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The intestinal lumen is a turbulent, semi-fluid landscape where microbial cells and nutrient-rich particles are distributed with high heterogeneity. Major questions regarding the basic physical structure of this dynamic microbial ecosystem remain unanswered. Most gut microbes are non-motile, and it is unclear how they achieve optimum localization relative to concentrated aggregations of dietary glycans that serve as their primary source of energy. In addition, a random spatial arrangement of cells in this environment is predicted to limit sustained interactions that drive co-evolution of microbial genomes. The ecological consequences of random versus organized microbial localization have the potential to control both the metabolic outputs of the microbiota and the propensity for enteric pathogens to participate in proximity-dependent microbial interactions. Here, we review evidence suggesting that several bacterial species adopt organized spatial arrangements in the gut via adhesion. We highlight examples where localization could contribute to antagonism or metabolic interdependency in nutrient degradation, and we discuss imaging- and sequencing-based technologies that have been used to assess the spatial positions of cells within complex microbial communities.

Bacterial biofilm formation and attachment to hosts are mediated by carbohydrate-binding lectins, exopolysaccharides, and their interactions in the extracellular matrix (ECM). During tomato infection Ralstonia pseudosolanacearum (Rps) GMI1000 highly expresses three lectins: LecM, LecF, and LecX. The latter two are uncharacterized. We evaluated the roles in bacterial wilt disease of LecF, a fucose-binding lectin, LecX, a xylose-binding lectin, and the Rps exopolysaccharide EPS I. Interestingly, single and double lectin mutants attached to tomato roots better and formed more biofilm under static conditions in vitro. Consistent with this finding, static bacterial aggregation was suppressed by heterologous expression of lecFGMI1000 and lecXGMI1000 in other Ralstonia strains that naturally lack these lectins. Crude ECM from a ΔlecF/X double mutant was more adhesive than the wild-type ECM, and LecF and LecX increased Rps attachment to ECM. The enhanced adhesiveness of the ΔlecF/X ECM could explain the double mutants hyper-attachment in static conditions. Unexpectedly, mutating lectins decreased Rps attachment and biofilm viscosity under shear stress, which this pathogen experiences in plant xylem. LecF, LecX, and EPS I were all essential for biofilm development in xylem fluid flowing through cellulose-coated microfluidic channels. These results suggest that under shear stress, LecF and LecX increase Rps attachment by interacting with the ECM and plant cell wall components like cellulose. In static conditions such as on root surfaces and in clogged xylem vessels, the same lectins suppress attachment to facilitate pathogen dispersal. Thus, Rps lectins have a dual biological function that depends on the physical environment.

Background: The interactions between bacterial pathogens and host cells are characterized by a multitude of complexities, leading to a wide range of heterogeneous outcomes. Despite extensive research, we still have a limited understanding of how bacterial motility in complex environments impacts their ability to tolerate antibiotics and adhere to mammalian cell surfaces. The challenge lies in unraveling the complexity of these interactions and developing quantitative microscopy approaches to predict the behavior of bacterial populations. Results: To address this challenge, we directed our efforts towards Pseudomonas aeruginosa, a pathogenic bacterium known for producing thick films in the lungs of cystic fibrosis patients, and Escherichia coli, used as a proof of concept to develop and demonstrate our single-cell tracking approaches. Our results revealed that P. aeruginosa exhibits diverse and complex interactions on mammalian cell surfaces, such as adhesion, rotational motion, and swimming, unlike the less interactive behavior of Escherichia coli. Our analysis indicated that P. aeruginosa demonstrated lower mean-squared displacement (MSD) values and greater adherence to mammalian cells compared to E. coli, which showed higher MSD slopes and less frequent adherence. Genetic mutations in membrane proteins of P. aeruginosa resulted in altered displacement patterns and reduced adhesion, with the ΔfliD mutant displaying a more Gaussian displacement distribution and significantly less adherence to mammalian cells. Adhesion and tolerance mechanisms are diverse and complex, potentially involving distinct pathways; however, our findings highlight the therapeutic potential of targeting the fliD gene (encoding a critical flagellum protein), as its deletion not only reduced adherence but also antibiotic tolerance. Conclusions: Overall, our findings underscore the importance of single cell tracking in accurately assessing bacterial behavior over short time periods and highlight its significant potential in guiding effective intervention strategies. [ABSTRACT FROM AUTHOR]

Catheter-associated urinary tract infections (CAUTIs) present significant health risks in medical settings, necessitating innovative solutions to prevent bacterial colonization on catheter surfaces. This study introduces a novel polymeric coating with dual antifouling and light-activated bactericidal properties to enhance the bactericidal efficacy of urinary catheters. The coatings were synthesized using a one-step process involving pyrogallol chemistry to deposit a copolymer composed of zwitterionic sulfobetaine for antifouling and sodium copper chlorophyllin, a photosensitizer that generates reactive oxygen species under light exposure to effectively kill bacteria. We evaluated the antifouling properties, cytocompatibility, and bactericidal performance of the coatings under various light conditions. The results showed significant reductions in bacterial adhesion, with light activation further endowing the catheter with bactericidal effects. Additionally, light could be delivered through an optical fiber within the catheter lumen to target and kill bacteria. The innovative coating using light-activated bactericidal action offers a promising approach to preventing CAUTIs, representing a potential breakthrough in developing safer and more effective urinary catheters. [ABSTRACT FROM AUTHOR]

The demand for dental implants has increased, establishing them as the standard of care for replacing missing teeth. Several factors contribute to the success or failure of an implant post-placement. Modifications to implant surfaces can enhance the biological interactions between bone cells and the implant, promoting better outcomes. Surface coatings have been developed to electrochemically alter implant surfaces, aiming to reduce healing time, enhance bone growth, and prevent bacterial adhesion. Quaternized silicon carbon nitride (QSiCN) is a novel material with unique electrochemical and biological properties. This study aimed to assess the influence of QSiCN, silicon carbide nitride (SiCN), and silicon carbide (SiC) coatings on the viability of osteoblast cells on nanostructured titanium surfaces. The experiment utilized thirty-two titanium sheets with anodized TiO2 nanotubes featuring nanotube diameters of 50 nm and 150 nm. These sheets were divided into eight groups (n = 4): QSiCN-coated 50 nm, QSiCN-coated 150 nm, SiCN-coated 50 nm, SiCN-coated 150 nm, SiC-coated 50 nm, SiC-coated 150 nm, non-coated 50 nm, and non-coated 150 nm. Preosteoblast MC3T3-E1 Subclone 4 cells (ATCC, USA) were used to evaluate osteoblast viability. After three days of cell growth, samples were assessed using scanning electron microscopy (SEM). The results indicated that QSiCN coatings significantly increased osteoblast proliferation (p < 0.005) compared to other groups. The enhanced cell adhesion observed with QSiCN coatings is likely due to the positive surface charge imparted by N+. [ABSTRACT FROM AUTHOR]

To evaluate the efficacy of the Aurie System, a preclinical prototype allowing for standardized intermittent catheter (IC) reuse of novel reusable no-touch ICs. Individuals with neurogenic bladder often require single-use ICs to urinate, but urinary tract infection (UTI) is a common cause of morbidity for IC users. Safer no-touch catheters are not easily affordable, and the Aurie System attempts to provide no-touch catheters at a fraction of the price by allowing for standardized and safe IC reuse. Standard ICs were inoculated with Escherichia coli and Pseudomonas aeruginosa and incubated for 48 hours to assess microbial burden and biofilm formation (the latter using infrared fluorescence imaging). This procedure was repeated with Aurie ICs, focusing on evaluating catheter microbial burden after inoculation and reprocessing with the prototype washer-disinfector. This was repeated with up to 100 cycles to evaluate repetitive use. Standard ICs showed bacterial attachment and biofilm development peaking at 24 hours of incubation. The Aurie catheters produced a similar outcome but, after reprocessing, microbial burden was reduced below the level of detection. Repeat cycles showed pathogen clearance to similar levels. One catheter reached 100 cycles and there was no viable pathogen load after reprocessing. Intermittent urinary catheters, when cleaned inappropriately, can harbor viable bacteria and biofilm. The Aurie System, when used to disinfect novel reusable ICs within a prototype reprocessing device, can reduce microbial burden below level of detection even after 100 cycles. This suggests the Aurie System may be a feasible technology for safe IC reuse. [ABSTRACT FROM AUTHOR]

