is no approved antiviral therapy for parvovirus infection. The acyclic nucleoside phosphonate cidofovir (CDV) has been demonstrated to inhibit replication of B19V in vitro. The aim of the present study was to evaluate whether brincidofovir (BCV), a novel lipid conjugate of CDV, could also inhibit B19V replication. Experiments were carried out in erythroid progenitor cells (EPCs) and UT7/EpoS1 cells, infected with B19V and cultured in the presence of different concentrations of BCV and CDV for comparison. The dynamics of viral replication was evaluated by a qPCR-based assay and the extent of inhibition of viral replication exerted by the compounds determined, along with the effect of the compounds on cell viability and cell proliferation rates. Results confirmed that BCV showed significantly higher antiviral activity against B19V compared to CDV in both cell-based systems. For BCV, the calculated EC50 values were in the range 6.6–14.3 μM in EPCs and 0.22–0.63 μM in UT7/EpoS1 cells. In comparison, the EC50 values for CDV were >300 μM in EPCs and 16.1 μM in UT7/EpoS1 cells. Concurrently, the effects on cell viability were observed at a much higher concentration of BCV, with calculated CC50 values in the range 93.4–102.9 μM in EPCs and 59.9–66.8 μM in UT7/Epos1. The antiviral activity was observed specifically with the metabolically active stereoisomer of BCV suggesting that CDV-diphosphate, the metabolite of both BCV and CDV, was the active antiviral. Our results support a selective role for BCV in the inhibition of B19 viral replication.

In this paper, thirty-six extracts from Sardinian plants were evaluated in vitro for their antimicrobial activity towards a panel of reference strains, Staphylococcus aureus, S. epidermidis, Klebsiella pneumoniae and Escherichia coli, and for their cytotoxicity on mammalian cells. The biological data, together with total phenolic and flavonoid content of the extracts, were treated by PCA (Principal Components Analysis), which highlighted the positive correlation among total phenolic content and increasing antibacterial activities, and a possible involvement of flavonoids in mitigate the cytotoxicity. Thirteen extracts displayed relevant IC50 values (half maximal inhibitory concentration) on S. aureus (IC50 from 1.4 to 153.6 μg/mL), ten out of them were active also against S. epidermidis (IC50 from 3.9 to 150 μg/mL), seven against K. pneumoniae (IC50 from 28.5 to 97.5 μg/mL), and two against E. coli (IC50 74.9 and 156.3 μg/mL). In particular, three extracts obtained from Pistacia terebinthus ssp. terebinthus, Cytinus hypocistis and Limonium morisianum emerged as promising antibacterial candidates. They exhibited remarkable inhibitory activity towards bacterial strains from clinical specimens and presenting different antibiotic-resistance profiles.

Clofazimine (CLZ) is an antibiotic with a promising behavior against Gram-positive bacteria; however, the drug is completely insoluble in water and accumulates in fat tissues. We explored nanocarriers, labeled and not labeled with rhodamine, consisting of negatively charged sulfobutylether-β-cyclodextrins for CLZ loading. A new oligomeric carrier was obtained cross-linking βCyD with epichlorohydrin followed by sulfonation in a strongly alkaline aqueous medium. The oligomeric carrier has a MW of 53 kDa and forms small nanoparticles of a few tens of nm. With aqueous solutions containing a 25 mg/mL oligomeric carrier, we loaded up to 0.5 mg/mL of drug. The oligomers exhibited a 10-fold better loading capacity compared to monomers and formed nanoparticles with a size in the 20-60 nm range after drug loading. Circular dichroism confirmed encapsulation of the CLZ in the nanocarriers. All carriers with or without CLZ are not cytotoxic up to 1 μM, while CLZ alone is highly cytotoxic at the same concentration. The drug has IC

