Fibronectin-binding protein B variation in Staphylococcus aureus

BACKGROUND: Fibronectin binding proteins A and B (FnBPA and FnBPB) mediate adhesion of S. aureus to fibrinogen, elastin and fibronectin. We previously identified seven different isotypes of FnBPA based on divergence in the fibrinogen- and elastin-binding A domains. The variation created differences in antigenicity while ligand binding functions were retained. Here, FnBPB variation was examined in both human and bovine isolates and compared to that of FnBPA. RESULTS: Seven different fnbB allelic variants were identified. Some strains that cluster by phylogenetic analysis contain different fnbB variants, whereas more divergent strains contain the same fnbB variant. The phylogeny of fnbB alleles does not match the phylogeny of fnbA alleles. Some FnBPA and FnBPB isotypes that are specified by human S. aureus strains are also found in bovine strains. The seven fnbB allelic variants encode seven distinct isotypes of the FnBPB A domain that are 61 to 85% identical in amino acid sequence. Variant amino acid residues were mapped on a three-dimensional model of the FnBPB A domain and were predicted to be surface-exposed. They are responsible for the antigenic diversity detected with polyclonal antibody and a monoclonal antibody raised against isotype I. Ligand binding by recombinant FnBPB N23 isotypes was compared by ELISA-based solid phase assays and surface plasmon resonance. Each bound to immobilized fibrinogen, elastin and fibronectin dose-dependently and saturably with similar affinities. Binding to fibronectin was surprising because the A domains do not contain any known motifs that mediate binding to fibronectin. This raises the possibility that the A domain of FnBPB contains a novel fibronectin binding motif that binds fibronectin by a novel mechanism. CONCLUSIONS: Seven different isoforms of FnBPB A domain retain ligand-binding functions but are antigenically distinct. The variation in FnBPA and FnBPB occurs in human and bovine S. aureus strains and may act as an immune evasion mechanism. All seven isotypes of FnBPB are capable of binding fibronectin though none contain any known fibronectin-binding motifs. These results have implications for the development of vaccines or immunotherapeutics that target FnBPB

Macroalgal forest restoration: the effect of the foundation species

IntroductionActive restoration is the strategy needed to trigger or accelerate the recovery of degraded marine habitats, which provide the goods and services essential for preserving biodiversity and human wellbeing. Ecosystem engineers are generally the target species in the ecological restoration of marine ecosystems, and large-sized brown macroalgae (e.g., the Cystoseira complex) are a priority due to their crucial ecological importance and vulnerability in many regions of the Mediterranean Sea. MethodsHere, we present the results of a successful intervention of ecological restoration of Gongolaria barbata. Results and DiscussionOne year after the restoration intervention, we observed the recovery of the canopy in terms of ca. 15m2 with a significant increase in faunal abundance and biodiversity compared with reference areas. However, despite the high restored macroalgal growth rates, the assemblage structure was still significantly different from that of reference pristine areas. Despite the fast faunal colonization of a new 3D habitat linked to the successful re-introduction of the ecosystem engineers (i.e., macroalgae), the complete restoration of the properties of a habitat can be a much longer process. We conclude that, after 1 year, active restoration of macroalgal forests still results in a partial recovery of ecosystem functions (i.e., rehabilitation) rather than a full ecosystem restoration, thus stressing the need for long-term monitoring of restoration interventions

Modeling macroalgal forest distribution at Mediterranean scale : present status, drivers of changes and insights for conservation and management