Medical device-associated infections are a pervasive global healthcare concern, often leading to severe complications. Bacterial biofilms that form on indwelling medical devices, such as catheters, are significant contributors to infections like bloodstream and urinary tract infections. This study addresses the challenge of biofilms on medical devices by introducing a portable antimicrobial catheter insert (PACI) designed to be efficient, biocompatible, and anti-infective. The PACI utilizes nitric oxide (NO), known for its potent antimicrobial properties, to deter bacterial adhesion and biofilm formation. To achieve this, a photoinitiated NO donor, S -nitroso- N -acetylpenicillamine (SNAP), is covalently linked to a polydimethylsiloxane (PDMS) polymer. This design allows for higher NO loading for long-term impact and prevents premature donor leaching, a common challenge with SNAP-blended polymers. The SNAP-PDMS material was applied to a side-glowing fiber optic and connected to a wearable light module emitting 450 nm light, creating a functional antimicrobial insert. Activation of the fiber optic, accomplished with a one-click mechanism, enables real-time NO release, maintaining controlled NO levels for a minimum of 24 hours. The therapeutic levels of NO released via photocatalysis from the PACI demonstrated remarkable efficacy, with >90 % reduction in bacterial viability against S. aureus , S. epidermidis , and P. mirabilis without any cytotoxic impact on mammalian cells. This study underscores the potential of the NO-releasing insert in clinical settings, providing a portable and adaptable solution for preventing catheter-associated infections. Photoactivated nitric oxide (NO)-release levels can be achieved using a wireless and Portable Antimicrobial Catheter Insert (PACI) at the medical device interface. The covalent attachment of the NO donor to the base PDMS polymer enables higher NO loading and prevents premature leaching, ensuring maximum efficiency in eradicating catheter-associated infections. [Display omitted] [ABSTRACT FROM AUTHOR]

This study is on fly ash (FA) concrete admixed with TiO2 nanoparticles (FAT), CaCO3 nanoparticles (FAC), and an equal ratio of TiO2 and CaCO3 nanoparticles (FATC) were exposed to seawater for 365 days in order to find the attachment of macrofoulants on their surface. The pH reduction studies were carried out in order to determine concrete deterioration in seawater, the results showed that FA specimens had more pH reduction than FAT, FAC and FATC. Total viable count, epifluorescence microscopy and denaturing gradient gel electrophoresis (DGGE) analysis confirmed the intensity of bacterial attachment and its diversity on FA, FAT, FAC, and FATC specimens. A systematic and comparative analysis predicted the overall relationship in terms of the dominant bacterial species on the different fly ash concrete specimens. To determine the biodeterioration of the fly ash concrete specimens in seawater, we used thermography experiments to evaluate the regions affected by bacteria on mortar specimens and confirmed that FA and FAC had more degradation and that FAT and FATC were the least degraded. Laser Raman Spectroscopy was interesting to find the white micron-sized particles of sulphur on FAC, FAT, and FATC specimens after 365 days immersed in seawater. We used a confocal laser scanning microscope to estimate the thickness of biofilm growth on FAT (13.91 0.38 mm), FATC (21.64 0.22 mm), FAC (33.56 0.26 mm), and FA (43.36 0.10 mm) specimens. The results showed that FAT and FATC specimens were the superior specimens, with enhanced biofouling and biodeterioration resistance in a seawater environment. Practical Applications: Fly ash is a waste material from thermal power plants and is used as a supplementary cementitious material in the construction industry. However, fly ash concrete has a slow hydration process and early strength problems. To overcome these problems, we used TiO2 and CaCO3 nanoparticles to enhance the properties of fly ash concrete. Nanoparticles have a high surface area and unique functional properties such as maintaining concrete pH, reducing porosity, enhancing pozzolanic activity, and so forth. In this study, we used TiO2 and CaCO3 nanoparticles to improve the strength of fly ash concrete and provide it with antibacterial properties. This study is significant in advancing our knowledge of fly ash concrete modified with nanoparticles and exposed to seawater with respect to its mechanical properties, durability, and antibacterial properties. Addition of nanoparticles upgraded the fly ash concrete towards its practical applications. [ABSTRACT FROM AUTHOR]

Nosocomial infections affect implanted medical devices and greatly challenge their functional outcomes, becoming sometimes life threatening for the patients. Therefore, aggressive antibiotic therapies are administered, which often require the use of last‐resort drugs, if the infection is caused by multi‐drug‐resistant bacteria. Reducing the risk of bacterial contamination of medical devices in the hospitals has thus become an emerging issue. Promising routes to control these infections are based on materials provided with intrinsic bactericidal properties (i.e., chemical action) and on the design of surface coatings able to limit bacteria adhesion and fouling phenomena (i.e., physical action), thus preventing bacterial biofilm formation. Here, we report the development and validation of coatings made of layer‐by‐layer deposition of electrospun poly(vinylidene fluoride‐co‐trifluoro ethylene) P(VDF‐TrFE) fibers with controlled orientations, which ultimately gave rise to antifouling surfaces. The obtained 10‐layer surface morphology with 90° orientation fibers was able to efficiently prevent the adhesion of bacteria, by establishing a superhydrophobic‐like behavior compatible with the Cassie‐Baxter regimen. Moreover, the results highlighted that surface wettability and bacteria adhesion could be controlled using fibers with diameter comparable to bacteria size (i.e., achievable via electrospinning process), by tuning the intra‐fiber spacing, with relevant implications in the future design of biomedical surface coatings. [ABSTRACT FROM AUTHOR]

Intravenous (IV) catheter-related bloodstream infections (CRBSIs) cause significant risks in healthcare, necessitating advancements in catheter design and materials. This study investigates the effectiveness of Ecoflex, a silicone-based material, in studying CRBSIs through the development of skin-like replicas that mimic human skin properties for use in wearable sensing devices. We characterized the replica's bioinspired surface roughness, wettability, bacterial adhesion, and mechanical properties and validated its performance using in vitro IV simulation. The results demonstrated that the bioinspired model replicates human skin textures with less than 7.5% error for surface roughness ranging from 0.05 μm to 6.3 μm. Wettability tests revealed that the artificial sebum application significantly reduced the static contact angles for deionized water and artificial sweat. Comprehensive mechanical testing revealed material high elasticity and resilience, suitable for dynamic biomedical applications. Bacterial adhesion studies using Staphylococcus epidermidis showed varying adhesion patterns influenced by surface roughness, highlighting the potential for material texture to impact infection risk. In IV therapy simulations, we observed bacterial growth dynamics over the incubation period. Our findings suggest that Ecoflex-based skin-like replicas can serve as a valuable tool for developing and testing new catheters, while the potential for use in other medical innovation devices, including wearable sensing devices, ultimately contributes to improved patient outcomes and infection control strategies. [ABSTRACT FROM AUTHOR]

Type IV pili (T4P) produced by the pathogen Pseudomonas aeruginosa play a pivotal role in adhesion, surface motility, biofilm formation, and infection in humans. Despite the significance of T4P as a potential therapeutic target, key details of their dynamic assembly and underlying molecular mechanisms of pilus extension and retraction remain elusive, primarily due to challenges in isolating intact T4P machines from the bacterial cell envelope. Here, we combine cryo-electron tomography with subtomogram averaging and integrative modelling to resolve in-situ architectural details of the dynamic T4P machine in P. aeruginosa cells. The T4P machine forms 7-fold symmetric cage-like structures anchored in the cell envelope, providing a molecular framework for the rapid exchange of major pilin subunits during pilus extension and retraction. Our data suggest that the T4P adhesin PilY1 forms a champagne-cork-shaped structure, effectively blocking the secretin channel in the outer membrane whereas the minor-pilin complex in the periplasm appears to contact PilY1 via the central pore of the secretin gate. These findings point to a hypothetical model where the interplay between the secretin protein PilQ and the PilY1-minor-pilin priming complex is important for optimizing conformations of the T4P machine in P. aeruginosa, suggesting a gate-keeping mechanism that regulates pilus dynamics. Type IV pili (T4P) enable bacteria to sense, move, and adhere to surfaces. Here, the authors solve the structure of the T4P machine in Pseudomonas aeruginosa, revealing the localization and regulatory role of its tip adhesin, PilY1. [ABSTRACT FROM AUTHOR]

Aim Methodology Results Conclusions To evaluate the effect of chlorhexidine gluconate‐loaded phase‐transited lysozyme (CHG@PTL) coating on inhibiting bacterial adhesion and biofilm formation in an ex vivo root canal dentine model.The physicochemical and structural characteristics of CHG@PTL nanoparticle suspension and its coating formed on the dentine surface were analysed by thioflavin T fluorescence assay, transmission electron microscopy and confocal laser scanning microscopy (CLSM). The sustained chlorhexidine release profile of the CHG@PTL coating on the dentine surface was compared with that of the 2% CHG solution. By comparing with phosphate‐buffered saline, 1% sodium hypochlorite and 2% CHG solutions, the sustained antibacterial ability of the CHG@PTL coating and its effects on adhesion and biofilm formation of three types of bacteria (E. faecalis, S. mutans, and A. viscous) were analysed in ex vivo root canal dentine models using the serial plate transfer test (SPTT) and CLSM with live/dead bacterial staining, respectively.CHG promoted the lysozyme protein to form a higher proportion of β‐sheet structure during phase transition. In the CHG@PTL nanoparticle suspension, characteristic drug‐loaded nanospheres with a high concentration of CHG molecules inside and an outer PTL nanofilm were observed, and they formed a thinner and tighter coating on the dentine surface. The CHG@PTL coating on the dentine surface showed a significantly higher cumulative release amount of chlorhexidine than that of 2% CHG (p < .05). The results of SPTT showed that the CHG@PTL coating had a longer antibacterial duration than the control groups. After 12 h of incubation, a higher number of bacteria were agglutinated on the CHG@PTL coating surface compared to the control groups (p < .05). After 7 days of incubation, the number of agglutinated bacteria significantly decreased. At two time points, the percentage of dead bacteria on the CHG@PTL coating surface was the highest among all experimental groups based on CLSM observation (over 99.9% for all three bacteria, p < .001).CHG@PTL nanoparticle suspension could form an antimicrobial coating on the surface of dentine with a novel ‘agglutinating bacteria and sterilizing’ mode. [ABSTRACT FROM AUTHOR]