The phytochemical profile, antioxidant activity and α-glucosidase inhibitory activity of Hypericum scruglii Bacchetta, Brullo et Salmeri and Hypericum hircinum L. were analyzed and compared to those of the best-known Hypericum perforatum L. Samples were prepared using three different extraction methods (maceration in 70% ethanol, extraction in MeOH/phosphate buffer and decoction), the influence of which on the metabolic profile and bioactivities are discussed. H. scruglii and H. hircinum extracted in 70% ethanol strongly inhibited α-glucosidase (IC50 7.25 and 14.05 μg/ml, respectively), which is a valuable enzymatic target for treating metabolic disorders, while H. perforatum was found to be less powerful than the other two species in all the performed biological tests. The phytochemical profile was analyzed by NMR, HPLC-DAD and HPLC-FLD, revealing remarkable differences among the species. In particular, H. scruglii, which is a species endemic to Sardinia Island (Italy), was particularly enriched in two phloroglucinol-derivatives identified by means of 2D NMR and LC/MS/MS experiments as 3-geranyl-1-(2′-methylbutanoyl)-phloroglucinol and 3-geranyl-1-(2′-methylpropanoyl)-phloroglucinol.

The present paper describes the development of an integrated lab-on-chip, in which viral DNA amplification with real-time on-chip detection is carried out under constant temperature of 65 °C. The lab-on-chip is composed of a disposable 10-μL polydimethylsiloxane reaction chamber which is thermally and optically coupled to a glass substrate that hosts a thin-film metallic resistive heater and thin-film amorphous silicon diodes which act as temperature and radiation sensors. A loop-mediated isothermal amplification (LAMP) technique was optimized to specifically amplify parvovirus B19 DNA and coupled with Bioluminescent Assay in Real Time (BART) technology to provide real-time detection of target DNA. The experimental results demonstrate the ability of the proposed device to discriminate among different concentrations of viral DNA with an excellent agreement with standard off-chip methods.

The development of new biomaterials able to favor bone formation and to inhibit bone abnormal resorption is mandatory to face the increasing number of age-related musculo-skeletal disorders. Moreover, the increasing antibiotic resistance of clinically important bacteria, which is among the main causes of implant failure, requires new antimicrobial systems. In this study, we prepared multifunctional materials consisting of hydroxyapatite-zoledronate composite crystals decorated with Ag Nanoparticles (AgNPs). Zoledronate, a potent bisphosphonate widely applied for the treatment of pathologies associated to abnormal bone loss, was incorporated into hydroxyapatite up to about 8 wt%. Loading of poly(ethylenimine) – stabilized AgNPs onto the crystals was promoted by zoledronate functionalization and provoked a significant variation of the values of zeta potential. The results of in vitro tests demonstrate that the multifunctional materials combine the beneficial actions of zoledronate and AgNPs. In fact, they improve osteoblast differentiation and activity, whereas they inhibit osteoclastogenesis and osteoclast differentiation, and significantly hinder the growth of multi-drug resistant Gram positive and Gram negative bacteria. As a consequence, they can be exploited both as antiresorptive agents and as antimicrobial materials able to prevent the development of bone-associated infections.

Parvovirus B19 (B19V) is a human pathogenic virus associated with a wide range of clinical conditions. In pregnancy, B19V poses a potential hazard to the fetus as crossing the placental barrier and infecting erythroid progenitor cells in bone marrow and liver, it blocks fetal erythropoiesis leading to profound anemia, hydrops and/or fetal death. The virus is not regarded as a teratogen, however more scientific awareness is emerging on mechanisms and consequences of intrauterine infection and possible sequelae in the neonatal development. Reliable diagnostic procedures and fetal management strategies, including intrauterine transfusion, are established. In spite of being a recognized fetotropic agent possibly leading to fetal loss, testing for B19V is not routinely included in preconception or antenatal screenings, possibly delaying the management of B19V-complicated pregnancies. Continuous advances in B19V research will provide for better diagnostic methods and algorithms, as well as for the development of effective prophylactic interventions and novel therapeutic options.

Parvovirus B19 (B19V), a human pathogenic ssDNA virus in the Parvoviridae family, is characterized by a strict tropism towards erythroid progenitor cells in bone marrow. Infections commonly induce self-limiting clinical manifestations, as erythema infectiosum in children and arthropathies mainly in adults, but, in subjects with underlying hematological disorders or with immune system deficits, the viral-induced arrest of erythropoiesis can lead to severe clinical manifestations requiring hospitalization and supportive transfusion or passive immunization treatments.

