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Aim. This study investigates the demographic distribution, antibiotic resistance profiles, and molecular characteristics of Staphylococcus aureus infections. Methods. The study was carried out in 141 patients, 60.4% male, in patients from Chania and Heraklion, Crete. Results. The highest infection prevalence observed in the older adults (≥65 years) age group. The predominant infection types were skin lesions (39.72) and respiratory tract infection (22.7%). Antibiotic resistance testing revealed that 57.44% of strains were MRSA, with high resistance to Tetracycline, Ciprofloxacin, Kanamycine Erythromycin and Clindamycin. Molecular analysis showed 19.14% of strains were Pvl-positive, highlighting the presence of both MRSA and MSSA strains with Pvl genes. Conclusions. The study underscores the need for continuous surveillance and targeted infection control strategies to manage the spread of MRSA, particularly in vulnerable populations. [ABSTRACT FROM AUTHOR]

Oleate hydratase (OhyA), a flavoenzyme that catalyzes the hydration of unsaturated fatty acids, has been identified in various Bacillales organisms, including those in the Listeria, Lysinibacillus, Paenibacillus, and Staphylococcus genera. In this study, we combine structural biology with molecular and phylogenetic analyses to investigate the evolutionary dynamics of the OhyA protein family within the Bacillales order. Our evolutionary analysis reveals two distinct OhyA clades (clade I and clade II) within Bacillales that, while sharing catalytic function, exhibit significant genomic and structural differences. Our findings suggest that these OhyA clades originated from independent evolutionary processes through convergent evolution rather than gene duplication. We also show that the evolutionary divergence in OhyA is likely due to intrinsic sequence variations rather than being strictly linked to functional domain changes. Furthermore, within the Staphylococcus genus, we observed that the evolution of the ohyA gene aligns with the species tree, supporting a common ancestral origin. This study enhances our understanding of the impact of evolutionary history on the structure and function of OhyA across the Bacillales order.

Stainless steel 316L (SS316L) is widely used in fracture management devices. However, SS316L does not offer any bacterial infection resistance and can cause metal-ion sensitivity due to Ni-ions’ presence. 17-4PH can emerge as a promising substitute due to the intrinsic antibacterial properties of copper, a 75% reduction in nickel content, and superior mechanical properties. SS316L and 17–4 PH were manufactured using laser-directed energy deposition (LDED). 17-4PH specimens surpassed the compressive strength of SS316L by over 150%. A static magnetic field was generated in 17–4 PH specimens to understand in vitro bone cell–material interactions. In vitro human fetal osteoblast cell culture and bacterial inhibition study using Staphylococcus aureus and Pseudomonas aeruginosa were carried out on these specimens with SS316L as control and as-processed and magnetised 17–4 PH as treatments. Results demonstrated that magnetised 17–4 PH exhibited 25% enhancement in hFOB proliferation and 70% reduction in bacterial colonisation compared to SS316L.

Abstract Background There is an urgent clinical need for developing novel immunoprophylaxis and immunotherapy strategies against Staphylococcus aureus (S. aureus). In our previous work, immunization with a tetra-branched multiple antigenic peptide, named MAP2-3 that mimics lipoteichoic acid, a cell wall component of S. aureus, successfully induced a humoral immune response and protected BALB/c mice against S. aureus systemic infection. In this study, we further investigated whether vaccination with MAP2-3 can elicit immunologic memory. Methods BALB/c mice were immunized with MAP2-3 five times. After one month of the last vaccination, mice were challenged with heat-killed S. aureus via intraperitoneal injection. After a 7-day inoculation, the percentage of plasma cells, memory B cells, effector memory T cells, and follicular helper T cells were detected by flow cytometry. The levels of IL-6, IL-21, IL-2, and IFN-γ were measured by real-time PCR and ELISA. Flow cytometry results were compared by using one-way ANOVA or Mann-Whitney test, real-time PCR results were compared by using one-way ANOVA, and ELISA results were compared by using one-way ANOVA or student’s t-test. Results The percentage of plasma cells and memory B cells in the spleen and bone marrow from the MAP2-3 immunized mice was significantly higher than that from the control mice. The percentage of effector memory T cells in spleens and lymphoid nodes as well as follicular helper T cells in spleens from the MAP2-3 immunized mice were also higher. Moreover, the levels of IL-6 and IL-21, two critical cytokines for the development of memory B cells, were significantly higher in the isolated splenocytes from immunized mice after lipoteichoic acid stimulation. Conclusions Immunization with MAP2-3 can efficiently induce memory B cells and memory T cells.

