Ides (Industrial design structure) and stylistic design engineering (sde) applied to the mobility of the future

Sustainable mobility means a series of services and means of transport designed to reduce traffic, improve air quality and cut energy consumption; it is essential to integrate the various transport systems and encourage the spread of electric vehicles. The European Commission proposes new targets for average CO2 emissions to accelerate the transition to low-emission vehicles. The goal is to reduce emissions by 40% by 2030, in line with the Paris agreements. It is not only a question of replacing private means of transport, but also of encouraging the development of new business models and more efficient use of public and freight transport. The present paper works on six main points: • New standards that help manufacturers to innovate and offer low-emission vehicles on the market • Solutions for sustainable mobility in public systems • Investments for the diffusion of infrastructures for alternative fuels • Revision of the combined transport directive which promotes the use of different means for freight transport • Development of long distance bus connections across Europe • Development of better and better batteries The diffusion of micro-mobility systems requires a general change of context. The city must keep up with technology and become smart, the regulations and all sharing phenomena must be adequate. In 2020, 80 billion objects connected to the world through 1200 satellites are estimated: urban mobility will be greatly influenced and will become a connecting element between the environment and those who live in it. The micro-mobility market in Europe could reach 150 billion dollars by 2030. As the possibilities for use increase, the number of startups for mobility sharing also increases. It is estimated that at the end of 2018 there were 5.2 million subscribers to at least one of the sharing services active in the area, one million more than the previous year. In particular, this project aims to propose an innovative, sustainable and ecological means of transport suitable for everyone and which can be a valid alternative for getting around the city

Curcumin-loaded graphene oxide flakes as an effective antibacterial system against methicillin-resistant staphylococcus aureus

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

Efficacia in vitro e in vivo dell’olio essenziale di Cinnamomum zeylanicum in nanoformulati e in sinergia con fluconazolo verso isolati di C. auris

Introduzione. C. auris è un patogeno emergente particolarmente attenzionato a livello mondiale in quanto resistente ai principali antifungini, viene spesso identificato dalle analisi di routine in maniera erronea, genera focolai tra pazienti ospedalizzati ed è resistente a disinfettanti di utilizzo comune (1). Tra le poche alternative ad oggi disponibili, l’olio essenziale di Cinnamomum zeylanicum (OE-CZ) potrebbe essere un ottimo strumento da impiegare nella lotta contro C. auris, grazie alle sue proprietà antifungine (2). Materiali e metodi. I ceppi clinici di C. auris sono stati ottenuti da emocolture positive e sottoposti a test di sensibilità agli antifungini e a 15 oli essenziali mediante tecnica della micro-diluizione. Per individuare la frazione attiva e migliorare il delivery, l’OE-CZ è stato sia frazionato con cromatografia liquida su colonna a bassa pressione che nano-incapsulato in particelle di policaprolattone (PCL). Di OE-CZ e dei prodotti della sua lavorazione è stata valutata sia la qualità chimica (GC-MS accoppiato allo spazio di testa) che l’efficacia antifungina unitamente alle principali componenti delle frazioni. La tossicità e l’efficacia di OE-CZ libero (L-OE-CZ) e nano-incapsulato (N-OE-CZ) sono state testate in vivo su G. mellonella. Il metodo di titolazione a scacchiera è stato usato per valutare la sinergia tra l’L-OE-CZ o N-OE-CZ e gli antimicotici (fluconazolo e micafungina). Il meccanismo d’azione è stato indagato valutando l’attività ATPasica in cellule di C.auris mediante kit ELISA, e la quantità di fluconazolo intracellulare con UPLC-MS/MS. Risultati. Tra i diversi OEs testati, l’OE-CZ era il più attivo (MIC90=0,06% v/v; MFC90=0,06% v/v). Tutti i ceppi di C. auris mostravano resistenza al fluconazolo (MIC>1024 μg/ml). La cinnamaldeide è il maggior costituente di L-OE-CZ (66,1 %). L’analisi dello spazio di testa indica che la quantità di quest’ultima aumentava da L-OE-CZ (24.2%) a di N-OE-CZ (80,1%) con una conseguente migliore efficacia (MIC90 ed MFC90=0.02 % v/v). Solo L-OE-CZ mostrava sinergia con il fluconazolo (FIC = 0,26 ± 0,14) e con la micafungina (FIC= 0,23 ± 0,13 e FIC=0,32 ± 0,13). In vivo, nessuna concentrazione di L-OE-CZ testata è risultata tossica e l’efficacia migliore era data dalla sinergia individuata in vitro (fluconazolo 2,5 μg/mL + OE 0,002% v/v) con parallela diminuzione dell’attività ATPasica (p<0,0001) e aumento della quantità di fluconazolo (p<0,002) intracellulare rispetto al controlli non trattati. Conclusioni e discussione. L’analisi di qualità dell’OE di C. zeylanicum individua la cinnamaldeide (66.1%) e il cinnamil-acetato (5.5%) come principali componenti. La nanoincapsulazione dell’OE comporta l’incorporazione selettiva di specifici principi attivi, come cinnamaldeide ed eugenolo, con il conseguente incremento delle proprietà antifungine del formulato. Dai test di sinergia emerge che solo L-OE-CZ va in sinergia con il fluconazolo e con la micafungina, mentre N-OE-CZ perde questa proprietà: ciò è probabilmente dovuto alla mancanza di componenti importanti minoritari del fitocomplesso che vengono persi nel processo di incapsulazione. I dati in vivo indicano sia la sicurezza d’uso delle concentrazioni testate che l’efficacia della sinergia tra OE e fluconazolo. Sebbene non sia ancora noto il meccanismo con cui L-OE-CZ agisca sul meccanismo di resistenza della cellula è possibile presupporre che questo coinvolga l’inibizione delle pompe di efflusso attivo ATP dipendenti permettendo al farmaco di permanere nella cellula e agire come farebbe in cellule di Candida spp. sensibil