Human parvovirus B19 (B19V) is a single-stranded DNA virus. The genome encodes a multifunctional non-structural protein (NS), two capsid proteins (VP1, VP2) and other small non-structural proteins (7.5 kDa, 9 kDa, 11 kDa). Within sensitive cells, B19V can achieve a productive replicative cycle or, on the contrary, establish persistence; differences in its expression profile have been yet investigated following in vitro infections by methodologies enabling information on the entire infected cell population. Conversely, the present study reports quantitative data on the production of B19V analytes (DNA, RNAs, and proteins) at single cell-level, underlining cell-to-cell differences through the viral specific macromolecular synthesis process. Microscope imaging assays (in situ hybridization and immunocytochemical assays), exploiting chemiluminescence as principle detection and targeting viral nucleic acids and antigens, have been performed on a permissive cell line following in vitro infection. Chemiluminescence, involving the emission of photons deriving from a chemical reaction, provided the localization and quantitative detection of analytes down to a few molecules within infected cells. In our experimental conditions, B19V transcriptional activity, leading to the production of NS and VP RNAs, has been detected early in the viral cycle (from 12 h post-infection, hpi) and before genome replication, starting at 24 hpi. The analysis of VP RNAs and related proteins suggested an inhibitory effect on capsid protein translation, as a post-transcriptional regulation events. Indeed, high levels of VP transcripts have been detected at early stages of infection while the proteins accumulated within cells only at 48–72 hpi.

Exposure to selenium, and particularly to its inorganic forms, has been hypothesized as a risk factor for amyotrophic lateral sclerosis (ALS), a fast progressing motor neuron disease with poorly understood etiology. However, no information is known about levels of inorganic and some organic selenium species in the central nervous system of ALS patients, and recent observations suggest that peripheral biomarkers of exposure are unable to predict these levels for several Se species including the inorganic forms. Using a hospital-referred cases-control series and advanced selenium speciation methods, we compared the chemical species of selenium in cerebrospinal fluid from thirty-eight ALS patients to those of thirty-eight reference neurological patients matched on age and gender. We found that higher concentrations of inorganic selenium in the form of selenite and of human serum albumin-bound selenium were associated with increased ALS risk (relative risks 3.9 (95% confidence interval 1.2–11.0) and 1.7 (1.0–2.9) for 0.1µg/l increase). Conversely, lower concentrations of selenoprotein P-bound selenium were associated with increased risk (relative risk 0.2 for 1µg/l increase, 95% confidence interval 0.04–0.8). The associations were stronger among cases age 50 years or older, who are postulated to have lower rates of genetic disease origin. These results suggest that excess selenite and human serum albumin bound-selenium and low levels of selenoprotein P-bound selenium in the central nervous system, which may be related, may play a role in ALS etiology.

We describe a case of chronic fatigue syndrome (CFS) associated to Parvovirus B19 infection where administration of intravenous immunoglobulins (IVIG), previously reported as effective, induced a paradoxical clinical response and increased viral replication. The indication of IVIG administration in the treatment of Parvovirus B19-associated CFS should be carefully reconsidered.

Argania spinosa (L.) Skeels is an endemic Moroccan species belonging to Sapotaceae family. In this work, lipophilic and aqueous extracts were obtained from leaves and subjected to a chemical profiling by MS and LC-MS/MS. Pentacyclic terpenoids were identified and quantified in the lipophilic fraction, while phenolic compounds (mainly belonging to flavonols and flavan-3-ols) were identified in the aqueous fraction. The antibacterial activities of fractions were evaluated in vitro against both reference Gram-positive and -negative bacterial strains and clinical isolates of methicillin-sensitive and methicillin-resistant Staphylococcus aureus (MSSA and MRSA); in addition, the compounds quantified as main components in each extract were assayed against reference strains. A relevant antibacterial activity was observed against reference MSSA and MRSA strains of S. aureus: for the lipophilic fraction, MIC50 values obtained were 177.8 and 170.6 μg/mL for the former and the latter, respectively, while for the aqueous fraction were 215.5 and 233.3 μg/mL. These inhibitory activities could be mainly ascribed to ursolic and oleanolic acids, among pentacyclic terpenoids, and to quercetin concerning phenolic compounds. A remarkable antibacterial activity was also observed against clinical isolates, thus argan leaves can be considered of interest in the chemotherapy of human infections.