Background: There is an urgent clinical need for developing novel immunoprophylaxis and immunotherapy strategies against Staphylococcus aureus (S. aureus). In our previous work, immunization with a tetra-branched multiple antigenic peptide, named MAP2-3 that mimics lipoteichoic acid, a cell wall component of S. aureus, successfully induced a humoral immune response and protected BALB/c mice against S. aureus systemic infection. In this study, we further investigated whether vaccination with MAP2-3 can elicit immunologic memory. Methods: BALB/c mice were immunized with MAP2-3 five times. After one month of the last vaccination, mice were challenged with heat-killed S. aureus via intraperitoneal injection. After a 7-day inoculation, the percentage of plasma cells, memory B cells, effector memory T cells, and follicular helper T cells were detected by flow cytometry. The levels of IL-6, IL-21, IL-2, and IFN-γ were measured by real-time PCR and ELISA. Flow cytometry results were compared by using one-way ANOVA or Mann-Whitney test, real-time PCR results were compared by using one-way ANOVA, and ELISA results were compared by using one-way ANOVA or student's t-test. Results: The percentage of plasma cells and memory B cells in the spleen and bone marrow from the MAP2-3 immunized mice was significantly higher than that from the control mice. The percentage of effector memory T cells in spleens and lymphoid nodes as well as follicular helper T cells in spleens from the MAP2-3 immunized mice were also higher. Moreover, the levels of IL-6 and IL-21, two critical cytokines for the development of memory B cells, were significantly higher in the isolated splenocytes from immunized mice after lipoteichoic acid stimulation. Conclusions: Immunization with MAP2-3 can efficiently induce memory B cells and memory T cells. [ABSTRACT FROM AUTHOR]

Introduction: During the COVID-19 Delta variant surge, the CLAIRE crosssectional study sampled saliva from 120 hospitalized patients, 116 of whom had a positive COVID-19 PCR test. Patients received antibiotics upon admission due to possible secondary bacterial infections, with patients at risk of sepsis receiving broad-spectrum antibiotics (BSA). Methods: The saliva samples were analyzed with shotgun DNA metagenomics and respiratory RNA virome sequencing. Medical records for the period of hospitalization were obtained for all patients. Once hospitalization outcomes were known, patients were classified based on their COVID-19 disease severity and the antibiotics they received. Results: Our study reveals that BSA regimens differentially impacted the human salivary microbiome and disease progression. 12 patients died and all of them received BSA. Significant associations were found between the composition of the COVID-19 saliva microbiome and BSA use, between SARS-CoV-2 genome coverage and severity of disease.We also found significant associations between the non-bacterial microbiome and severity of disease, with Candida albicans detected most frequently in critical patients. For patients who did not receive BSA before saliva sampling, our study suggests Staphylococcus aureus as a potential risk factor for sepsis. Discussion: Our results indicate that the course of the infection may be explained by both monitoring antibiotic treatment and profiling a patient’s salivary microbiome, establishing a compelling link between microbiome and the specific antibiotic type and timing of treatment. This approach can aid with emergency room triage and inpatient management but also requires a better understanding of and access to narrow-spectrum agents that target pathogenic bacteria. [ABSTRACT FROM AUTHOR]

Polyethersulfone (PES) membranes are prevalent in the field of water treatment owing to their exceptional separation efficiency, robust mechanical properties, and resistance to chemical degradation. Nevertheless, these membranes are prone to fouling, resulting in a decrease in both flux and ultrafiltration efficiency. In the present study, PES membranes are blended with poly (3‐Sulfopropyl Methacrylate) (PSPMA) in various weight percentages (0%–3%) to improve their antifouling and ultrafiltration properties. The physicochemical properties of the blended membranes, including surface morphology, contact angle, hydrophilicity and surface energy are evaluated. The findings indicate that incorporation PSPMA results in an enhancement of the hydrophilic properties and surface charge of the PES membranes, assessed by employing Bovine Serum Albumin (BSA) as a representative protein. Modified blended membranes display greater Flux Recovery Ratio (FRR%) and exhibit superior fouling resistance. Under the same experimental conditions (0.2 MPa applied pressure), a pure water flux of 154.18 L·m−2·h−1 for PES/PSPMA membrane found substantially greater than pure PES membrane (103.52 L·m−2·h−1) along with Total Fouling Ratio (TFR) of 36% and 64.9% respectively. Exceptional antimicrobial efficacy for modified membranes is revealed against Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus) using disc diffusion technique rendering them well‐suited for water treatment applications. [ABSTRACT FROM AUTHOR]