Glyceraldehyde‐3‐phosphate‐dehydrogenase (GAPDH; EC1.2.1.12) has several functions in Mycobacterium tuberculosis (Mtb) and the human host. Apart from its role in glycolysis, it serves both as a cell surface and a secreted receptor for plasmin(ogen) (Plg/Plm), transferrin (Tf), and lactoferrin (Lf). Plg sequestration by Mtb GAPDH facilitates bacterial adhesion and tissue invasion, while an equivalent interaction with host GAPDH regulates immune cell migration. In both, host and microbe, internalization of Tf/Lf‐GAPDH complexes serves as a route for iron acquisition. To date, the structure of Mtb GAPDH or the residues involved in these moonlighting interactions have not been identified. This study provides the first known X‐ray crystal structure of Mtb GAPDH. Through further mutagenesis and functional assays, we found that the C‐terminal lysines of Mtb and human GAPDH affect enzyme activity and ligand binding. We also establish the stoichiometry of Plg, Tf and Lf interactions with the GAPDH tetramer. Lastly, molecular simulation studies reveal the interactions of the C‐terminal lysine residues. [ABSTRACT FROM AUTHOR]

The growing scarcity of clean water necessitates urgent interventions in wastewater treatment. Polymeric membranes are prevalent in this domain. However, due to their hydrophobic behaviour, they become more prone to fouling. To address this issue, we synthesized hydrophilic nanomaterials as additives to prepare polymer nanocomposite membranes with enhanced hydrophilicity. The eco-friendly zinc oxide (ZnO) nanomaterial was synthesized via a green route, and reduced graphene oxide (rGO) and a rGO/ZnO composite were synthesized using a chemical process and characterized using FESEM, FTIR, XRD, and zeta potential techniques. The synthesized nanomaterials were incorporated into PVDF membranes, and the prepared membranes were characterized using FESEM with EDX, FTIR, and contact angle measurements, along with mechanical strength determination. The filtration properties of the membranes were evaluated using a dead-end filtration module. The FESEM images revealed the finger-like structure in the membranes, which increases with the incorporation of nanomaterials. The results demonstrated that among all the membranes, the rGO/ZnO-PVDF showed the highest membrane permeability due to increased membrane porosity, pore size, and finger-like macrovoid sponge structures. The rejection of BSA protein and CBB dye increased due to the negative charge provided by the ZnO in the rGO/ZnO nanocomposite. The antifouling study reveals that the rGO/ZnO-PVDF membrane had the highest FRR value of ∼50.05% among the various synthesized membranes. The disk diffusion data of the synthesized nanomaterials and nanocomposite membranes confirms their antibacterial activity and suggests that the addition of rGO/ZnO nanomaterial can provide anti-biofouling behaviour to the polymer membranes by minimizing the adhesion of bacteria on the surface of the membrane. The results showed that the rGO/ZnO-PVDF membrane possesses antifouling and anti-biofouling behaviour, useful for minimising the pollutants from wastewater. [ABSTRACT FROM AUTHOR]

Background: Dental restorative materials are recognized as artificial niches that facilitate the adherence and accumulation of oral microorganisms. To mitigate oral diseases and extend the lifespan of restorations, it is advantageous to use dental materials that exhibit low susceptibility to bacterial adhesion. Objective: To evaluate and compare bacterial adhesion on two bioactive restorative materials, a glass hybrid restorative, and an alkasite with a nanohybrid resin composite as a positive control. The secondary objectives were to compare the surface roughness (SR) of the materials and determine the correlation between the bacterial adhesion and the SR. Materials and methods: The samples consisted of 33 polished discs of each material: Group A: Tetric® N-Ceram (nanohybrid resin composite), Group B: Equia Forte™ HT Fil (glass hybrid restorative) and Group C: Cention N® (alkasite). Streptococcus mutans cultures were inoculated and after 24-hours of incubation, bacterial adhesion was measured by measuring optical density (OD) and number of colony forming units (CFUs). After 96-hours incubation, the bacterial cell count was determined using scanning electron microscopy (SEM). SR was assessed using surface profilometer. Results: Alkasite had significantly lower OD and CFUs (p < 0.001 and p = 0.015 respectively). According to the SEM analysis, the glass hybrid restorative had lower mean bacterial cell count with no significant difference between the groups. The nanohybrid composite had the smoothest surface that was significantly lower than the alkasite and glass hybrid restorative (p = 0.002). None of the groups demonstrated a correlation between bacterial adhesion and SR. Conclusion: Alkasite impedes bacterial adhesion better than the glass hybrid restorative and nanohybrid composite, while smoother surfaces are achieved with the nanohybrid composite. [ABSTRACT FROM AUTHOR]

Porous zirconia scaffolds manufactured using polymer-infiltrated ceramic network (PICN) and additive manufacturing technologies are emerging as promising alternatives to traditional ceramic materials in dental restorations. However, incomplete osseointegration and bacterial infections still represent challenges for the long-term performance of this new composite material. To address this, the present study aims to investigate the effect of peptide biofunctionalization on the biological performance of infiltrated zirconia scaffold surfaces. The samples used in the work consisted of a 3D-printed zirconia scaffold infiltrated with a dimethacrylate copolymer. Surface biofunctionalization was achieved using a synthetic platform containing the cell-adhesive sequence RGD and the antibacteria LF1-11 peptide (RGD-LF). The attachment of the molecule was characterized through fluorescence confocal laser scanning microscopy and X-ray photoelectron spectroscopy. The biological performance of the samples was evaluated in terms of human mesenchymal stem cell adhesion and early attachment of S. aureus. The physicochemical characterization verified the successful anchoring of the biomolecule to the surface, leading to a peptide density of 288 pmol/cm2. The biological assays confirmed the potential of RGD-LF to improve cell adhesion and spreading. In this sense, the average cell area increased fourfold in the biofunctionalized surface. Regarding bacterial adhesion, it was demonstrated that RGD-LF significantly inhibited it, reducing early adhesion by half compared to the untreated surface. Overall, this study provides valuable insights into the biofunctionalization of polymer-infiltrated 3D scaffolds for the development of cell-instructive and antibacterial surfaces tailored for dental applications. [ABSTRACT FROM AUTHOR]

Dentin hypersensitivity (DH) manifests as sharp and uncomfortable pain due to the exposure of dentinal tubules (DTs) following the erosion of tooth enamel. Desensitizing agents commonly used in clinical practice have limitations such as limited depth of penetration, slow remineralization and no antimicrobial properties. To alleviate these challenges, our study designed a lactoferrin-derived amyloid nanofilm (PTLF nanofilm) inspired by the saliva-acquired membrane (SAP). The nanofilm utilises Tris(2-carboxyethyl)phosphine (TCEP) to disrupt the disulfide bonds of lactoferrin (LF) under physiological conditions. The PTLF nanofilm modifies surfaces across various substrates and effectively prevents the early and stable adhesion of cariogenic bacteria, such as Streptococcus mutans and Lactobacillus acidophilus. Simultaneously, it adheres rapidly and securely to demineralized dentin surfaces, facilitating in-situ remineralization of HAP through a simple immersion process. This leads to the formation of a remineralized layer resembling natural dentin, with an occlusion depth of dentinal tubules exceeding 80 µm after three days. The in vivo and vitro results confirm that the PTLF nanofilm possesses good biocompatibility and its ability to exert simultaneous antimicrobial effects and dentin remineralization. Accordingly, this innovative bifunctional PTLF amyloid coating offers promising prospects for the management of DH-related conditions. 1. We design a simple, fast, inexpensive, and easy-to-process PTLF nanofilm for nearly any material surface or shape. 2. The PTLF nanofilm modifies surfaces across various substrates and effectively prevents the adhesion of cariogenic bacteria, such as Streptococcus mutans and Lactobacillus acidophilus. 3. The abundant functional groups on the surface of PTLF nanofilm facilitate bioactive hydroxyapatite (HAP) formation and maintain stability at the HAP remineralization interface. [Display omitted] [ABSTRACT FROM AUTHOR]

The Riemerella anatipestifer bacterium is known to cause infectious serositis in ducklings. Moreover, its adherence to the host's respiratory mucosa is a critical step in pathogenesis. Membrane cofactor protein (MCP; CD46) is a complement regulatory factor on the surface of eukaryotic cell membranes. Bacteria have been found to bind to this protein on host cells. Outer membrane proteins (OMPs) are necessary for adhesion, colonisation, and pathogenicity of Gram-negative bacteria; however, the mechanism by which R. anatipestifer adheres to duck cells remains unclear. In this study, pull-down assays and LC–MS/MS identified eleven OMPs interacting with duck CD46 (dCD46), with OMP71 exhibiting the strongest binding. The ability of an omp71 gene deletion strain to bind dCD46 is weaker than that of the wild-type strain, suggesting that this interaction is important. Further evidence of this interaction was obtained by synthesising OMP71 using an Escherichia coli recombinant protein expression system. Adhesion and invasion assays and protein and antibody blocking assays confirmed that OMP71 promoted the R. anatipestifer YM strain (RA-YM) adhesion to duck embryo fibroblasts (DEFs) by binding to CD46. Tests of the pathogenicity of a Δomp71 mutant strain of RA-YM on ducks compared to the wild-type parent supported the hypothesis that OMP71 was a key virulence factor of RA-YM. In summary, the finding that R. anatipestifer exploits CD46 to bind to host cells via OMP71 increases our understanding of the molecular mechanism of R. anatipestifer invasion. The finding suggests potential targets for preventing and treating diseases related to R. anatipestifer infection. [ABSTRACT FROM AUTHOR]

