Federico Pepi

Effects of Mentha suaveolens Essential Oil Alone or in Combination with Other Drugs in Candida albicans

Candidosis is the most important cause of fungal infections in humans. The yeast Candida albicans can form biofilms, and it is known that microbial biofilms play an important role in human diseases and are very difficult to treat. The prolonged treatment with drugs has often resulted in failure and resistance. Due to the emergence of multidrug resistance, alternatives to conventional antimicrobial therapy are needed. This study aims to analyse the effects induced by essential oil of Mentha suaveolens Ehrh (EOMS) on Candida albicans and its potential synergism when used in combination with conventional drugs. Morphological differences between control and EOMS treated yeast cells or biofilms were observed by scanning electron microscopy and transmission electron microscopy (SEM and TEM resp.,). In order to reveal the presence of cell cycle alterations, flow cytometry analysis was carried out as well. The synergic action of EOMS was studied with the checkerboard method, and the cellular damage induced by different treatments was analysed by TEM. The results obtained have demonstrated both the effects of EOMS on C. albicans yeast cells and biofilms and the synergism of EOMS when used in combination with conventional antifungal drugs as fluconazole (FLC) and micafungin (MCFG), and therefore we can hypothesize on its potential use in therapy. Further studies are necessary to know its mechanism of action.

Electron‐Transfer Kinetics of Microperoxidase‐11 Covalently Immobilised onto the Surface of Multi‐Walled Carbon Nanotubes by Reactive Landing of Mass‐Selected Ions

Controlled deposition of biological molecules on nanostructured materials is a basic step towards the realisation of biochip components. In this study we report the investigation of the first covalent immobilisation of mass-selected redox protein on a carboxyl-functionalised multi-walled carbon nanotube (MWCNT) electrode surface by means of ion soft landing. The immobilised protein maintains its biochemical properties, displaying an excellent electrochemical behaviour on the electrode surface. The deposition of mass-selected ions is influenced by several factors, including the charge state and the collision energy of the projectile ions. To elucidate the mechanism involved in the protein reactive landing onto the MWCNT surface, the data obtained from cyclic voltammetry experiments were modelled according to the Marcus theory. The proposed method opens up the way to the development of a new generation of biocomponents with potential use in biosensors, diagnostics, biofuel cells and bioactive films.

Ionic Fluorination of Carbon Monoxide as a Route to Gasphase Carbonylation of Inert CH and NH Bonds

Gaseous FCO+ ions from the ionization of mixtures of nitrogen trifluoride and carbon monoxide execute selective and efficient CO-functionalization of the C-H bonds of benzene and toluene and of the N-H bond of ammonia. The occurrence of these carbonylation reactions has been unambiguously ascertained by Fourier-transform ion cyclotron resonance (FT-ICR) spectrometry, and the details of the structure and the mechanism of formation of the precursor FCO+ ions have been investigated. FT-ICR experiments show that these ions, structurally assigned as F-C-O+ by collisionally activated dissociation (CAD) spectrometry, arise from the reaction of CO.+ with NF3 and of NF+2 with CO. Combining the latter F+ transfer with the independently observed fluoride-ion abstraction by FCO+ from NF3 results in a catalytic cycle in which gaseous NF+2 ions promote the conversion of carbon monoxide into carbonic difluoride, F2 CO, with nitrogen trifluoride as the source of F.

Soft-landed protein voltammetry: a tool for redox protein characterization.

Microperoxidase-11 (MP-11) was first soft landed onto the gold surface of a screen-printed electrode. Intact protein deposition was verified by time-of-flight secondary ion mass spectrometry. The coupling of soft landing with electrochemical techniques allowed unique information to be obtained about the deposition features. A full characterization of the direct electron-transfer properties was performed by modeling data obtained from cyclic voltammetry experiments; calculated values of kinetic electron-transfer constant, formal redox potential, and reorganization energy allow us to hypothesize the mechanism involved in soft landing immobilization and demonstrate the different conformation of the enzyme deposited from two different charged species. The strong interaction between MP-11 and the gold surface and long-term stability of the functionalized electrode characterizes the peculiar features of this procedure, which enhance its power with respect to the existing immobilization procedure and ensure its suitability for those practical applications that could benefit from an unmediated bridgeless bioeletrochemical electron transfer (e.g., biosensor transducers or electrode elements in biofuel cells).

In vitro inhibition of herpes simplex virus type 1 replication by Mentha suaveolens essential oil and its main component piperitenone oxide

Several essential oils exert in vitro activity against bacteria and viruses and, among these latter, herpes simplex virus type 1 (HSV-1) is known to develop resistance to commonly used antiviral agents. Thus, the effects of the essential oil derived from Mentha suaveolens (EOMS) and its active principle piperitenone oxide (PEO) were tested in in vitro experimental model of infection with HSV-1. The 50% inhibitory concentration (IC50) was determined at 5.1μg/ml and 1.4μg/ml for EOMS and PEO, respectively. Australian tea tree oil (TTO) was used as control, revealing an IC50 of 13.2μg/ml. Moreover, a synergistic action against HSV-1 was observed when each oil was added in combination with acyclovir. In order to find out the mechanism of action, EOMS, PEO and TTO were added to the cells at different times during the virus life-cycle. Results obtained by yield reduction assay indicated that the antiviral activity of both compounds was principally due to an effect after viral adsorption. Indeed, no reduction of virus yield was observed when cells were treated during viral adsorption or pre-treated before viral infection. In particular, PEO exerted a strong inhibitory effect by interfering with a late step of HSV-1 life-cycle. HSV-1 infection is known to induce a pro-oxidative state with depletion of the main intracellular antioxidant glutathione and this redox change in the cell is important for viral replication. Interestingly, the treatment with PEO corrected this deficit, thus suggesting that the compound could interfere with some redox-sensitive cellular pathways exploited for viral replication. Overall our data suggest that both EOMS and PEO could be considered good candidates for novel anti-HSV-1 strategies, and need further exploration to better characterize the targets underlying their inhibition.