The underlying mechanisms and biochemical actions responsible for inactivation of pathogenic gram-positive Staphylococcus aureus (S. aureus) and gram-negative Salmonella abony (S. abony) bacteria upon exposure to sub-atmospheric plasma has been investigated. Reduction in colony forming units of the bacteria is established in 60 min and 40 min for S. aureus and S. abony respectively via 6-log reduction curves. The percentage change in reactive oxygen species, such as •OH and H2O2 formed on bacterial membrane during plasma exposure are analysed using spectroflurometer. S. aureus exhibited a significant increase of 324.23% and 1554.84% in •OH and H2O2 radicals respectively. Whereas, 98.14% and 54.49% increase in •OH and H2O2 radicals respectively was observed in S. abony. The oxidation and degradation of DNA is analysed using an ultra violet visible spectrophotometer. The leakage of proteins, lipids, and nucleic acid molecules due to plasma exposure is studied by Attenuated Total Reflectance Fourier-transform infrared spectroscopy (ATR-FTIR). The alteration of secondary protein structure on the cell membrane is observed using Circular Dichroism. Upon exposure to plasma, S. aureus shows a secondary protein structural transition from α-helix (2.4%), β-sheet (78.3%) mixture to modified β-sheet structure (0% α-helix, 79.1% β-sheet). Whereas, S.abony shows a transition from α-helix (1%), β-sheet (64.9%) mixture to modified β-sheet structure (0% α-helix, 74.5% β-sheet). The bacterial morphological study (swelling/shrinking) done using Field Emission Scanning Electron Microscopy (FE-SEM) reveals the deformation of cell membrane. Above findings pave the way for a better understanding of the processes of antimicrobial inactivation strategies when the plasma sterilization process is employed. [ABSTRACT FROM AUTHOR]

Abstract The ability to quickly and accurately analyze Staphylococcus aureus (S. aureus) and isolate the bacteria in a simplified setting is crucial for the early identification and treatment of infectious illnesses. Here, we describe the development of a new aptamer-based detection and separation technique that combines Mg2+-dependent DNAzyme amplification cascades with catalytic hairpin assembly for enhanced sensitivity. This technique uses a rolling circle amplification procedure to build a detection scaffold with a repetitive functional hairpin structure that, upon identifying S. aureus, can launch a catalytic hairpin assembly-mediated DNAzyme-based cascade signal amplification. This allows S. aureus to be isolated using low-speed centrifugation and simultaneously quantified. The approach has a low limit of detection of 21 cfu/mL and a broad detection range of six orders of magnitude due to the inclusion of the catalytic hairpin assembly for signal amplification. In addition to high sensitivity, the method also demonstrates high selectivity for the identification and isolation of S. aureus, making it a useful instrument for reporting S. aureus infections.

IntroductionDuring the COVID-19 Delta variant surge, the CLAIRE cross-sectional study sampled saliva from 120 hospitalized patients, 116 of whom had a positive COVID-19 PCR test. Patients received antibiotics upon admission due to possible secondary bacterial infections, with patients at risk of sepsis receiving broad-spectrum antibiotics (BSA).MethodsThe saliva samples were analyzed with shotgun DNA metagenomics and respiratory RNA virome sequencing. Medical records for the period of hospitalization were obtained for all patients. Once hospitalization outcomes were known, patients were classified based on their COVID-19 disease severity and the antibiotics they received.ResultsOur study reveals that BSA regimens differentially impacted the human salivary microbiome and disease progression. 12 patients died and all of them received BSA. Significant associations were found between the composition of the COVID-19 saliva microbiome and BSA use, between SARS-CoV-2 genome coverage and severity of disease. We also found significant associations between the non-bacterial microbiome and severity of disease, with Candida albicans detected most frequently in critical patients. For patients who did not receive BSA before saliva sampling, our study suggests Staphylococcus aureus as a potential risk factor for sepsis.DiscussionOur results indicate that the course of the infection may be explained by both monitoring antibiotic treatment and profiling a patient’s salivary microbiome, establishing a compelling link between microbiome and the specific antibiotic type and timing of treatment. This approach can aid with emergency room triage and inpatient management but also requires a better understanding of and access to narrow-spectrum agents that target pathogenic bacteria.

The ability to quickly and accurately analyze Staphylococcus aureus (S. aureus) and isolate the bacteria in a simplified setting is crucial for the early identification and treatment of infectious illnesses. Here, we describe the development of a new aptamer-based detection and separation technique that combines Mg2+-dependent DNAzyme amplification cascades with catalytic hairpin assembly for enhanced sensitivity. This technique uses a rolling circle amplification procedure to build a detection scaffold with a repetitive functional hairpin structure that, upon identifying S. aureus, can launch a catalytic hairpin assembly-mediated DNAzyme-based cascade signal amplification. This allows S. aureus to be isolated using low-speed centrifugation and simultaneously quantified. The approach has a low limit of detection of 21 cfu/mL and a broad detection range of six orders of magnitude due to the inclusion of the catalytic hairpin assembly for signal amplification. In addition to high sensitivity, the method also demonstrates high selectivity for the identification and isolation of S. aureus, making it a useful instrument for reporting S. aureus infections. [ABSTRACT FROM AUTHOR]

