ABSTRACT
BACKGROUND: Group B Streptococcus (GBS) is a commensal of healthy adults and an important pathogen in newborns, the elderly and immunocompromised individuals. GBS displays several virulence factors that promote colonisation and host infection, including the ST-17 strain-specific adhesin Srr2, previously characterised for its binding to fibrinogen. Another common target for bacterial adhesins and for host colonization is fibronectin, a multi-domain glycoprotein found ubiquitously in body fluids, in the extracellular matrix and on the surface of cells. RESULTS: In this study, fibronectin was identified as a novel ligand for the Srr2 adhesin of GBS. A derivative of the ST-17 strain BM110 overexpressing the srr2 gene showed an increased ability to bind fibrinogen and fibronectin, compared to the isogenic wild-type strain. Conversely, the deletion of srr2 impaired bacterial adhesion to both ligands. ELISA assays and surface plasmon resonance studies using the recombinant binding region (BR) form of Srr2 confirmed a direct interaction with fibronectin with an estimated Kd of 92 nM. Srr2-BR variants defective in fibrinogen binding also exhibited no interaction with fibronectin, suggesting that Srr2 binds this ligand through the dock-lock-latch mechanism, previously described for fibrinogen binding. The fibronectin site responsible for recombinant Srr2-BR binding was identified and localised in the central cell-binding domain of the protein. Finally, in the presence of fibronectin, the ability of a Δsrr2 mutant to adhere to human cervico-vaginal epithelial cells was significantly lower than that of the wild-type strain. CONCLUSION: By combining genetic and biochemical approaches, we demonstrate a new role for Srr2, namely interacting with fibronectin. We characterised the molecular mechanism of this interaction and demonstrated that it plays a role in promoting the adhesion of GBS to human cervico-vaginal epithelial cells, further substantiating the role of Srr2 as a factor responsible for the hypervirulence of GBS ST-17 strains. The discovery of the previously undescribed interaction between Srr2 and fibronectin establishes this adhesin as a key factor for GBS colonisation of host tissues.
Subject(s)
Adhesins, Bacterial , Bacterial Adhesion , Fibronectins , Protein Binding , Streptococcus agalactiae , Streptococcus agalactiae/genetics , Streptococcus agalactiae/metabolism , Streptococcus agalactiae/pathogenicity , Fibronectins/metabolism , Humans , Adhesins, Bacterial/metabolism , Adhesins, Bacterial/genetics , Fibrinogen/metabolism , Fibrinogen/genetics , Epithelial Cells/microbiology , Female , Streptococcal Infections/microbiology , Virulence Factors/metabolism , Virulence Factors/geneticsABSTRACT
Psychotropic drugs and benzodiazepines are nowadays among the primary substances of abuse. This results in a large and constant release into aquatic environments where they have potentially harmful effects on non-target organisms and, eventually, human health. In the last decades, evidence has been collected on the possible interference of benzodiazepines with reproductive processes, but data are few and incomplete. In this study, the possible negative influence of delorazepam on fertilization and embryo development has been tested in Paracentrotus lividus, a key model organism in studies of reproduction and embryonic development. Sperm, eggs, or fertilized eggs have been exposed to delorazepam at three concentrations: 1 µg/L (environmentally realistic), 5 µg/L, and 10 µg/L. Results indicate that delorazepam reduces the fertilizing capacity of male and female gametes and interferes with fertilization and embryo development. Exposure causes anatomical anomalies in plutei, accelerates/delays development, and alters the presence and distribution of glycoconjugates such as N-Acetyl-glucosamine, α-linked fucose, and α-linked mannose in both morulae and plutei. These results should attract attention to the reproductive fitness of aquatic species exposed to benzodiazepines and pave the way for further investigation of the effects they may exert on human fertility. The presence of benzodiazepines in the aquatic environment raises concerns about the reproductive well-being of aquatic species. Additionally, it prompts worries regarding potential impacts on human fertility due to the excessive use of anxiolytics.
Subject(s)
Paracentrotus , Male , Female , Animals , Humans , Benzodiazepines/adverse effects , Semen , Fertility , Fertilization , Embryo, NonmammalianABSTRACT
In recent decades, cadmium has emerged as an environmental stressor in aquatic ecosystems due to its persistence and toxicity. It can enter water bodies from various natural and anthropogenic sources and, once introduced into aquatic systems, can accumulate in sediments and biota, leading to bioaccumulation and biomagnification in the food chain. For this reason, the effects of cadmium on aquatic life remain an area of ongoing research and concern. In this paper, a multidisciplinary approach was used to assess the effects of long-term exposure to an environmental concentration on the hepatopancreas of farmed juveniles of sea bream, Sparus aurata. After determining metal uptake, metallothionein production was assessed to gain insight into the organism's defence response. The effects were also assessed by histological and ultrastructural analyses. The results indicate that cadmium accumulates in the hepatopancreas at significant concentrations, inducing structural and functional damage. Despite the parallel increase in metallothioneins, fibrosis, alterations in carbohydrate distribution and endocrine disruption were also observed. These effects would decrease animal fitness although it did not translate into high mortality or reduced growth. This could depend on the fact that the animals were farmed, protected from the pressure deriving from having to search for food or escape from predators. Not to be underestimated is the return to humans, as this species is edible. Understanding the behaviour of cadmium in aquatic systems, its effects at different trophic levels and the potential risks to human health from the consumption of contaminated seafood would therefore be essential for informed environmental management and policy decisions.
Subject(s)
Sea Bream , Water Pollutants, Chemical , Animals , Humans , Cadmium/toxicity , Sea Bream/physiology , Hepatopancreas , Ecosystem , Water Pollutants, Chemical/toxicityABSTRACT
Microplastics have become a great worldwide problem and it's therefore important to study their possible effects on human and environmental health. In this study, zebrafish embryos were used to compare two different sizes of polystyrene microplastics (PS-MPs), 1 µm and 3 µm respectively, at 0.01, 0.1, 1.0 and 10.0 mgL-1, and were monitored up to 72 h. Toxicity tests demonstrated that neither of the PS-MPs altered the embryos' survival and the normal hatching process. Instead, higher concentrations of both sizes caused an increase of the heart rate and phenotypic changes. The PS-MPs of both sizes entered and accumulated in the larvae at the concentration of 10.0 mgL-1 and the same concentration caused an increase of apoptotic processes correlated to redox homeostasis changes. The reported results give a realistic view of the negative effects of exposure to PS-MPs and provide new information on their toxicity, also considering their sizes.