Macroalgal forests are one of the most productive and valuable marine ecosystems, but yet strongly exposed to fragmentation and loss. Detailed large-scale information on their distribution is largely lacking, hindering conservation initiatives. In this study, a systematic effort to combine spatial data on Cystoseira C. Agardh canopies (Fucales, Phaeophyta) was carried out to develop a Habitat Suitability Model (HSM) at Mediterranean scale, providing critical tools to improve site prioritization for their management, restoration and protection. A georeferenced database on the occurrence of 20 Cystoseira species was produced collecting all the available information from published and grey literature, web data portals and co-authors personal data. Data were associated to 55 predictor variable layers in the (ASCII) raster format and were used in order to develop the HSM by means of a Random Forest, a very effective Machine Learning technique. Knowledge about the distribution of Cystoseira canopies was available for about the 14% of the Mediterranean coastline. Absence data were available only for the 2% of the basin. Despite these gaps, our HSM showed high accuracy levels in reproducing Cystoseira distribution so that the first continuous maps of the habitat across the entire basin was produced. Misclassification errors mainly occurred in the eastern and southern part of the basin, where large gaps of knowledge emerged. The most relevant drivers were the geomorphological ones, followed by anthropogenic variables proxies of pollution and urbanization. Our model shows the importance of data sharing to combine a large number of spatial and environmental data, allowing to individuate areas with high probability of Cystoseira occurrence as suitable for its presence. This approach encourages the use of this modeling tool for the prediction of Cystoseira distribution and for supporting and planning conservation and management initiatives. The step forward is to refine the spatial information of presence-absence data about Cystoseira canopies and of environmental predictors in order to address species-specific assessments.peer-reviewe

Effects of Natural and Anthropogenic Stressors on Fucalean Brown Seaweeds Across Different Spatial Scales in the Mediterranean Sea

Este artículo contiene 14 páginas, 8 figuras, 3 tablas.Algal habitat-forming forests composed of fucalean brown seaweeds (Cystoseira, Ericaria, and Gongolaria) have severely declined along the Mediterranean coasts, endangering the maintenance of essential ecosystem services. Numerous factors determine the loss of these assemblages and operate at different spatial scales, which must be identified to plan conservation and restoration actions. To explore the critical stressors (natural and anthropogenic) that may cause habitat degradation, we investigated (a) the patterns of variability of fucalean forests in percentage cover (abundance) at three spatial scales (location, forest, transect) by visual estimates and or photographic sampling to identify relevant spatial scales of variation, (b) the correlation between semi-quantitative anthropogenic stressors, individually or cumulatively (MA-LUSI index), including natural stressors (confinement, sea urchin grazing), and percentage cover of functional groups (perennial, semi-perennial) at forest spatial scale. The results showed that impacts from mariculture and urbanization seem to be the main stressors affecting habitat-forming species. In particular, while mariculture, urbanization, and cumulative anthropogenic stress negatively correlated with the percentage cover of perennial fucalean species, the same stressors were positively correlated with the percentage cover of the semi-perennial Cystoseira compressa and C. compressa subsp. pustulata. Our results indicate that human impacts can determine spatial patterns in these fragmented and heterogeneous marine habitats, thus stressing the need of carefully considering scale-dependent ecological processes to support conservation and restoration.This study was supported by the European Union’s EASME (Executive Agency for Small and Medium Enterprise) and EMFF (European Maritime and Fisheries fund) as part of the project AFRIMED, “Algal Forest Restoration in the Mediterranean Sea” (under grant agreement no. 789059), http:// afrimed-project.eu/.Peer reviewe

The challenge of setting restoration targets for macroalgal forests under climate changes

Este artículo contiene 10 páginas, 5 figuras, 1 tabla.The process of site selection and spatial planning has received scarce attention in the scientific literature dealing with marine restoration, suggesting the need to better address how spatial planning tools could guide restoration interventions. In this study, for the first time, the consequences of adopting different restoration targets and criteria on spatial restoration prioritization have been assessed at a regional scale, including the consideration of climate changes. We applied the decision-support tool Marxan, widely used in systematic conservation planning on Mediterranean macroalgal forests. The loss of this habitat has been largely documented, with limited evidences of natural recovery. Spatial priorities were identified under six planning scenarios, considering three main restoration targets to reflect the objectives of the EU Biodiversity Strategy for 2030. Results show that the number of suitable sites for restoration is very limited at basin scale, and targets are only achieved when the recovery of 10% of regressing and extinct macroalgal forests is planned. Increasing targets translates into including unsuitable areas for restoration in Marxan solutions, amplifying the risk of ineffective interventions. Our analysis supports macroalgal forests restoration and provides guiding principles and criteria to strengthen the effectiveness of restoration actions across habitats. The constraints in finding suitable areas for restoration are discussed, and recommendations to guide planning to support future restoration interventions are also included.This study was funded by the EASME–EMFF (Sustainable Blue Economy) Project AFRIMED (http://afrimed-project.eu/, grant agreement N. 789059), supported by the European Community.Peer reviewe

