Virginia Fuochi

Evaluation of resistance to low pH and bile salts of human Lactobacillus spp. isolates

There are nearly 100 trillion bacteria in the intestine that together form the intestinal microbiota. They are ‘good’ bacteria because they help to maintain a physiological balance and are called probiotics. Probiotics must have some important characteristics: be safe for humans, be resistant to the low pH in the stomach, as well as bile salts and pancreatic juice. Indeed, their survival is the most important factor, so that they can arrive alive in the intestine and are able to form colonies, at least temporarily. The aim of our study was the evaluation of resistance of Lactobacillus isolates from fecal and oral swabs compared to that found in a commercial product. Seven strains were randomly chosen: L. jensenii, L. gasseri, L. salivarius, L. fermentum, L. rhamnosus, L. crispatus, and L. delbrueckii. We observed a large variability in the results: L. gasseri and L. fermentum were the most resistance to low pH, while only L. gasseri showed the best survival rate to bile salts. Interestingly, the commercial product did not show tolerance to both low pH and bile salts.

Probiotic Properties of Lactobacillus fermentum Strains Isolated from Human Oral Samples and Description of their Antibacterial Activity

BACKGROUND Gram positive bacteria produce peptides, defined bacteriocins which exhibit good antibacterial activity. OBJECTIVE We evaluated the ability of L. fermentum to produce bacteriocins having therefore, good probiotic features and finally, be safe towards microglial cells. METHOD Eight wild strains, identified using molecular techniques, were investigated for the evaluation of resistance to bile salts, low pH, H2O2 production, biofilm formation, antibacterial activity and safety on microglia cells (BV2). RESULTS The determination of the susceptibility/resistance profile showed that the strains are sensitive to the antibiotics tested. All strains showed a good tolerability to extremely low pH as well as resisting in presence of bile salts. In addition, the strains showed excellent activity against pathogens and one of them (LAC 42) showed activity also against Pseudomonas aeruginosa and Klebsiella pneumoniae. Finally, LAC 42 and its active compound did not change microglia cell viability following 24h exposure. Our data on this antibacterial molecule suggest that it is a compound with low molecular weight and with highly hydrophilic component. CONCLUSION These results describe the characteristics of Lactobacillus strains and provide evidences for their possible use as new potential probiotic. In addition, other studies are now warranted to exploit the antibacterial activity of the supernatant LAC 42 and for its complete chemical characterization.

A Method for Efficient Loading of Ciprofloxacin Hydrochloride in Cationic Solid Lipid Nanoparticles: Formulation and Microbiological Evaluation

The aim of the study was the production of solid lipid nanoparticles (SLN) loaded with ciprofloxacin (CIP) through two different production techniques, quasi-emulsion solvent diffusion (QESD) and solvent injection (SI). In order to efficaciously entrap the commercial salt form (hydrochloride) of the antibiotic in these lipid systems, a conversion of CIP hydrochloride to the free base was realized in situ, through the addition of triethylamine. To ensure physical stability to the carriers over time and ameliorate the interaction with bacterial cell membranes, positively charged SLN were produced by addition of the cationic lipid didecyldimethylammonium bromide (DDAB). Homogeneous SLN populations with a mean particle sizes of 250–350 nm were produced by both methods; drug encapsulation was over 85% for most samples. The SLN were physically stable for up to nine months both at 4 °C and 25 °C, although the former condition appears more suitable to guarantee the maintenance of the initial particle size distribution. As expected, CIP encapsulation efficiency underwent a slight reduction after nine months of storage, although the initial high drug content values would ensure a residual concentration of the antibiotic in the SLN still appropriate to exert an acceptable antibacterial activity. Selected SLN formulations were subjected to an in vitro microbiological assay against different bacterial strains, to verify the effect of nanoencapsulation on the cell growth inhibitory activity of CIP. In general, CIP-SLN produced without DDAB showed MIC values for CIP comparable to those of the free drug. Conversely, addition of increasing percentages of the cationic lipid, reflected by a progressive increase of the positive value of the Zeta potential, showed a variety of MIC values against the various bacterial strains, but with values 2–4 order of dilution lower than free CIP. An hypothesis of the effect of the cationic lipid upon the increased antibacterial activity of CIP in the nanocarriers is also formulated.

