Serratiopeptidase reduces the invasion of osteoblasts by Staphylococcus aureus

Finding new strategies to counteract periprosthetic infection and implant failure is a main target in orthopedics. Staphylococcus aureus, the leading etiologic agent of orthopedic implant infections, is able to enter and kill osteoblasts, to stimulate pro-inflammatory chemokine secretion, to recruit osteoclasts, and to cause inflammatory osteolysis. Moreover, by entering eukaryotic cells, staphylococci hide from the host immune defenses and shelter from the extracellular antibiotics. Thus, infection persists, inflammation thrives, and a highly destructive osteomyelitis occurs around the implant. The ability of serratiopeptidase (SPEP), a metalloprotease by Serratia marcescens, to control S. aureus invasion of osteoblastic MG-63 cells and pro-inflammatory chemokine MCP-1 secretion was evaluated. Human osteoblast cells were infected with staphylococcal strains in the presence and in the absence of SPEP. Cell proliferation and cell viability were also evaluated. The release of pro-inflammatory chemokine MCP-1 was evaluated after the exposure of the osteoblast cells to staphylococcal strains. The significance of the differences in the results of each test and the relative control values was determined with Student’s t-test. SPEP impairs their invasiveness into osteoblasts, without affecting the viability and proliferation of bone cells, and tones down their production of MCP-1. We recognize SPEP as a potential tool against S. aureus bone infection and destruction

Education in Soil Science: the Italian approach

The Italian Society of Soil Science (SISS) was founded in Florence on February 18th, 1952. It is an association legally acknowledged by Decree of the President of the Italian Republic in February 1957. The Society is member of the International Union of Soil Sciences (IUSS) of the European Confederation of Soil Science Societies (ECSSS) and collaborates with several companies, institutions and organizations having similar objectives or policy aspects. SISS promotes progress, coordination and dissemination of soil science and its applications encouraging relationships and collaborations among soil lovers. Within the SISS there are Working Groups and Technical Committees for specific issues of interest. In particular: \u2022 the Working Group on Pedotechniques; \u2022 the Working Group on Hydromorphic and Subaqueous Soils and \u2022 the Technical Committee for Soil Education and Public Awareness. In this communication we wish to stress the activities developed since its foundation by SISS to spread soil awareness and education in Italy through this last Technical Committee, focusing also the aspect concerning grants for young graduates and PhD graduates to stimulate the involvement of young people in the field of soil science

Endothelial cells, endoplasmic reticulum stress and oxysterols

Oxysterols are bioactive lipids that act as regulators of lipid metabolism, inflammation, cell viability and are involved in several diseases, including atherosclerosis. Mounting evidence linked the atherosclerosis to endothelium dysfunction; in fact, the endothelium regulates the vascular system with roles in processes such as hemostasis, cell cholesterol, hormone trafficking, signal transduction and inflammation. Several papers shed light the ability of oxysterols to induce apoptosis in different cell lines including endothelial cells. Apoptotic endothelial cell and endothelial denudation may constitute a critical step in the transition to plaque erosion and vessel thrombosis, so preventing the endothelial damaged has garnered considerable attention as a novel means of treating atherosclerosis. Endoplasmic reticulum (ER) is the site where the proteins are synthetized and folded and is necessary for most cellular activity; perturbations of ER homeostasis leads to a condition known as endoplasmic reticulum stress. This condition evokes the unfolded protein response (UPR) an adaptive pathway that aims to restore ER homeostasis. Mounting evidence suggests that chronic activation of UPR leads to cell dysfunction and death and recently has been implicated in pathogenesis of endothelial dysfunction. Autophagy is an essential catabolic mechanism that delivers misfolded proteins and damaged organelles to the lysosome for degradation, maintaining basal levels of autophagic activity it is critical for cell survival. Several evidence suggests that persistent ER stress often results in stimulation of autophagic activities, likely as a compensatory mechanism to relieve ER stress and consequently cell death. In this review, we summarize evidence for the effect of oxysterols on endothelial cells, especially focusing on oxysterols-mediated induction of endoplasmic reticulum stress

Spatial microbial community structure and biodiversity analysis in "extreme" hypersaline soils of a semiarid Mediterranean area

In recent years specific attention has been paid on the biotechnological potential of microorganisms in extreme soils, in particular in saline soils. Salinity is one of the most widespread soil degradation processes on the Earth, and saline soils can be defined as extreme soils or border line habitats in which several factors, as high salt content, may limit the growth of organisms. In this study, the physical, chemical and microbiological soil properties were investigated in the shallower horizon of natural salt-affected soils in Sicily (Italy). The main aim of the research was to evaluate the structure and diversity of bacterial and archaeal communities by terminal-restriction fragment length polymorphism (T-RFLP) according to arbitrary different classes of vegetation and salt crust cover in soils. Furthermore, the structure of microbial communities was assessed considering the heterogeneity of physical-chemical properties of the habitat under investigation, as a function of vegetation, crust cover, and salinity classes. The results provided information on the type of distribution of different microbial community composition and diversity as a function of both vegetation and crust cover as well as salinity classes. In particular, the archaeal community showed a richness and diversity significantly affected by the spatial gradients of soil salinity, conversely, the bacterial one showed a decreasing trend with increasing gradient of soil salinity. The T-RFLP cluster analysis showed the formation of two groups for both bacterial and archaeal community, significantly (. p<. 0.05) influenced by sand and silt content, electrical conductivity (EC. e), vegetation cover percentage, salt crust and for by texture composition. In particular, the discriminant analysis obtained for the different salt crust classes for archaeal community stressed the membership of one of the two clusters to the class with the lower salt crust percentage (0-40%)

Comparison of the action of different proteases on virulence properties related to the staphylococcal surface.