In this article, we propose a simple photochemical method to synthesize pure La2Ti2O7 films and La2Ti2O7 films doped with silver at 1.0, 3.0, and 5.0 mol%. After annealing the photo-deposited films at 900 °C, XRD, SEM, and XPS analyses showed the formation of a monoclinic La2Ti2O7 phase and the presence of Ag and AgO in doped samples. Photocatalytic tests for Congo red degradation demonstrated that pure La2Ti2O7 achieved 25.4% degradation, while doped samples reached a maximum of 92.7% degradation. Moreover, increasing silver doping on La2Ti2O7 films significantly reduced the growth of Staphylococcus aureus, indicating potential antibacterial properties. The enhanced photoactivity was attributed to the formation of a type I heterojunction between La2Ti2O7 and AgO, and a degradation mechanism was proposed based on Congo red degradation. [ABSTRACT FROM AUTHOR]

ABSTRACT: Avian arthritis is a relatively common disease in the poultry industry, the cause of which is complex. Bacterial arthritis is often caused by infection of Staphylococcus aureus (S. aureus), whereas viral arthritis is caused by avian orthoreovirus (ARV). To investigate the infection of S. aureus and ARV in cases of avian arthritis, a total of 77 samples characterized by arthritis were collected and detection. The results showed that 68.83% of the samples were positive for ARV, and 66.23% were positive for S. aureus. Among them, the ARV mono-infection rate was 22.08%, the S. aureus mono-infection rate was 19.48%, and ARV and S. aureus co-infection rate was 45.45%, indicating that ARV and S. aureus co-infection is common in arthritis cases. To further investigate the synergistic pathogenicity of ARV and S. aureus, ARV and S. aureus were used to mono-infect, co-infect, and (or) sequential infect SPF chickens and the clinical indications, pathologic changes, ARV load, S. aureus bacterial distribution, and cytokine level of the challenged chickens were evaluated. Decreased weight gain, increased mortality, and difficulties in standing were observed in all dual-infected groups and the singular-infected group. There were significantly more severe macroscopic and microscopic hock lesions, and larger amounts of a wider range of tissue distribution of ARV antigens and S. aureus bacterial distribution in the dual-infected groups compared to the single-infected and control groups. Cytokine detection showed a significant change in IFN-γ, IL-1β, and IL-6 levels in the infected groups, especially in the ARV–S. aureus co-infection, and (or) sequential infection groups, compared with the control group. Hence, ARV and S. aureus synergistically increased mortality in infected chickens, potentiated the severity of arthritis, and increased the amount of ARV RNA in tendons.

In the present work, zinc sulphide (ZnS) nanoparticles are synthesized via solvothermal route using different capping agents. The sphalerite phase is detected by X-ray diffraction (XRD) analysis. The morphology of the nanoparticles is confirmed by scanning electron microscopy (SEM) and high resolution transmission electron microscopy (HRTEM). The average particle size is found between 5 and 7 nm as calculated by HRTEM analysis. Energy dispersive X-ray spectroscopy (EDX) specified the desired elemental composition. Bandgap studies are done by UV–visible absorption and the photoluminescence spectroscopy (PL) gives the emission peaks centered at 452 nm and 460 nm. Fourier transform infra-red spectroscopy (FTIR) of ZnS nanoparticles confirmed the presence of various functional groups and showed the encapsulation of capping agents. The cylindrical non-woven network of PAN-ZnS nanofibers synthesized by electrospinning technique as confirmed by SEM and the distribution of nanoparticles in the polymer matrix is confirmed by HRTEM analysis. EDX and FTIR showed the incorporation of ZnS nanoparticles in polymer matrix. The objective of the present work is to synthesize the PAN-ZnS composite nanofibers for antibacterial applications. To the best of our knowledge this type of studies using polyacrylonitrile (PAN) as a polymer are not found in the literature showing the novelty of our work. [ABSTRACT FROM AUTHOR]

For decades now, DNA fingerprinting by means of pulsed-field gel electrophoresis (PFGE) continues to be the most widely used to separate large DNA molecules and distinguish between different strains in alternating pulses. This is done by isolating intact chromosomal DNA and using restriction enzymes with specific restriction sites to generate less than 30 restriction fragments from 50 Kb to 10 Mbp. These results make clone-specific band profiles easy to compare. Specialized equipment is required for the optimization of DNA separation and resolution, among which a contour-clamped homogeneous electric field (CHEF) apparatus is the most commonly used. As a result, the PFGE analysis of a bacterial genome provides useful information in terms of epidemiological investigations of different bacterial pathogens. For Staphylococcus aureus subtyping, despite its limitations and the emergence of alternative methods, PFGE analysis has proven to be an adequate choice and the gold standard for determining genetic relatedness, especially in outbreak detection and short-term surveillance in the veterinary field. [ABSTRACT FROM AUTHOR]