Antioxidant and antimicrobial properties of Casteanea sativa Miller chestnut honey produced on Mount Etna (Sicily)

Abstract The aim of this study was the evaluation of antibacterial and antioxidant properties of Monofloral Etna Castanea sativa Miller honeys. Escherichia coli ATCC 25,922, Pseudomonas aeruginosa ATCC 27,853, Enterococcus faecalis ATCC 29,211 and Staphylococcus aureus ATCC 29,213 were investigated for their susceptibilities to two different honeys. Antioxidant activity was evaluated by ORAC, NO scavenger assays, FRAP and DPPH. Antioxidant activity and antibacterial properties were compared with chestnut honeys from different geographical areas and with Manuka honey. UPLC-MS/MS was used for major components characteri sation.

Antimicrobial and Anti-Proliferative Effects of Skin Mucus Derived from Dasyatis pastinaca (Linnaeus, 1758)

Resistance to chemotherapy occurs in various diseases (i.e., cancer and infection), and for this reason, both are very difficult to treat. Therefore, novel antimicrobial and chemotherapic drugs are needed for effective antibiotic therapy. The aim of the present study was to assess the antimicrobial and anti-proliferative effects of skin mucus derived from Dasyatis pastinaca (Linnaeus, 1758). Our results showed that skin mucus exhibited a significant and specific antibacterial activity against Gram-negative bacteria but not against Gram-positive bacteria. Furthermore, we also observed a significant antifungal activity against some strains of Candida spp. Concerning anti-proliferative activity, we showed that fish mucus was specifically toxic for acute leukemia cells (HL60) with an inhibition of proliferation in a dose dependent manner (about 52% at 1000 μg/mL of fish skin mucous, FSM). Moreover, we did not observe effects in healthy cells, in neuroblastoma cells (SH-SY5Y), and multiple myeloma cell lines (MM1, U266). Finally, it exhibited strong expression and activity of chitinase which may be responsible, at least in part, for the aforementioned results.

Key Roles of Human Polymorphonuclear Cells and Ciprofloxacin in Lactobacillus Species Infection Control

ABSTRACT Lactobacilli have the potential to act as reservoirs of antibiotic resistance genes similar to those found in human pathogens, with the risk of transferring these genes to many pathogenic bacteria. In this study, we investigated the role of human polymorphonuclear cells (PMNs) against Lactobacillus spp. both resistant and susceptible to ciprofloxacin (a fluoroquinolone) and the effect of ciprofloxacin on the interaction between PMNs and three Lactobacillus spp. with different patterns of susceptibility to this drug. Hence, the primary functions of PMNs, such as phagocytosis and bacterial intracellular killing, against lactobacilli were investigated. The rate of PMN phagocytosis was high for ciprofloxacin-sensitive and ciprofloxacin-resistant strains. The patterns of intracellular killing of ciprofloxacin-sensitive and ciprofloxacin-resistant strains by PMNs underline that PMNs alone were able to kill lactobacilli. The addition of ciprofloxacin to PMNs did not result in a significant increase in the bacterial uptake by phagocytes. On the contrary, ciprofloxacin had a marked effect on PMN intracellular killing, resulting in increased numbers of killed ciprofloxacin-sensitive bacteria in comparison with antibiotic-free controls. Our data show that by itself, the profile of antibiotic resistance does not constitute an intrinsic factor of greater or lesser pathogenicity toward the host. The ability of PMNs to kill a diverse array of bacterial pathogens is essential for human innate host defense, primarily in immunocompromised patients.