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Urinary tract infections (UTIs) affect nearly 50% of women in their lifetime. Uropathogenic Escherichia coli (UPEC) expresses F9/Fml pili tipped with the protein FmlH that specifically bind to terminal galactoside and galactosaminoside units in glycoproteins on kidney and bladder cells and colonize host tissues. The traditional UTI treatment using excessive antibiotics has led to the rise in various UPEC antibiotic-resistant strains. An alternative therapeutic approach prevents the initial bacterial attachment on the host cells using competitive FmlH-binding inhibitors. In this study, we used computer-aided drug design techniques to identify novel glycomimetics that are predicted to bind strongly to and inhibit the UPEC FmlH. We performed in silico receptor-based and ligand-based scaffold hopping, and molecular docking to predict novel FmlH-binding glycomimetics with high chemical synthesizability. We replaced the two major scaffolds of the most potent known FmlH-binding ligand to obtain novel compounds. Additionally, we applied global machine-learning models to predict the ADMET properties of the molecules. Compounds with low ADMET risks were subjected to molecular dynamics simulations and a detailed investigation of the FmlH–glycomimetic interactions was performed. We have prepared and supplied a library of 58 novel glycomimetics that can be subjected to further biological activity studies. [ABSTRACT FROM AUTHOR]

This study explores and verifies potential molecular targets through which KRAS mutations regulate the colonization of Fusobacterium nucleatum (FN) in colorectal cancer (CRC). This study combined multiple bioinformatics methods and biological assays. Through The Cancer Genome Atlas, Gene Expression Omnibus, Human Protein Atlas, immunohistochemistry, and co‐culture assays, we further confirmed the differential expression of SERTAD4 in CRC. We delved deeper into examining how expression of SERTAD4 is linked with immune cell infiltration and the enrichment of potential pathways. Lastly, through bacterial phenotypic assays, we validated the function of SERTAD4. As a molecule associated with KRAS mutations and FN infection, the expression levels of SERTAD4 were downregulated in CRC. The diagnostic efficacy of SERTAD4 for CRC is not inferior to that of CEA. Low expression of SERTAD4 is associated with poorer overall survival in CRC. Correlation analysis found that increased expression of SERTAD4 is associated with various immune cell infiltrations and immune checkpoint genes. Finally, bacterial adhesion and invasion assays verify that SERTAD4 inhibits the adhesion and invasion abilities of FN in CRC. This study demonstrates that SERTAD4 exerts a protective role in CRC by inhibiting the colonization of FN. [ABSTRACT FROM AUTHOR]

Ventilator-associated pneumonia (VAP) remains one of the most common hospital-acquired infections (HAI). Considering the complicated diagnosis and the lack of effective treatment, prophylactic measures are suggested as the new standard to prevent the disease. Although VAP often manifests a polymicrobial nature, Pseudomonas aeruginosa remains one of the pathogens associated with the highest morbidity and mortality rates within these mechanically ventilated patients. In this paper, we report on the development of an antibacterial hydrogel coating using the polymyxin B (PMB) peptide to prevent bacterial adhesion to the polymeric substrate. We fully characterized the properties of the coating using atomic force microscopy (AFM), scanning electron microscopy (SEM), wettability analyses and Fourier-transform infrared (FTIR) and Raman spectroscopy. Furthermore, several biological assays confirmed the antibacterial and anti-biofilm effect of the tubing for at least 8 days against P. aeruginosa. On top of that, the produced coating is compliant with the requirements regarding cytocompatibility stated in the ISO (International Organization for Standardization) 10993 guidelines and an extended release of PMB over a period of at least 42 days was detected. In conclusion, this study serves as a foundation for peptide-releasing hydrogel formulas in the prevention of VAP. [ABSTRACT FROM AUTHOR]

The colonization of microbes and the resulting formation of biofilms on dental implants are significant contributors to peri-implantitis and the failure of these implants. The aim of the research was to analyze the impact of density and depth of laser texturing of the Ti-6Al-7Nb alloy surface on the colonization of selected microorganisms and biofilm formation. Standard strains of Gram-negative and Gram-positive bacteria and yeasts from the American Type Culture Collection—ATCC—were used to demonstrate the ability to form single-species biofilms in vitro. The study evaluated three types of titanium samples with different texture density and depth. The colonization and biofilm formation abilities of the tested microorganisms were assessed. The obtained results were subjected to statistical analysis. Among the analyzed strains, L. rhamnosus showed the highest colonization of the tested surfaces. It was found that there is no relationship between the texture parameters and the number of colony-forming units (CFU/mL) for C. albicans, S. mutans, and L. rhamnosus. For the F. nucleatum strain, it was shown that the number of colony-forming bacteria is related to the texture density. [ABSTRACT FROM AUTHOR]

The Gramnegative bacterium Myxococcus xanthus glides on solid surfaces. Dynamic bacterial focal adhesion complexes (bFACs) convert proton motive force from the inner membrane into mechanical propulsion on the cell surface. It is unclear how the mechanical force transmits across the rigid peptidoglycan (PG) cell wall. Here, we show that AgmT, a highly abundant lytic PG transglycosylase homologous to Escherichia coli MltG, couples bFACs to PG. Coprecipitation assay and singleparticle microscopy reveal that the gliding motors fail to connect to PG and thus are unable to assemble into bFACs in the absence of an active AgmT. Heterologous expression of E. coli MltG restores the connection between PG and bFACs and thus rescues gliding motility in the M. xanthus cells that lack AgmT. Our results indicate that bFACs anchor to AgmTmodified PG to transmit mechanical force across the PG cell wall. [ABSTRACT FROM AUTHOR]

As the habitats of bacteria, soil pore network and surface properties control the distribution, adhesion, and motility of bacteria in soils. These physical processes in turn influence bacterial accesses to nutrients and bacterial interactions. Our understanding on the pore- and surface-mediated bacterial interactions is currently limited. In this research, we evaluated the effects of soil pore confinement and surface adhesion on conjugation-based bacterial interactions. The interaction was measured by plasmid transfer between donor and recipient cells within the population of soil bacterium Pseudomonas putida. We found that the presence of porous sand media led to a net increase in conjugation frequency compared to sand-free liquid control. The increase is attributed to the facilitated effect of pore confinement on the collision of bacteria within pores. In contrast, bacterial adhesion to sand surfaces under elevated ionic strength conditions decreased the conjugation frequency as a result of mobility reduction on the surface. Such collision and adhesion mechanisms jointly drive the conjugation as a function of pore and surface properties of porous media. These results provide valuable insights into the roles of soil pores and surfaces in regulating horizontal gene transfer, an essential cell-to-cell interaction sustaining key processes of soil ecology and health. [ABSTRACT FROM AUTHOR]

Polymers are versatile materials widely used in various industries, with significant applications in biomedicine where biofouling on polymer surfaces presents major health and economic challenges. Biofouling, initiated by bacterial adhesion, can be mitigated by modifying surface properties through laser micro- and nano-texturing, an approach that offers advantages over chemical treatments. This study introduces an economical mass production process for textured polymeric components using injection molding to replicate hierarchical textures. Testing revealed that all textured samples significantly reduced bacterial adhesion compared to untextured surfaces across different designs and bacteria types after 24 h of culture. The study examined factors like wettability, nanoscale roughness, and pattern dimensions to explain these outcomes, comparing them with existing studies. Despite all textured samples showing decreased wettability and roughness, these factors alone did not ensure reduced bacterial adhesion. The most effective anti-adhesive performance was observed in surfaces with parallel ridge patterns, which segmented the surface into isolated areas that limited bacterial interaction and hindered micro-colony formation, highlighting the importance of specific surface patterning in combating biofouling. [ABSTRACT FROM AUTHOR]

In cattle, bovine respiratory syncytial virus (BRSV) is associated with secondary bacterial infections; however, the mechanisms of the interaction between BRSV and bacteria are unclear. Trueperella pyogenes (T. pyogenes) causes pneumonia in cattle and is involved in secondary infections following viral infections. In this study, we evaluated the effect of BRSV infection on the adhesion of T. pyogenes to BRSV-infected cells. BRSV infection significantly enhanced the adhesion of T. pyogenes to cells in a multiplicity of infection- and time-dependent manner. The BRSV-mediated change in the adhesion of T. pyogenes was widely observed in various cell types and bacterial strains. The results from the gentamicin protection assay showed that BRSV infection did not affect the intracellular invasion ability of T. pyogenes. Furthermore, adhesion assays conducted using BRSV G protein-expressing cells and anti-BRSV G antibodies revealed that the increased adhesion of T. pyogenes to cells was mediated by the G protein of BRSV. In addition, immunofluorescence assay revealed the colocalization of BRSV G protein and T. pyogenes. Thus, BRSV infection can potentially lead to bovine respiratory disease complex by promoting the adhesion of T. pyogenes to the infected cells. [ABSTRACT FROM AUTHOR]