Antimicrobial resistance is a global health threat and efforts to mitigate it is warranted, thus the need for local antibiograms to improve stewardship. This study highlights the process that was used to develop an antibiogram to monitor resistance at a secondary-level health facility to aid empirical clinical decision making in a sub-Saharan African county. This retrospective cross-sectional descriptive study used 3 years of cumulative data from January 2016 to December 2018. Phenotypic data was manually imputed into WHONET and the cumulative antibiogram constructed using standardized methodologies according to CLSI M39-A4 guidelines. Pathogens were identified by standard manual microbiological methods and antimicrobial susceptibility testing was performed using Kirby-Bauer disc diffusion method according to CLSI M100 guidelines. A total of 14,776 non-duplicate samples were processed of which 1163 (7.9%) were positive for clinically significant pathogens. Among the 1163 pathogens, E. coli (n = 315) S. aureus (n = 232), and K. pneumoniae (n = 96) were the leading cause of disease. Overall, the susceptibility for E. coli and K. pneumoniae from all samples were: trimethoprim-sulfamethoxazole (17% and 28%), tetracycline (26% and 33%), gentamicin (72% and 46%), chloramphenicol (76 and 60%), and ciprofloxacin (69% and 59%), and amoxicillin/clavulanic (77% and 54%) respectively. Extended spectrum beta-lactamase (ESBL) resistance was present in 23% (71/315) vs. 35% (34/96) respectively. S. aureus susceptibility for methicillin was 99%. This antibiogram has shown that improvement in combination therapy is warranted in The Gambia. [ABSTRACT FROM AUTHOR]

Newborns are highly susceptible to infections; however, the underlying mechanisms that regulate the anti-microbial T-helper cells shortly after birth remain incompletely understood. To address neonatal antigen-specific human T-cell responses against bacteria, Staphylococcus aureus (S. aureus) was used as a model pathogen and comparatively analyzed in terms of the polyclonal staphylococcal enterotoxin B (SEB) superantigen responses. Here, we report that neonatal CD4 T-cells perform activation-induced events upon S. aureus/APC-encounter including the expression of CD40L and PD-1, as well as the production of Th1 cytokines, concomitant to T-cell proliferation. The application of a multiple regression analysis revealed that the proliferation of neonatal T-helper cells was determined by sex, IL-2 receptor expression and the impact of the PD-1/PD-L1 blockade. Indeed, the treatment of S. aureus-activated neonatal T-helper cells with PD-1 and PD-L1 blocking antibodies revealed the specific regulation of the immediate neonatal T-cell responses with respect to the proliferation and frequencies of IFNγ producers, which resembled in part the response of adults' memory T-cells. Intriguingly, the generation of multifunctional T-helper cells was regulated by the PD-1/PD-L1 axis exclusively in the neonatal CD4 T-cell lineage. Together, albeit missing memory T-cells in neonates, their unexperienced CD4 T-cells are well adapted to mount immediate and strong anti-bacterial responses that are tightly controlled by the PD-1/PD-L1 axis, thereby resembling the regulation of recalled memory T-cells of adults. [ABSTRACT FROM AUTHOR]

Infective endocarditis (IE) is a severe illness characterized by vegetation of bacterial thrombosis. We hypothesized that adding recombinant tissue-type plasminogen activator (rt-PA) to antibiotics would contribute to good results in the treatment of IE. As an in vitro study, we injected labeled Staphylococcus aureus (S. aureus) and either rt-PA or PBS + plasminogen into a polydimethylsiloxane flow chamber with fibrin on a coverslip, and then performed immunofluorescent area assessment. As an in vivo experiment, IE model rats that had suffered mechanical damage in the aortic valve by catheter and revealed bacterial vegetation caused by S. aureus injection were treated with either a control, cefazolin (CEZ), rt-PA, or rt-PA + CEZ, for 7 days. Survival was assessed for 14 days after the appearance of vegetation, with daily monitoring of the vegetation by transthoracic echocardiography (TTE). The in vitro investigation showed that perfusion of rt-PA could detach S. aureus significantly more efficiently than PBS could. In the in vivo research, the rt-PA + CEZ group survived significantly longer than the other groups, and rt-PA + CEZ was more effective than CEZ in the dissolution of vegetation, as observed by TTE. In conclusion, adding rt-PA to antibiotic treatment could dissolve the vegetation component synergistically and improve the survival rate. [Display omitted] • Our in vitro study showed that rt-PA detaches the bacteria-fibrin connection. • Rt-PA + cefazolin improved the survival rate in staphylococcal IE model rats. • Rt-PA + cefazolin was effective in dissolving and sterilizing the vegetation. [ABSTRACT FROM AUTHOR]