Mepolizumab in the management of severe eosinophilic asthma in adults: current evidence and practical experience

Asthma is a chronic inflammatory condition involving the airways with varying pathophysiological mechanisms, clinical symptoms and outcomes, generally controlled by conventional therapies including inhaled corticosteroids and long-acting β2 agonists. However, these therapies are unable to successfully control symptoms in about 5–10% of severe asthma patients. Atopic asthma, characterized by high immunoglobulin (Ig)E or eosinophilia, represents about 50% of asthmatic patients. Interleukin (IL)-5 is the main cytokine responsible of activation of eosinophils, hence therapeutic strategies have been investigated and developed for clinical use. Biologics targeting IL-5 and its receptor (first mepolizumab and subsequently, reslizumab and benralizumab), have been recently approved and used as add-on therapy for severe eosinophilic asthma resulting in a reduction in the circulating eosinophil count, improvement in lung function and exacerbation reduction in asthma patients. Despite these biologics having been approved for stratified severe asthma patients that remain uncontrolled with high doses of conventional therapy, a number of patients may be eligible for more than one biologic. Presently, the lack of head-to-head studies comparing the biological agents among themselves and with conventional therapy make the choice of optimal therapy for each patient a challenge for clinicians. Moreover, discontinuation of these treatments, implications for efficacy or adverse events, in particular in long-term treatment, and needs for useful biomarkers are still matters of debate. In this review we evaluate to date, the evidence on mepolizumab that seems to demonstrate it is a well-tolerated and efficacious regimen for use in severe eosinophilic asthma, though more studies are still required.

Commentary: Inflammatory and Oxidative Responses Induced by Exposure to Commonly Used e-Cigarette Flavoring Chemicals and Flavored e-Liquids without Nicotine

Citation: Caruso M, Li Volti G, Furneri PM, Fuochi V, Emma R and Polosa R (2018) Commentary: Inflammatory and Oxidative Responses Induced by Exposure to Commonly Used e-Cigarette Flavoring Chemicals and Flavored e-Liquids without Nicotine. Front. Physiol. 9:1240. doi: 10.3389/fphys.2018.01240 Commentary: Inflammatory and Oxidative Responses Induced by Exposure to Commonly Used e-Cigarette Flavoring Chemicals and Flavored e-Liquids without Nicotine

Electronic cigarette vapour enhances pneumococcal adherence to airway epithelial cells under abnormal conditions of exposure

We read with great interest the research article by Miyashita et al. [1], in which the authors showed that e-cigarette aerosol emissions increase platelet-activating factor receptor (PAFR) expression and, consequently, Streptococcus pneumoniae adhesion to human airway epithelial cells. These findings led authors to conclude that e-cigarette use has the potential to increase susceptibility to pneumococcal infection. Unfortunately, the evidence presented in the paper is inadequate to provide much confidence in this conclusion. E-cigarette vapour enhances pneumococcal adherence to airway epithelial cells under “abnormal” conditions of exposure http://ow.ly/O9J030keaYe

Nanosized devices as antibiotics and antifungals delivery: past, news, and outlook

Abstract Clinical treatment of bacterial and fungal infections often meets many drawbacks associated with the low bioavailability of active agents, adverse side effects, difficulty in penetrating and concentrating inside the target cells, intrinsic or acquired resistance of microorganisms. In recent years, nanotechnology has driven efforts toward this therapeutic area, proposing efficient colloidal (nanosized) delivery systems (NDDS) that have attained in some cases a market significance and clinical relevance. NDDS can control the release of loaded drugs or target their distribution within the body, focusing selectively their activity in the infected cells, thus reducing side effects. Loading antibacterials and antifungals in nanocarriers through physical encapsulation, adsorption, or chemical conjugation, makes it possible to overwhelm their poor solubility and improve pharmacokinetics and therapeutic index, compared to the neat drugs. Among the most promising NDDS are polymer- and lipid-based nanocarriers, such as liposomes, polymeric nanoparticles and nanocapsules, micelles, and lipid nanoparticles. In this chapter recent developments in the field of NDDS encapsulating antibacterial and antifungal drugs are discussed. Specific attention has been given to the pharmaceutical, microbiological, and clinical outcomes of systems for which the authors have provided in vitro microbiological data and, more hopefully, in vivo results.