The research exploiting the ability of dental materials to induce or prevent secondary caries (SC) development still seems inconclusive. Controlling bacterial adhesion by releasing bacteriostatic ions and improving the surface structure has been suggested to reduce the occurrence of SC. This paper analyses the impact of five distinctively composed dental materials on cariogenic biofilm formation. Forty-five specimens of three composites (CeramX Spectra ST, Admira Fusion, Beautifil II) and two glass–ionomers (Fuji II LC, Caredyne Restore), respectively, were incubated in bacterial suspension composed of Streptococcus mutans, Lactobacillus acidophilus, Streptococcus mitis, Streptococcus sanguinis, and Streptococcus salivarius at pH 7.0 and 5.5. Coverslips were used as a control. Adhered bacteria were collected after 2, 4, 6, 12, 24, and 48 h and analyzed using quantitative polymerase chain reaction (qPCR). Fluoride leakage was measured at each collection. The specimens' surface topography was assessed using interferometry. In the present study, surface roughness seemed to have a partial role in bacterial adhesion and biofilm formation, together with chemical composition of the materials tested. Despite differences in fluoride leakage, biofilm accumulation was similar across materials, but the number of adhered bacteria differed significantly. A release of other ions may also affect adhesion. These variations suggest that certain materials may be more prone to initiating secondary caries. [ABSTRACT FROM AUTHOR]

During implant maintenance, preserving a smooth surface on the machined transmucosal abutment is critical to reduce biofilm attachment and colonization. The present study compared the surface roughness and bacterial colonization of machined titanium surfaces after instrumentation with various materials. Forty-four machined grade 23 titanium discs were instrumented with a round polyether ether ketone (PEEK) tip, a plastic curette tip, or a pure titanium curette tip with piezoelectric devices. Before and after instrumentation, the surface roughness (Ra and Rz) values were analyzed with a profilometer and scanning electron microscopy (SEM). Streptococcus sanguinis was cultured and incubated for 24 hours on the instrumented discs, and colony-forming units per milliliter were obtained for each group. Samples instrumented with the metal ultrasonic tip significantly increased surface roughness compared with the other groups. This resulted in greater colonization by S. sanguinis than surfaces instrumented with PEEK tips or the negative control. Samples instrumented with PEEK and plastic tips did not exhibit any statistically significant increase in surface roughness, and SEM analysis revealed a significantly rougher surface of discs instrumented with metal compared with discs instrumented with plastic or PEEK tips despite the possibility of debris from tip dissolution. Our results suggest that instrumentation with metal ultrasonic tips with piezoelectric devices significantly increased machined titanium's surface roughness and elicited higher biofilm formation in vitro. Meanwhile, instrumentation of machined titanium with PEEK or plastic ultrasonic tips did not affect the surface roughness or bacterial adhesion. [ABSTRACT FROM AUTHOR]

Objectives: This study investigates the impact of nutrient availability on the growth, adhesion, and biofilm formation of Pseudomonas aeruginosa ATCC 27853 under static conditions. Methods: Bacterial behaviour was evaluated in nutrient-rich Luria–Bertani (LB) broth and nutrient-limited M9 media, specifically lacking carbon (M9-C), nitrogen (M9-N), or phosphorus (M9-P). Bacterial adhesion was analysed microscopically during the transition from reversible to irreversible attachment (up to 120 min) and during biofilm production/maturation stages (up to 72 h). Results: Results demonstrated that LB and M9 media supported bacterial growth, whereas nutrient-starved conditions halted growth, with M9-C and M9-N inducing stationary phases and M9-P leading to cell death. Fractal analysis was employed to characterise the spatial distribution and complexity of bacterial adhesion patterns, revealing that nutrient-limited conditions affected both adhesion density and biofilm architecture, particularly in M9-C. In addition, live/dead staining confirmed a higher proportion of dead cells in M9-P over time (at 48 and 72 h). Conclusions: This study highlights how nutrient starvation influences biofilm formation and bacterial dispersion, offering insights into the survival strategies of P. aeruginosa in resource-limited environments. These findings should contribute to a better understanding of biofilm dynamics, with implications for managing biofilm-related infections and industrial biofouling. [ABSTRACT FROM AUTHOR]

The cuticle covering the outer surface of an eggshell functions as both a physical and chemical barrier against invading microorganisms. Contamination of eggs by microbial pathogens progresses in four stages: bacterial attachment to the egg surface, penetration through the cuticle and eggshell, multiplication within the underlying membranes, and the final stage of contaminating the egg contents. Therefore, it is important to study bacterial count at the first point of contact, i.e., on the surface of the eggs. In this study, we have evaluated the impact of differences in cuticle quality (due to egg washing and hen age) on bacterial load. We compared bacterial adherence on the eggshell surface of white eggs which were either washed (graded) or unwashed (ungraded), collected from Lohmann laying hens of different ages: early (24–28 weeks), mid-lay (44–48 weeks), and late (66–70 weeks). We aimed to determine the impact of hen age and egg washing on differences in cuticle quality and bacterial adherence. Our results indicate that hen age (up to 70 weeks) and commercial egg washing do not significantly impact bacterial adherence on eggshell surfaces. We have developed a novel method using green fluorescent protein (GFP)-expressing Salmonella typhimurium to estimate adherence of bacteria to the eggshell surface, with independent measurement of autofluorescence to quantitate cuticle deposition. S. typhimurium were localized, adhering to cracks visible on the outer cuticle in ungraded eggs, indicating that egg-associated pathogens usually enter the egg interior either through respiratory pores in eggshells or through shell micro-cracks. The results of this study can be utilized to optimize innovative methods for predictive microbiology in order to achieve egg safety. [ABSTRACT FROM AUTHOR]

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The interaction between bacteria and the intestinal mucus is crucial during the early pathogenesis of many enteric diseases in mammals. A critical step in this process employed by both commensal and pathogenic bacteria focuses on the breakdown of the protective layer presented by the intestinal mucus by mucolytic enzymes. C. perfringens type G, the causative agent of necrotic enteritis in broilers, produces two glycosyl hydrolase family 18 chitinases, ChiA and ChiB, which display distinct substrate preferences. Whereas ChiB preferentially processes linear substrates such as chitin, ChiA prefers larger and more branched substrates, such as carbohydrates presented by the chicken intestinal mucus. Here, we show via crystal structures of ChiA and ChiB in the apo and ligand-bound forms that the two enzymes display structural features that explain their substrate preferences providing a structural blueprint for further interrogation of their function and inhibition. This research focusses on the roles of ChiA and ChiB in bacterial proliferation and mucosal attachment, two processes leading to colonization and invasion of the gut. ChiA and ChiB, either supplemented or produced by the bacteria, led to a significant increase in C. perfringens growth. In addition to nutrient acquisition, the importance of chitinases in bacterial attachment to the mucus layer was shown using an in vitro binding assay of C. perfringens to chicken intestinal mucus. Both an in vivo colonization trial and a necrotic enteritis trial were conducted, demonstrating that a ChiA chitinase mutant strain was less capable to colonize the intestine and was hampered in its disease-causing ability as compared to the wild-type strain. Our findings reveal that the pathogen-specific chitinases produced by C. perfringens type G strains play a fundamental role during colonization, suggesting their potential as vaccine targets. Author summary: The intestinal mucus layer protects the intestinal mucosa from invasion by pathogenic bacteria. However, it is precisely this first line of protection that invading bacteria aim to break down en route to colonizing the gut. Clostridium perfringens type G, the causative agent of necrotic enteritis in broiler chickens, an enteric disease characterised by ulcers in the small intestine, employs such a pathogenic strategy. In addition to the well-studied NetB toxin, this bacterium can produce other pathogen-specific enzymes, some of which could play a central role during disease development. Two of these enzymes were identified as putative chitinases, ChiA and ChiB. We have demonstrated that ChiA and ChiB are indeed functional enzymes that can hydrolyze a wide variety of substrates, with ChiA preferring mucus as a substrate. Mucus breakdown by the chitinases induces both bacterial proliferation and attachment to mucus, two processes that are crucial in the first steps of the pathogenesis. In addition, we have shown that a ChiA mutant strain does not efficiently colonize the intestine and causes less severe lesions as compared to the wild-type strain. Thus, we conclude that chitinases provide an advantage to the pathogen to facilitate colonization of the intestine of broiler chickens. [ABSTRACT FROM AUTHOR]