The polyamine N-acetyltransferase-like enzyme PmvE plays a role in the virulence of Enterococcus faecalis

We previously showed that the mutant strain of Enterococcus faecalis lacking the transcriptional regulator SlyA is more virulent than the parental strain. We hypothesized that this phenotype was due to overexpression of the second gene of the slyA operon, ef_3001, renamed pmvE (for polyamine metabolism and virulence of E. faecalis). PmvE shares strong homologies with N(1)-spermidine/spermine acetyltransferase enzymes involved in the metabolism of polyamines. In this study, we used an E. faecalis strain carrying the recombinant plasmid pMSP3535-pmvE (V19/p3535-pmvE), which allows the induction of pmvE by addition of nisin. Thereby, we showed that the overexpression of PmvE increased the virulence of E. faecalis in the Galleria mellonella infection model, as well as the persistence within peritoneal macrophages. We were also able to show a direct interaction between the His-tagged recombinant PmvE (rPmvE) protein and putrescine by the surface plasmon resonance (SPR) technique on a Biacore instrument. Moreover, biochemical assays showed that PmvE possesses an N-acetyltransferase activity toward polyamine substrates. Our results suggest that PmvE contributes to the virulence of E. faecalis, likely through its involvement in the polyamine metabolism

The Polyamine N-Acetyltransferase-Like Enzyme PmvE Plays a Role in the Virulence of Enterococcus faecalis

International audienceWe previously showed that the mutant strain of Enterococcus faecalis lacking the transcriptional regulator SlyA is more virulent than the parental strain. We hypothesized that this phenotype was due to overexpression of the second gene of the slyA operon, ef_3001, renamed pmvE (for polyamine metabolism and virulence of E. faecalis). PmvE shares strong homologies with N 1-spermidine/spermine acetyltransferase enzymes involved in the metabolism of polyamines. In this study, we used an E. faecalis strain carrying the recombinant plasmid pMSP3535-pmvE (V19/p3535-pmvE), which allows the induction of pmvE by addition of nisin. Thereby, we showed that the overexpression of PmvE increased the virulence of E. faecalis in the Galleria mellonella infection model, as well as the persistence within peritoneal macrophages. We were also able to show a direct interaction between the His-tagged recombinant PmvE (rPmvE) protein and putrescine by the surface plasmon resonance (SPR) technique on a Biacore instrument. Moreover, biochemical assays showed that PmvE possesses an N-acetyltransferase activity toward polyamine substrates. Our results suggest that PmvE contributes to the virulence of E. faecalis, likely through its involvement in the polyamine metabolism

Inactivation of the Response Regulator AgrA Has a Pleiotropic Effect on Biofilm Formation, Pathogenesis and Stress Response in Staphylococcus lugdunensis

Staphylococcus lugdunensis is a coagulase-negative Staphylococcus that emerges as an important opportunistic pathogen. However, little is known about the regulation underlying the transition from commensal to virulent state. Based on knowledge of S. aureus virulence, we suspected that the agr quorum sensing system may be an important determinant for the pathogenicity of S. lugdunensis. We investigated the functions of the transcriptional regulator AgrA using the agrA deletion mutant. AgrA played a role in cell pigmentation: DargA mutant colonies were white while the parental strains were slightly yellow. Compared with the wild-type strain, the DargA mutant was affected in its ability to form biofilm and was less able to survive in mice macrophages. Moreover, the growth of DagrA was significantly reduced by the addition of 10% NaCl or 0.4 mM H2O2 and its survival after 2 h in the presence of 1 mM H2O2 was more than 10-fold reduced. To explore the mechanisms involved beyond these phenotypes, the DagrA proteome and transcriptome were characterized by mass spectrometry and RNA-Seq. We found that AgrA controlled several virulence factors as well as stress-response factors, which are well correlated with the reduced resistance of the DagrA mutant to osmotic and oxidative stresses. These results were not the consequence of the deregulation of RNAIII of the agr system, since no phenotype or alteration of the proteomic profile has been observed for the DRNAIII mutant. Altogether, our results highlighted that the AgrA regulator of S. lugdunensis played a key role in its ability to become pathogenic

Re-evaluating positive serum samples for SARS-CoV-2-specific IgA and IgG antibodies using an in-house serological assay

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Phytocomplex influences antimicrobial and health properties of concentrated glycerine macerates

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