Sonodynamic antimicrobial chemotherapy (SACT) is an effective antimicrobial treatment that can avoid the production of drug-resistant bacteria. Design and development of new high-efficiency sonosensitizers play a key role in the practical application of SACT. The bacteriostatic effects of two phenothiazine compounds, toluidine blue (TB) and azure A (AA) combined with ultrasonic (US) on Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus) were studied, and the sonodynamic antibacterial activities of TB and AA were compared. The reactive oxygen species (ROS) and the types of ROS produced in the sonodynamic system were detected and the sonodynamic mechanisms of TB and AA were proposed. The sonodynamic bacteriostasis mediated by TB and AA increased with the increasing concentration of sonosensitizer, the extension of sonication time and the increase of reaction temperature. The production of ROS was the main reason that TB and AA had excellent sonodynamic antibacterial performance. Singlet oxygen (1O 2) and hydroxyl radical (•OH) were the main ROS types in the sonodynamic antibacterial system. The ROS produced by the combined action of AA and US was higher than that of TB. Both TB and AA displayed excellent sonodynamic antibacterial activities. Moreover, AA had a higher sonodynamic activity than TB. The electron donation effect and steric hindrance effect of the methyl group of phenothiazine parent nucleus of TB might be the cause of the decrease of its sonodynamic activity. These results would provide a valuable reference for the further study of phenothiazines sonosensitizers and their clinical application in SACT. • TB and AA had excellent sonodynamic antibacterial performance. • 1O 2 and •OH were the main ROS in the sonodynamic antibacterial process. • The sonodynamic activity of AA was higher than that of TB. [ABSTRACT FROM AUTHOR]

Vat photopolymerization (VPP) is an additive manufacturing process commonly used in medical applications. This work aims, for the first time in the literature, to extend and enhance the performance of a commercial medical-grade resin for the VPP process, with the development of nanocomposites, using Copper (Cu) nanoparticles as the additive at two different concentrations. The addition of the Cu nanoparticles was expected to enhance the mechanical properties of the resin and to enable biocidal properties on the nanocomposites since Cu is known for its antibacterial performance. The effect of the Cu concentration was investigated. The nanocomposites were prepared with high-shear stirring. Specimens were 3D printed following international standards for mechanical testing. Their thermal and spectroscopic response was also investigated. The morphological characteristics were examined. The antibacterial performance was evaluated with an agar well diffusion screening process. The experimental results were analyzed with statistical modeling tools with two control parameters (three levels each) and eleven response parameters. Cu enhanced the mechanical properties in all cases studied. 0.5 wt.% Cu nanocomposite showed the highest improvement (approximately 11% in tensile and 10% in flexural strength). The antibacterial performance was sufficient against S. aureus and marginal against E. coli. [ABSTRACT FROM AUTHOR]

Oleate hydratase (OhyA), a flavoenzyme that catalyzes the hydration of unsaturated fatty acids, has been identified in various Bacillales organisms, including those in the Listeria , Lysinibacillus , Paenibacillus , and Staphylococcus genera. In this study, we combine structural biology with molecular and phylogenetic analyses to investigate the evolutionary dynamics of the OhyA protein family within the Bacillales order. Our evolutionary analysis reveals two distinct OhyA clades (clade I and clade II) within Bacillales that, while sharing catalytic function, exhibit significant genomic and structural differences. Our findings suggest that these OhyA clades originated from independent evolutionary processes through convergent evolution rather than gene duplication. We also show that the evolutionary divergence in OhyA is likely due to intrinsic sequence variations rather than being strictly linked to functional domain changes. Furthermore, within the Staphylococcus genus, we observed that the evolution of the ohyA gene aligns with the species tree, supporting a common ancestral origin. This study enhances our understanding of the impact of evolutionary history on the structure and function of OhyA across the Bacillales order., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Neff, Lages, Donworth, Brien and Radka.)

Staphylococcus aureus (S. aureus), especially methicillin-resistant S. aureus (MRSA), causes wound infections, whose treatment remains a clinical challenge. Bacterium-infected wounds often create acidic niches with a pH 4.5-6.5. Endolysin LysSYL, which is derived from phage SYL, shows promise as an antistaphylococcal agent. However, endolysins generally exhibit instability and possess low bioavailability in acidic microenvironments. Here, an array of self-assembling peptides is designed, and peptide L5 is screened out based on its gel formation property and bioavailability. L5 exerted a pH-switchable antimicrobial effect (pH 5.5) and formed biocompatible hydrogels at neutral pH (pH 7.4). The LysSYL-loaded L5 can assemble L5@LysSYL hydrogels, increase thermal stability, and exhibit the slow-release effect of LysSYL. Effective elimination of S. aureus is achieved by L5@LysSYL through bacterial membrane disruption and cell separation inhibition. Moreover, L5@LysSYL hydrogels exhibit great potential in promoting wound healing in a mouse wound model infected by MRSA. Furthermore, L5@LysSYL hydrogels are safe and can decrease the cytokine levels and increase the number of key factors for vessel formation, which contribute to wound healing. Overall, the self-assembling L5@LysSYL can effectively clean MRSA and promote wound healing, which suggests its potential as a pH-sensitive wound dressing for the management of wound infections., (© 2024 The Author(s). Advanced Materials published by Wiley‐VCH GmbH.)