AIMS: The purpose of this study was to evaluate the antimicrobial efficacy of five different proteases belonging to two different families on Staphylococcus aureus and Staphylococcus epidermidis strains. METHODS AND RESULTS: We used three serine proteases and two metalloproteases in single species biofilm formation assays and in human cell invasion processes. Following each protease incubation with bacterial cells, surface protein patterns were analysed by SDS-PAGE and zymography. Some differently expressed proteins were identified by mass spectrometry. CONCLUSIONS: The effect of tested proteases on biofilm formation was not related to the protease category but was strain-dependent and was related to the biofilm formation capacity of each staphylococcal strain. Some proteases showed a nonspecific and indiscriminate effect on surface proteins, while others induced a discrete and reproducible action on protein profiles. SIGNIFICANCE AND IMPACT OF THE STUDY: The inhibition of the surface-related virulence factors is a promising avenue to overcome persistent infections caused by bacterial biofilms. To this end, we show here that proteases, in particular the metalloprotease serratiopeptidase, can interfere with adhesion and invasion of eukaryotic cells and biofilm formation in staphylococci and their use could represent a viable treatment for the development of novel combination therapie

Distribution patterns of fungi and bacteria in saline soils

Saline soils are environments characterized by uneven temporal and spatial water distribution and localized high concentrations of salts. Spatial distribution patterns of fungi and bacteria in saline soils, and the link between microbial community dynamics and salts accumulation are critical issues throughout the world (Ettema, Wardle 2002). This study was focused on spatial distribution patterns of soil fungi and bacteria in a saline soil located in Piana del Signore (Gela, Italy) where some ecological variables acted as shaping factors in aboveground and belowground communities distribution. Bacterial, archaeal, and fungal communities diversity and distribution in ten soil sites (A horizons, 0-10cm), were characterized by 16S rDNA genes with T-RFLP method. Pyrosequencing-based analysis of the V2-V3 16S rRNA gene region was performed to characterize the sites on the basis of bacterial groups distribution, diversity and assemblage. To better investigate the ecological niches of some of the main culturable species of this environment, it was carried out the isolation and identification of the fungal flora from soil, using Warcup plating within two different salt concentrations (NaCl 5% and 15%), combined with a metabolic screening of some representative isolates (Di Lonardo et al., 2013). A natural gradient of soil salinity shaped the distribution of microbial species in the environment. The different concentration of salt (NaCl), and calcium sulfate (Ca2SO4) in soil influenced the structure and distribution of the microbial communities even when comparing neighboring areas within a 50 m scale. Some bacterial phyla, together with some fungal species, appeared spread in the whole area, independently of the salinity gradient, thus highlighting the presence of organisms with a very different survival strategy in such an extreme environment. In conclusion, the organization and diversity of microbial taxa at a spatial scale reflected the scales of heterogeneity of physical and chemical properties of the habitat under investigation

Salinity and Bacterial Diversity: To What Extent Does the Concentration of Salt Affect the Bacterial Community in a Saline Soil?

In this study, the evaluation of soil characteristics was coupled with a pyrosequencing analysis of the V2-V3 16S rRNA gene region in order to investigate the bacterial community structure and diversity in the A horizon of a natural saline soil located in Sicily (Italy). The main aim of the research was to assess the organisation and diversity of microbial taxa using a spatial scale that revealed physical and chemical heterogeneity of the habitat under investigation. The results provided information on the type of distribution of different bacterial groups as a function of spatial gradients of soil salinity and pH. The analysis of bacterial 16S rRNA showed differences in bacterial composition and diversity due to a variable salt oncentration in the soil. The bacterial community showed a statistically significant spatial variability. Some bacterial phyla appeared spread in the whole area, whatever the salinity gradient. It emerged therefore that a patchy saline soil can not contain just a single microbial community selected to withstand extreme osmotic phenomena, but many communities that can be variously correlated to one or more environmental parameters. Sequences have been deposited to the SRA database and can be accessed on ID Project PRJNA241061

In situ remediation of polluted Spolic Technosols using Ca(OH)2 and smectitic marlstone

Technosols are soils developed on non-traditional substrates and containing large quantities of materials mostly due to intensive human industrial activity, such as artefacts. The increasing number of sites affected by Technosols and their impact on the environment as growing media for plants or as source of pollutants require an understanding of their functioning and evolution, above all the knowledge on the transport of toxic substances from contaminated technogenic soils to groundwater. A case study on properties, remediation and evaluation of Technosols made up by vitrified fly ash and glass\u2013ceramics in Italy was carried out. Original technogenic soils, classified as Spolic Technosols (ecotoxic),were pedotechnically in situ remediated by adding smectitic marlstone and Ca(OH)2. Chemical analysis on samples from piezometers showed the presence of harmful heavy metals in groundwater. By means of boreholes and soil profiles the newsoils generated, after remediation, were physically and chemically characterized and classified as Spolic Technosols (calcaric). Analysis on soil toxicity and leaching tests showed the effectiveness of the remediation and the mobility reduction of some potentially harmful elements according to the environmental Italian regulation