Enterotoxigenic Escherichia coli (ETEC) cause hundreds of millions of cases of infectious diarrhea annually, predominantly in children from low-middle income regions. Notably, in children, as well as volunteers challenged with ETEC, diarrheal severity is significantly increased in blood group A (bgA) individuals. EtpA, is a secreted glycoprotein adhesin that functions as a blood group A lectin to promote critical interactions between ETEC and blood group A glycans on intestinal epithelia for effective bacterial adhesion and toxin delivery. EtpA is highly immunogenic resulting in robust antibody responses following natural infection and experimental challenge of volunteers with ETEC. To understand how EtpA directs ETEC-blood group A interactions and stimulates adaptive immunity, we mutated EtpA, mapped its glycosylation by mass-spectrometry (MS), isolated polyclonal (pAbs) and monoclonal antibodies (mAbs) from vaccinated mice and ETEC-infected volunteers, and determined structures of antibody-EtpA complexes by cryo-electron microscopy. Both bgA and mAbs that inhibited EtpA-bgA interactions and ETEC adhesion, bound to the C-terminal repeat domain highlighting this region as crucial for ETEC pathogen-host interaction. MS analysis uncovered extensive and heterogeneous N-linked glycosylation of EtpA and cryo-EM structures revealed that mAbs directly engage these unique glycan containing epitopes. Finally, electron microscopy-based polyclonal epitope mapping revealed antibodies targeting numerous distinct epitopes on N and C-terminal domains, suggesting that EtpA vaccination generates responses against neutralizing and decoy regions of the molecule. Collectively, we anticipate that these data will inform our general understanding of pathogen-host glycan interactions and adaptive immunity relevant to rational vaccine subunit design. Author summary: Enterotoxigenic E. coli (ETEC), a leading cause of diarrhea disproportionately affecting young children in low-income regions, are a priority for vaccine development. Individuals possessing A blood-type are more susceptible to severe cholera-like disease. EtpA, a secreted, immunogenic, blood group A binding protein, is a current vaccine target antigen. Here, we determined the structure of EtpA in complex with protective as well as non-protective monoclonal antibodies targeting two different domains of the protein, pinpointing key regions involved in blood-group A antigen recognition and uncovering the mechanism of antibody-based protection. In addition, we show through mass-spectrometry that EtpA is extensively and heterogeneously glycosylated at surface-exposed asparagine residues by a promiscuous and low-fidelity glycosyltransferase, EtpC, and that this unique form of bacterial glycosylation is critical for to development of protective immune responses. Lastly, polyclonal antibodies from vaccinated mice as well as monoclonal antibodies obtained from ETEC-infected volunteers revealed that the highly antigenic surface of EtpA exhibits both protective and non-protective epitopes. These results greatly expand our understanding of ETEC pathogenesis, and the immune responses elicited by these common infections, providing valuable information to aid in the rational design and testing of subunit vaccines. [ABSTRACT FROM AUTHOR]

Bacterial pathogens are responsible for a variety of human diseases, necessitating their prompt detection for effective diagnosis and treatment of infectious diseases. Over recent years, electrochemical methods have gained significant attention owing to their exceptional sensitivity and rapidity. This review outlines the current landscape of electrochemical biosensors employed in clinical diagnostics for the detection of bacterial pathogens. We categorize these biosensors into four types: amperometry, potentiometry, electrochemical impedance spectroscopy, and conductometry, targeting various bacterial components, including toxins, virulence factors, metabolic activity, and events related to bacterial adhesion and invasion. We discuss the merits and challenges associated with electrochemical methods, underscoring their rapid response, high sensitivity, and specificity, while acknowledging the necessity for skilled operators and potential interference from biological and environmental factors. Furthermore, we examine future prospects and potential applications of electrochemical biosensors in clinical diagnostics. While electrochemical biosensors offer a promising avenue for detecting bacterial pathogens, further research in optimizing the robustness and surmounting the challenges hindering their seamless integration into clinical practice is imperative. [ABSTRACT FROM AUTHOR]

Porcine circovirus type 2 (PCV2) often causes disease through coinfection with other bacterial pathogens, including Glaesserella parasuis (G. parasuis), which causes high morbidity and mortality, but the role played by PCV2 and bacterial and host factors contributing to this process have not been defined. Bacterial attachment is assumed to occur via specific receptor-ligand interactions between adhesins on the bacterial cell and host proteins adsorbed to the implant surface. Mass spectrometry (MS) analysis of PCV2-infected swine tracheal epithelial cells (STEC) revealed that the expression of Extracellular matrix protein (ECM) Fibronectin (Fn) increased significantly on the infected cells surface. Importantly, efficient G. parasuis serotype 4 (GPS4) adherence to STECs was imparted by interactions with Fn. Furthermore, abrogation of adherence was gained by genetic knockout of Fn, Fn and Integrin β1 antibody blocking. Fn is frequently exploited as a receptor for bacterial pathogens. To explore the GPS4 adhesin that interacts with Fn, recombinant Fn N-terminal type I and type II domains were incubated with GPS4, and the interacting proteins were pulled down for MS analysis. Here, we show that rare lipoprotein A (RlpA) directly interacts with host Fibronectin mediating GPS4 adhesion. Finally, we found that PCV2-induced Fibronectin expression and adherence of GPS4 were prevented significantly by TGF-β signaling pathway inhibitor SB431542. Our data suggest the RlpA-Fn interaction to be a potentially promising novel therapeutic target to combat PCV2 and GPS4 coinfection. Author summary: Porcine circovirus type 2 and Glaesserella parasuis (G. parasuis) are common upper respiratory tract pathogens in pigs, which co-infection causes high morbidity and mortality. However, the underlying mechanisms of host-pathogen interactions during PCV2 and GPS4 co-infection remain unclear. Here, we focused on the interaction of PCV2-infected cells and GPS4, which revealed that swine tracheal epithelial cells (STEC) infected with PCV2 exhibit the increased expression of Fn. Notably, PCV2 infection-induced Fn interacting with RlpA was shown to impart efficient adherence to GPS4. Fn and Integrin β1 antibody blocking and SB431542 counteracted PCV2 infection-induced GPS4 colonization. These studies reveal a novel receptor-ligand interaction that enhances colonization of the tracheal by GPS4 following PCV2 infection and highlights the importance of Fn-RlpA in host-pathogen interactions. In addition, RlpA was discovered for the first time as an adhesin of GPS4, which provided a new idea for the development of G. parasuis vaccines. [ABSTRACT FROM AUTHOR]

In this study, zeolitic imidazolate framework 8 (ZIF-8) was coated on porous Ti6Al4V scaffolds, either bare or previously modified using hydroxyapatite (HA) or HA and gelatin (HAgel), via a growing single-step method in aqueous media using two contact times at 6 h and 24 h. The coated scaffolds termed ZIF-8@Ti, ZIF-8@ HA/Ti, and ZIF-8@HAgel/Ti were characterized via scanning electron microscopy (SEM), powder X-ray diffraction (PXRD), attenuated total reflectance-Fourier transform infrared (ATR-FTIR), and molecular plasma-atomic emission spectroscopy (MP-AES). In order to assess the cell proliferation rate, the cytocompatibility of the scaffolds was evaluated in primary osteoblasts (hOBs) using alamarBlue assay, while the osteoconductivity was analyzed in hOBs using a real-time approach, evaluating the expression of secreted phosphoprotein 1 (SPP1). Osteopontin, which is the protein encoded by this gene, represents the major non-collagenous bone protein that binds tightly to HA. The scaffolds were shown to be non-cytotoxic based on hOB proliferation at all time points of analysis (24 h and 72 h). In hOB cultures, the scaffolds induced the upregulation of SPP1 with different fold changes. Some selected scaffolds were assayed in vitro for their antibacterial potential against Staphylococcus epidermidis; the scaffolds coated with ZIF-8 crystals, regardless of the presence of HA and gelatin, strongly inhibited bacterial adhesion to the materials and reduced bacterial proliferation in the culture medium, demonstrating the suitable release of ZIF-8 in a bioactive form. These experiments suggest that the innovative scaffolds, tested herein, provide a good microenvironment for hOB adhesion, viability, and osteoconduction with effective prevention of S. epidermidis adhesion. [ABSTRACT FROM AUTHOR]

Background: Type VI secretion system (T6SS) is widely present in Gram-negative bacteria and directly mediates antagonistic prokaryote interactions. PAAR (proline-alanine-alanine-arginine repeats) proteins have been proven essential for T6SS-mediated secretion and target cell killing. Although PAAR proteins are commonly found in A. baumannii, their biological functions are not fully disclosed yet. In this study, we investigated the functions of a PAAR protein termed TagP (T6SS-associated-gene PAAR), encoded by the gene ACX60_RS09070 outside the core T6SS locus of A. baumannii strain ATCC 17978. Methods: In this study, tagP null and complement A. baumannii ATCC 17978 strains were constructed. The influence of TagP on T6SS function was investigated through Hcp detection and bacterial competition assay; the influence on environmental fitness was studied through in vitro growth, biofilm formation assay, surface motility assay, survivability in various simulated environmental conditions; the influence on pathogenicity was explored through cell adhesion and invasion assays, intramacrophage survival assay, serum survival assay, and G. melonella Killing assays. Quantitative transcriptomic and proteomic analyses were utilized to observe the global impact of TagP on bacterial status. Results: Compared with the wildtype strain, the tagP null mutant was impaired in several tested phenotypes such as surface motility, biofilm formation, tolerance to adverse environments, adherence to eukaryotic cells, endurance to serum complement killing, and virulence to Galleria melonella. Notably, although RNASeq and proteomics analysis revealed that many genes were significantly downregulated in the tagP null mutant compared to the wildtype strain, there is no significant difference in their antagonistic abilities. We also found that Histonelike nucleoid structuring protein (H-NS) was significantly upregulated in the tagP null mutant at both mRNA and protein levels. Conclusions: This study enriches our understanding of the biofunction of PAAR proteins in A. baumannii. The results indicates that TagP involved in a unique modulation of fitness and virulence control in A. baumannii, it is more than a classic PAAR protein involved in T6SS, while how TagP play roles in the fitness and virulence of A. baumannii needs further investigation to clarify. [ABSTRACT FROM AUTHOR]