Background: To the present day infective endocarditis (IE) represents a life-threatening disease with high mortality rate especially when caused by Staphylococcus aureus (S. aureus), the most common causative pathogen in this disease. Diagnosis of IE is based on clinical manifestations, pathogen detection by blood cultures and echocardiographic or other imaging findings. However, none of the methods used is capable of detecting the causative bacterial cells on the endothelium directly. Modern molecular imaging such as positron emission tomography/computed tomography (PET/CT) is playing an increasingly important role in unclear IE cases. This study focused on 2-[18F]F-p-aminobenzoic acid (2-[18F]F-PABA), a bacteria specific tracer for the diagnosis of IE using PET imaging for direct pathogen detection., Methods: In vitro assays were performed to analyze 2-[18F]F-PABA uptake by S. aureus. For proof-of-concept in vivo trials an endocarditis mouse model was used to diagnose IE by PET/Magnetic resonance (MR) imaging. A subcutaneous abscess mouse model was supplemented to create larger bacterial vegetations for PET imaging., Results: 2-[18F]F-PABA in vitro uptake by S. aureus was confirmed. Only living bacteria were able to accumulate the tracer while the extent of uptake varied between different S. aureus strains. In the in vivo proof-of-concept, IE was visualized in mice using 2-[18F]F-PABA-PET/MR imaging. Subsequently, 2-[18F]F-PABA specifically located S. aureus vegetations in the subcutaneous abscess model., Conclusions: This study highlights the great potential of 2-[18F]F-PABA imaging for the direct detection of IE. Future studies might further investigate the clinical potential of this molecular imaging approach, finally aiming at a clinical implementation., (© The Author(s) 2024. Published by Oxford University Press on behalf of Infectious Diseases Society of America.)

Hyper-IgE syndromes (HIES) are a group of inborn errors of immunity (IEI) caused by monogenic defects such as in the gene STAT3 (STAT3-HIES). Patients suffering from HIES show an increased susceptibility to Staphylococcus aureus (S. aureus) including skin abscesses and pulmonary infections. To assess if the underlying immune defect of STAT3-HIES patients influences the resistance patterns, pathogenicity factors or strain types of S. aureus. We characterized eleven S. aureus strains isolated from STAT3-HIES patients (n = 4) by whole genome sequencing (WGS) to determine presence of resistance and virulence genes. Additionally, we used multi-locus sequence typing (MLST) and protein A (spa) typing to classify these isolates. Bacterial isolates collected from this cohort of STAT3-HIES patients were identified as common spa types in Germany. Only one of the isolates was classified as methicillin-resistant S. aureus (MRSA). For one STAT3 patient WGS illustrated that infection and colonization occurred with different S. aureus isolates rather than one particular clone. The identified S. aureus carriage profile on a molecular level suggests that S. aureus strain type in STAT3-HIES patients is determined by local epidemiology rather than the underlying immune defect highlighting the importance of microbiological assessment prior to antibiotic treatment. [ABSTRACT FROM AUTHOR]

The rapid spread of staphylococcus aureus (S. aureus) causes an increased morbidity and mortality, as well as great economic losses in the world. Anti-S. aureus infection becomes a major challenge for clinicians and nursing professionals to address drug resistance. Hence, it is urgent to explore high efficiency, low toxicity, and environmental-friendly methods against S. aureus. Metal-organic frameworks (MOFs) represent great potential in treating S. aureus infection due to the unique features of MOFs including tunable chemical constitute, open crystalline structure, and high specific surface area. Especially, these properties endow MOF-based materials outstanding antibacterial effect, which can be mainly attributed to the continuously released active components and the exerted catalytic activity to fight bacterial infection. Herein, the structural characteristics of MOFs and evaluation method of antimicrobial activity are briefly summarized. Then we systematically give an overview on their recent progress on antibacterial mechanisms, metal ion sustained-release system, controlled delivery system, catalytic system, and energy conversion system based on MOF materials. Finally, suggestions and direction for future research to develop and mechanism understand MOF-based materials are discussed in antibacterial application. [ABSTRACT FROM AUTHOR]

Staphylococcus aureus (S. aureus) presents a significant challenge in both nosocomial and community settings due to its pathogenicity. The emergence of drug-resistant strains exacerbates S. aureus infections, leading to increased mortality rates. PyrG, a member of the cytidine triphosphate (CTP) synthase family, serves as a crucial therapeutic target against S. aureus due to the pivotal role of CTP in cellular metabolism. However, the structural and mechanistic details of S. aureus PyrG remains unknown. Here, we successfully expressed and purified monomeric PyrG. Mutational experiments were conducted based on the results of molecular docking. Based on the results of the molecular docking, we carried out mutation experiments and found that Q386A dramatically decreased the CTP synthase activity compared to the wild-type protein, while Y54A almost completely abolished the activity. Exposure of S. aureus to the kinase inhibitor crizotinib increased expression of gene pyrG. Our results identify the two key sites on PyrG for the CTP synthase activity, and present PyrG gene expression increased during the treatment of crizotinib, which may eventually provide valuable guidance for the development of new drugs against S. aureus infections. • Monomeric PyrG was obtained. • Mutagenesis of residues Q386 or Y54 to alanine decreased the activity of PyrG compared to wild-type protein. • The expression of the pyrG gene increased when S. aureus was exposed to crizotinib. [ABSTRACT FROM AUTHOR]