Essential Oil Extraction, Chemical Analysis and Anti-Candida Activity of Calamintha nepeta (L.) Savi subsp. glandulosa (Req.) Ball-New Approaches.

A comprehensive study on essential oils extracted from different Calamintha nepeta (L.) Savi subsp. glandulosa (Req.) Ball samples from Tarquinia (Italy) is reported. In this study, the 24-h steam distillation procedure for essential oil preparation, in terms of different harvesting and extraction times, was applied. The Gas chromatography–mass spectrometry (GC/MS) analysis showed that C. nepeta (L.) Savi subsp. glandulosa (Req.) Ball essential oils from Tarquinia belong to the pulegone-rich chemotype. The analysis of 44 samples revealed that along with pulegone, some other chemicals may participate in exerting the related antifungal activity. The results indicated that for higher activity, the essential oils should be produced with at least a 6-h steam distillation process. Even though it is not so dependent on the period of harvesting, it could be recommended not to harvest the plant in the fruiting stage, since no significant antifungal effect was shown. The maximum essential oil yield was obtained in August, with the highest pulegone percentage. To obtain the oil with a higher content of menthone, September and October should be considered as the optimal periods. Regarding the extraction duration, vegetative stage material gives the oil in the first 3 h, while material from the reproductive phase should be extracted at least at 6 or even 12 h.

Gas-phase reactions of nitronium ions with acetylene and ethylene: an experimental and theoretical study

A comparative study of the gas-phase reactions of NO2+ with acetylene and ethylene was performed by using FT-ICR, MIKE, CAD, and NfR/ CA mass spectrometric techniques, in conjunction with ab initio calculations at the MP2/6-31+G* level of theory. Both reactions proceed according to the same mechanism, that is, 1,3-dipolar cycloaddition, but yield products of different stability. The C2H2NO2+ adduct from acetylene has an aromatic character and hence is highly stabilized with respect to the C2H4NO2+ adduct from ethylene. Both cycloadducts tend to isomerize into O-nitroso derivatives, that is, nitrosated ketene and nitrosated acetaldehyde, which represent the thermodynamically most stable products from the addition of NO2+ to acetylene and ethylene, respectively. As prototypal examples of the reactivity of free nitronium ions with most simple pi systems, the reactions investigated are useful starting points to model the mechanism of aromatic nitration.

Multidisciplinary Approach to Determine the Optimal Time and Period for Extracting the Essential Oil from Mentha suaveolens Ehrh

A comprehensive study on essential oils (EOs) extracted from some Mentha suaveolens L. samples, collected in the countryside of Tarquinia, is reported. In this study, the procedure for essential oil preparation, in terms of harvesting and extraction time, was analyzed in detail for the first time. The GC/MS analysis, carried out on 18 samples, revealed that piperitenone oxide (PO), the main essential oils’ chemical constituent, is primarily responsible for the related antifungal activity. Nevertheless, EOs with lower PO content indicate that other chemicals, such as para-cymenene, may participate in exerting the EOs’ antifungal effect. Furthermore, the bacterial reverse mutation assay highlighted lack of mutagenic effect in all tested samples. Analysis of the results indicated that for higher activity, the essential oils should be produced with 3 h maximum hydrodistillation, regardless of the harvesting time. Differently, the maximum essential oil yield can be obtained in August and the highest piperitenone oxide percentage is obtainable in July.

Gellan gum and polyethylene glycol dimethacrylate double network hydrogels with improved mechanical properties

Combining different polymeric systems can be a useful tool to create new networks with different characteristics with respect to the starting materials. In this work, hydrogels composed of gellan gum (GG) and polyethylene glycol dimethacrylate (PEG-DMA) were realized to overcome the fragility problems of physical gels of GG, which limit their biological application as scaffold for tissue engineering. The two polymeric systems were combined using different synthetic approaches, with particular attention to the double network strategy (DN). The influence of several parameters on the mechanical properties, such as the time of diffusion and the molecular weight of PEG-DMA, were evaluated by rheological studies and compressive texture analyses. The hydrogels were also investigated for their ability to swell and release model molecules with different sterical hindrances, such as vitamin B12 and myoglobin. Finally, to estimate the biological safety of the hydrogels, their effect on mitochondrial function of human fibroblasts was investigated.

Direct Experimental Evidence for the H2O+O2− Charge Transfer Complex: Crucial Support to Atmospheric Photonucleation Theory

Where does rain come from? The first experimental evidence for the existence of the (H(2)O(+)O(2)(-)) charge transfer complex, predicted to play a key role as a nucleation center for atmospheric vapor photonucleation (see picture), is provided by the powerful neutralization - reionization mass spectrometric technique.