Microbial cells serve as efficient and environmentally friendly biocatalysts, but their stability and reusability in practical applications must often be improved through immobilization. Acinetobacter sp. Tol 5 shows high adhesiveness to materials due to its large cell surface protein AtaA, which consists of 3630 amino acids (aa). Previously, we developed a method for immobilizing bacteria using AtaA. Herein, we investigated the cell immobilization ability of in-frame deletion (IFD) mutants of AtaA with different sizes in Tol 5. Mini-AtaA, which consists of 775 aa and is functional in Escherichia coli, was produced and present on the cell surface; however, mini-AtaA showed no immobilization ability in Tol 5. A cell immobilization assay was performed with cells expressing 16 IFD mutants of AtaA with different sizes, revealing that a length of at least 1417 aa was required for the sufficient immobilization of Tol 5 cells; thus, the minimum length needed to achieve the adhesive function of AtaA varies among bacterial species. The constructed mutant library of AtaA ranging from 3630 to 775 aa will allow researchers to quickly and easily explore the optimal size of AtaA, even for bacteria newly introduced to AtaA. [ABSTRACT FROM AUTHOR]

Laser texturing seems to be a promising technique for reducing bacterial adhesion on titanium implant surfaces. This work aims to demonstrate the possibility of obtaining a functionally orientated surface of titanium implant elements with a specific architecture with specific bacteriological and photocatalytic properties. Femtosecond laser-generated surface structures, such as laser-induced periodic surface structures (LIPSS, wrinkles), grooves, and spikes on titanium, have been characterised by XRD, Raman spectroscopy, and scanning electron microscopy (SEM). The photocatalytic activity of the titanium surfaces produced was tested based on the degradation effect of methylene blue (MB). The correlation between the photocatalytic activity of TiO2 coatings and their morphology and structure has been analysed. Features related to the size, shape, and distribution of the roughness patterns were found to influence the adhesion of the bacterial strain on different surfaces. On the laser-structurised surface, the adhesion of Escherichia coli bacteria were reduced by 80% compared to an untreated reference surface. [ABSTRACT FROM AUTHOR]

The chronic airway infections with Pseudomonas aeruginosa are the major comorbidity in people with cystic fibrosis (CF). Within CF lungs, P. aeruginosa persists in the conducting airways together with human mucins as the most abundant structural component of its microenvironment. We investigated the adhesion of 41 serial CF airway P. aeruginosa isolates to airway mucin preparations from CF sputa. Mucins and bacteria were retrieved from five modulator-naïve patients with advanced CF lung disease. The P. aeruginosa isolates from CF airways and non-CF reference strains showed a strain-specific signature in their adhesion to ovine, porcine and bovine submaxillary mucins and CF airway mucins ranging from no or low to moderate and strong binding. Serial CF clonal isolates and colony morphotypes from the same sputum sample were as heterogeneous in their affinity to mucin as representatives of other clones thus making 'mucin binding' one of the most variable intraclonal phenotypic traits of P. aeruginosa known to date. Most P. aeruginosa CF airway isolates did not adhere more strongly to CF airway mucins than to plastic surfaces. The strong binders, however, exhibited a strain-specific affinity gradient to O-glycans, CF airway and mammalian submaxillary mucins. [ABSTRACT FROM AUTHOR]

Antimicrobial potential of bioactive glass (BAG) makes it promising for implant applications, specifically overcoming the toxicity concerns associated with traditional antibacterial nanoparticles. The 58S composition of BAG (with high Ca and absence of Na) has been known to exhibit excellent bioactivity and antibacterial behaviour, but the mechanisms behind have not been investigated in detail. In this pioneering study, we are using Atomic Force Microscopy (AFM) to gain insights into 58S BAG's adhesive interactions with planktonic cells of both gram-positive (Staphylococcus aureus) and gram-negative (Escherichia coli) bacteria; along with the impact of crystallinity on antibacterial properties. We have recorded greater bacterial inhibition by amorphous BAG compared to semi-crystalline glass–ceramics and stronger effect against gram-negative bacteria via conventional long-term antibacterial tests. AFM force distance curves has illustrated substantial bonding between bacteria and BAG within the initial one second (observed at a gap of 250 ms) of contact, with multiple binding events. Further, stronger adhesion of BAG with E.coli (~ 6 nN) compared to S. aureus (~ 3 nN) has been found which can be attributed to more adhesive nano-domains (size effect) distributed uniformly on E.coli surface. This study has revealed direct evidence of impact of contact time and 58S BAG's crystalline phase on bacterial adhesion and antimicrobial behaviour. Current study has successfully demonstrated the mode and mechanisms of initial bacterial adhesion with 58S BAG. The outcome can pave the way towards improving the designing of implant surfaces for a range of biomedical applications. [ABSTRACT FROM AUTHOR]

Background: Chlorhexidine gluconate (CHG) (0.05%) has recently been suggested as both a dip for the hydrophilic surface and an irrigation solution in the setting of penile prosthesis (PP) surgery. Aim: The study sought to compare the antimicrobial efficacy of 0.05% CHG with vancomycin and gentamicin (VG) antibiotics as dip and/or irrigation solutions in the setting of a hydrophilic PP surface in vitro. Methods: Sterile PPs with a hydrophilic coating were obtained. A series of experiments were performed to evaluate the efficacy of normal saline (NS), 0.05% CHG, or VG as dip and/or irrigation solutions to reduce methicillin-sensitive Staphylococcus aureus adhesion to PP surfaces. The 8-mm discs from PPs were incubated in 105 colony-forming units/mL of methicillin-sensitive S aureus for 48 hours, plated, and counted. Disc-diffusion tests were conducted by suspending 6-mm discs for 2 minutes in NS, 0.05% CHG, or VG, then placing them coated side down onto plates streaked with the following organisms: methicillin-sensitive S aureus, S epidermidis, Enterococcus, and Escherichia coli. After 24 hours of growth, zones of inhibition were measured. Outcomes: We found average bacterial counts (colony-forming units/mL) and zones of inhibition (mm) following a series of treatment protocols of PP discs. Results: PP discs dipped in VG reduced bacterial adhesion to the implant surface >0.05% CHG (~5.5 log vs ~1.5 log; P < .01). Discs irrigated with either 0.05% CHG or NS removed all dip solution adsorbed to the hydrophilic surface, allowing bacterial growth. VG irrigation adsorbed to the hydrophilic surface even after 0.05% CHG or NS dips, reducing bacterial adherence (~3 log). Dipping and irrigating discs with VG was most effective in reducing adherent bacteria (~5.5 log) and was the only irrigation that showed antimicrobial activity. Clinical Translation: VG, when used both as a prophylactic dip and as an intraoperative irrigation solution for hydrophilic penile implant surfaces, has improved efficacy to 0.05% CHG and NS. Strengths and Limitations: This is the first study to compare the use of VG, 0.05% CHG, and NS as prophylactic dips and intraoperative irrigations for hydrophilic penile implant surfaces. Limitations include the use of in vitro studies, which serve as a proxy for in vivo practices and may not be entirely accurate nor translatable clinically. Conclusion: We demonstrated the superior efficacy of VG as a combined dip and irrigation solution for hydrophilic penile implant surfaces compared with 0.05% CHG. [ABSTRACT FROM AUTHOR]

The terrestrial biota is a rich source of biologically active substances whose anti-biofilm potential is not studied enough. The aim of this review is to outline a variety of terrestrial sources of antimicrobial agents with the ability to inhibit different stages of biofilm development, expecting to give some ideas for their utilization in improved anti-biofilm treatments. It provides an update for the last 5 years on anti-biofilm plant products and derivatives, essential oils, antimicrobial peptides, biosurfactants, etc., that are promising candidates for providing novel alternative approaches to combating multidrug-resistant biofilm-associated infections. Based on the reduction in bacterial adhesion to material and cell surfaces, the anti-adhesion strategy appears interesting for the prevention of bacterial attachment in combating a broad range of mono- and multispecies bacterial biofilms. So far, few studies have been carried out in this direction. Anti-biofilm coatings made by or containing biologically active products from terrestrial biota have scarcely been studied although they are of significant interest for a reduction in infections associated with medical devices. Combination therapy with commercial antibiotics and natural products is accepted now as a promising base for future advances in anti-biofilm treatment. In vivo testing and clinical trials are necessary for clinical application. [ABSTRACT FROM AUTHOR]