Antibody-Based Agents in the Management of Antibiotic-Resistant Staphylococcus aureus Diseases

Staphylococcus aureus is a human pathogen that can cause a wide spectrum of diseases, including sepsis, pneumonia, arthritis, and endocarditis. Ineffective treatment of a number of staphylococcal infections with antibiotics is due to the development and spread of antibiotic-resistant strains following decades of antibiotic usage. This has generated renewed interest within the scientific community in alternative therapeutic agents, such as anti-S. aureus antibodies. Although the role of antibodies in the management of S. aureus diseases is controversial, the success of this pathogen in neutralizing humoral immunity clearly indicates that antibodies offer the host extensive protection. In this review, we report an update on efforts to develop antibody-based agents, particularly monoclonal antibodies, and their therapeutic potential in the passive immunization approach to the treatment and prevention of S. aureus infections

Staphylococcus aureus Manipulates Innate Immunity through Own and Host-Expressed Proteases

Neutrophils, complement system and skin collectively represent the main elements of the innate immune system, the first line of defense of the host against many common microorganisms. Bacterial pathogens have evolved strategies to counteract all these defense activities. Specifically, Staphylococcus aureus, a major human pathogen, secretes a variety of immune evasion molecules including proteases, which cleave components of the innate immune system or disrupt the integrity of extracellular matrix and intercellular connections of tissues. Additionally, S. aureus secretes proteins that can activate host zymogens which, in turn, target specific defense components. Secreted proteins can also inhibit the anti-bacterial function of neutrophils or complement system proteases, potentiating S. aureus chances of survival. Here, we review the current understanding of these proteases and modulators of host proteases in the functioning of innate immunity and describe the importance of these mechanisms in the pathology of staphylococcal diseases

Fluoride release and uptake abilities of conventonal glass ionomer cements affected by ultrasonic curing.

Objective: the objective of the study was to assess the effect of ultrasonic cur- ing on the fluoride release/uptake ca- pacities of two conventional glass ionomer cements (Fuji II and Fuji IX GP; GC Corporation, Tokyo, Japan). Methods : twenty discs of each materi- al were prepared and stored in deion- ized water at 37 °C. Two different cur- ing methods were investigated: stan- dard conditions and ultrasonic excita- tions. Cumulative fluoride release was meas- ured on days 1, 2, 3, 5, 7, 21, 35, and 49, then fluoride gel was applied to all the groups tested, and fluoride release (after reuptake) was measured on days 56, 70, 84, and 224 using a ion analyzer. Results : no significant differences were found between standard and ultrason- ic curing for Fuji II cement, whereas us- ing the Fuji IX GP cement a remarkable increase of fluoride release was report- ed when ultrasonic curing was per- formed. After exposure to fluoride gel, fluoride release increased significantly for all the groups tested. Significance: Ultrasonic curing does not affect fluoride release of Fuji II, whereas significantly increase fluoride release of Fuji IX GP

Purification of human plasma fibronectin using immobilized gelatin

This protocol describes a method for purification of fibronectin (Fn) from human plasma based on a combination of gel filtration and affinity chromatography steps. Clarified plasma is first loaded onto a Sepharose CL-4B column and unbound material is sequentially purified on columns containing covalently coupled gelatin and Arg. The elution conditions are optimized to obtain a homogeneous preparation of Fn on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Although the Fn yield is expected to be lower than that obtained using other methods, affinity adsorbents based on gelatin and Arg and gentle elution steps offer advantages including a high purity of the preparation and a correctly folded protein. The preparation can be useful for interaction studies and analysis of biological and immunological activities of Fn