Endocarditis is a severe infection affecting the heart's inner layer, the endocardium. Its pathophysiology may involve heart valve damage, bacteria adhesion and biofilm formation, potentially leading to fatal complications. Bacteria from various sources, including from endodontic diseases and its treatments may enter the bloodstream provoking this condition. This systematic review aimed to explore the influence of endodontic factors on endocarditis. Searches across PubMed, Embase, Cochrane Library and manual sources yielded 14 relevant articles from 1562 screened studies. Assessment platforms from JBI Critical Appraisal Tools evaluated studies biases. Findings mainly focused on transient bacteraemia as a key indicator of risk correlating bacterial virulence and counts with endocarditis development. Worryingly, multi-species bacteraemia post-endodontic treatment was noted including the genera Enterococcus, Parvimonas, Streptococcus and Staphylococcus. Conclusive validation of the incidence and association between endodontic patients and endocarditis was limited due to a lack of robust longitudinal investigations, such as randomized controlled trials. This emphasizes the need for further research with well-designed methodologies to provide a full understanding of the causative bacterial population and its pathological mechanisms. A current guideline (2023 European Society of Cardiology) was developed to support healthcare professionals in diagnosing and managing infective endocarditis; this 2023 version is introducing a new diagnostic algorithm to aid in patient classification aiming to improve outcomes for this challenging disease. The study was a priori registered on PROSPERO (CRD42023407736). [ABSTRACT FROM AUTHOR]

Polymer-based textiles have a major impact on human well-being, as they provide the desired functional protection and aesthetic comfort when worn. However, natural and synthetic polymer-based textiles can also pose serious health risks, as they are surfaces that allow the adhesion of various bacteria, including pathogenic bacteria. To minimise these problems, antibacterial chemical treatments are generally applicable in the case of polymer-based textiles. However, to avoid the use of potentially toxic chemicals, sustainable approaches require the customised design of non-adhesive polymer-based textiles, considering their chemical, physicochemical, constructional, and textural properties. Before designing, several articles are required to gain sufficient knowledge of the described object. Despite the urgent need to combat bacteria (on polymer-based textiles), which pose a serious global health risk, only a few review articles have been published that address bacterial adhesion in the context of superhydrophobic and antibacterial textile materials, while only one review article holistically addresses the textile factors and their influence on this phenomenon. The aim of this review article is to expand the insufficient knowledge about bacterial adhesion to polymer-based textiles on the basis of theoretical findings and real examples through a high degree of structuring, simplification, holistic consideration, and visualization. Therefore, this review provides an insight into the mechanisms involved in bacterial adhesion and a comprehensive overview of the influence of different textile factors, such as chemical composition, hydrophilicity/hydrophobicity, surface charge, surface free energy, roughness, and porosity, on bacterial adhesion. To emphasise the importance of the synergistic effect of the combined textile factors, examples of the influence of hydrophilicity/hydrophobicity in combination with surface charge, surface roughness, and porosity are discussed. From the review, it can be concluded that the combination of hydrophilicity/hydrophobicity and the surface charge of textile fibres and bacteria is crucial for bacterial adhesion, with roughness and porosity being the most important factors among the constructive and textural properties of polymer-based textiles. [ABSTRACT FROM AUTHOR]

'Active targeting' refers to modifying a nanocarrier's surface with targeting ligands. This study introduced an efficient approach for immobilizing imidazole-based drugs onto the metallated-porphyrin complex within the porphysome nanocarrier. To enhance cellular and bacterial uptake, a Ni-porphyrin with a fatty acid tail was synthesized and placed in the bilayer center of DPPC, facilitating receptor-mediated endocytosis. The Ni-porphyrin in the head group of the Ni-porphyrin-tail was placed superficially in the polar region of the membrane. Spherical unilamellar vesicle formation (DPPC: Ni-porphyrin-tail 4:1 mole ratio), as metallo-porphysome, was achieved through supramolecular self-assembly in an aqueous buffer. These vesicles exhibited a diameter of 279 ± 7 nm and a zeta potential of −15.3 ± 2.5 mV, showcasing their unique cytocompatibility. Nitroimidazole was decorated on the surface of metallo-porphysomes and pistachio green hull extract (PGHE) was loaded into the carrier for synergistic activity against (E. coli) and (S. aureus) bacteria strains. The physicochemical properties of Nitroimidazole-porphysome-PGHE, including size, zeta potential, morphology, loading efficiency, and release profile under various pH and temperature conditions in simulated gastrointestinal fluids were characterized. This combination therapy prevented bacterial cell attachment and biofilm formation in Caco-2 cells, as colon epithelial cells. The remarkable benefit of this system is that it does not affect cell viability even at 0.5 mg/ml. This study demonstrates the potential of a new co-delivery system using biocompatible metallo-porphysomes to decrease bacterial infections. [ABSTRACT FROM AUTHOR]

Infections pose a challenge for the fast growing aquaculture sector. Glycosphingolipids are cell membrane components that pathogens utilize for attachment to the host to initiate infection. Here, we characterized rainbow trout glycosphingolipids from five mucosal tissues using mass spectrometry and nuclear magnetic resonance and investigated binding of radiolabeled Aeromonas salmonicida to the glycosphingolipids on thin-layer chromatograms. 12 neutral and 14 acidic glycosphingolipids were identified. The glycosphingolipids isolated from the stomach and intestine were mainly neutral, whereas glycosphingolipids isolated from the skin, gills and pyloric caeca were largely acidic. Many of the acidic structures were poly-sialylated with shorter glycan structures in the skin compared to the other tissues. The sialic acids found were Neu5Ac and Neu5Gc. Most of the glycosphingolipids had isoglobo and ganglio core chains, or a combination of these. The epitopes on the rainbow trout glycosphingolipid glycans differed between epithelial sites leading to differences in pathogen binding. A major terminal epitope was fucose, that occurred attached to GalNAc in a α1-3 linkage but also in the form of HexNAc-(Fuc-)HexNAc-R. A. salmonicida were shown to bind to neutral glycosphingolipids from the gill and intestine. This study is the first to do a comprehensive investigation of the rainbow trout glycosphingolipids and analyze binding of A. salmonicida to glycosphingolipids. The structural information paves the way for identification of ways of interfering in pathogen colonization processes to protect against infections in aquaculture and contributes towards understanding A. salmonicida infection mechanisms. [ABSTRACT FROM AUTHOR]

Polymeric elastomers are widely utilized in implantable biomedical devices. Nevertheless, the implantation of these elastomers can provoke a robust foreign body response (FBR), leading to the rejection of foreign implants and consequently reducing their effectiveness in vivo. Building effective anti-FBR coatings on those implants remains challenging. Herein, we introduce a coating-free elastomer with superior immunocompatibility. A super-hydrophilic anti-fouling zwitterionic layer can be generated in situ on the surface of the elastomer through a simple chemical trigger. This elastomer can repel the adsorption of proteins, as well as the adhesion of cells, platelets, and diverse microbes. The elastomer elicited negligible inflammatory responses after subcutaneous implantation in rodents for 2 weeks. No apparent fibrotic capsule formation was observed surrounding the elastomer after 6 months in rodents. Continuous subcutaneous insulin infusion (CSII) catheters constructed from the elastomer demonstrated prolonged longevity and performance compared to commercial catheters, indicating its great potential for enhancing and extending the performance of various implantable biomedical devices by effectively attenuating local immune responses. The foreign body response remains a significant challenge for implants. Complicated coating procedures are usually needed to construct anti-fibrotic coatings on implantable elastomers. Herein, a coating-free elastomer with superior immunocompatibility was achieved using a zwitterionic monomer derivative. A pure zwitterionic layer can be generated on the elastomer surface through a simple chemical trigger. This elastomer significantly reduces protein adsorption, cell and bacterial adhesion, and platelet activation, leading to minimal fibrotic capsule formation even after six months of subcutaneous implantation in rodents. CSII catheters constructed from the PQCBE-H elastomer demonstrated prolonged longevity and performance compared to commercial catheters, highlighting the significant potential of PQCBE-H elastomers for enhancing and extending the performance of various implantable biomedical devices. [Display omitted] [ABSTRACT FROM AUTHOR]

Atomic force microscopy was utilized to estimate the adhesion strengths to silicon nitride as well as the cellular elasticities of pathogenic Listeria monocytogenes EGDe cells cultured in media adjusted to five different pH conditions of growth (5, 6, 7, 8, and 9) under water with 0.0027 fixed ionic strength. Particularly, the role of adhesion on the bacterial elastic properties was investigated. The nonadhesive Hertz model of contact mechanics was used to extract Young's moduli of elasticity of bacterial cells from the approach force-indentation data. Additionally, the adhesive models of contact mechanics: Johnson–Kendall–Roberts (JKR) and Derjaguin–Muller–Toporov (DMT) were used to estimate Young's moduli of elasticity of bacterial cells from the retraction force-indentation data. Our results indicated that adhesion to silicon nitride was the highest for cells cultured at a pH of 7. Similarly, bacterial cells cultured at pH 7 were characterized by the highest Young's moduli of elasticities compared to the lower or higher pH conditions of growth. Young's moduli of elasticities estimated from the Hertz model were stiffer than those estimated using JKR or DMT models. As the adhesion between bacterial cells and indenters increased, the difference between the Hertz model and JKR or DMT models estimates of Young's moduli of elasticity increased as well. Contradicting the current norm of using the Hertz model to quantify bacterial elasticity in the literature, our results highlight the extreme importance of utilizing contact mechanics models with adhesion components in them such as the JKR and DMT models to estimate bacterial elasticity. [ABSTRACT FROM AUTHOR]