Multi-resistant Staphylococcus aureus (S. aureus) infection is a significant global health concern owing to its high mortality and morbidity rates. Coagulase (Coa), a key enzyme that activates prothrombin to initiate host coagulation, has emerged as a promising target for anti-infective therapeutic approaches. This study identified sinigrin as a potent Coa inhibitor that significantly inhibited S. aureus -induced coagulation at concentration as low as 32 mg/L. Additionally, at a higher concentration of 128 mg/L, sinigrin disrupted the self-protection mechanism of S. aureus. Thermal shift and fluorescence-quenching assays confirmed the direct binding of sinigrin to the Coa protein. Molecular docking analysis predicted specific binding sites for sinigrin in the Coa molecule, and point mutation experiments highlighted the importance of Arg-187 and Asp-222 as critical binding sites for both Coa and sinigrin. In vivo studies demonstrated that the combination of sinigrin with oxacillin exhibited greater antibacterial efficacy than oxacillin alone in the treatment of S. aureus -induced pneumonia in mice. Furthermore, sinigrin was shown to reduce bacterial counts and inflammatory cytokine levels in the lung tissues of S. aureus -infected mice. In summary, sinigrin was shown to directly target Coa, resulting in the attenuation of S. aureus virulence, which suggests the potential of sinigrin as an adjuvant for future antimicrobial therapies. [ABSTRACT FROM AUTHOR]

Staphylococcus aureus (S. aureus) is a prevalent bacterium that affects patient recovery post-surgery and can cause postoperative complications. Consequently, detecting S. aureus in clinical settings is critical for ensuring patient well-being. In our work, a "visual-colorimetric-photothermal" three-in-one multimodal lateral flow immunoassays (LFIA) was developed for S. aureus detection to meet the growing demand for point-of-care testing (POCT) of bacteria in the field. The sensors relied on Pd/Pt@Ab1 NPs nanoprobes, created by decorating the surface of bimetallic nanoprobes (Pd/Pt NPs) with S. aureus antibodies (Ab1). A visual detection mode was realized by the Pd/Pt@Ab1 NPs being captured on the surface of test line (T line) using the sandwich method, offering a limit of detection (LOD) of 103 CFU/mL. Subsequently, Pd/Pt@Ab1 NPs catalyzed the TMB system into oxTMB to achieve colorimetric analysis, covering a detection range from 102 to 107 CFU/mL. Remarkably, the oxTMB exhibited the photothermal conversion efficiency under near-infrared (NIR) light, leading to temperature variations under 808 nm irradiation, correlating with different bacteria concentrations, with an LOD as low as 4 CFU/mL. The methodology enabled quantitative analysis of bacteria in clinical samples with satisfactory results. The diversity, complementarity, and synergy of the integrated output signals in this multimode LFIA improved the flexibility, utility, and accuracy of the assay, performing well as a POCT platform for various application scenarios. • A "three-in-one" multimode lateral flow immunoassay was developed for the detection of Staphylococcus aureus in clinic. • Pd/Pt@Ab1 NPs was synthesized to enhance and improve the sensitivity of bacterial detection. • Oxidized TMB had colorimetric and photothermal properties. • The methodology developed exhibited flexibility, practicality and accuracy. [ABSTRACT FROM AUTHOR]

Antimicrobial resistance is a global health threat and efforts to mitigate it is warranted, thus the need for local antibiograms to improve stewardship. This study highlights the process that was used to develop an antibiogram to monitor resistance at a secondary-level health facility to aid empirical clinical decision making in a sub-Saharan African county. This retrospective cross-sectional descriptive study used 3 years of cumulative data from January 2016 to December 2018. Phenotypic data was manually imputed into WHONET and the cumulative antibiogram constructed using standardized methodologies according to CLSI M39-A4 guidelines. Pathogens were identified by standard manual microbiological methods and antimicrobial susceptibility testing was performed using Kirby-Bauer disc diffusion method according to CLSI M100 guidelines. A total of 14,776 non-duplicate samples were processed of which 1163 (7.9%) were positive for clinically significant pathogens. Among the 1163 pathogens, E. coli (n = 315) S. aureus (n = 232), and K. pneumoniae (n = 96) were the leading cause of disease. Overall, the susceptibility for E. coli and K. pneumoniae from all samples were: trimethoprim-sulfamethoxazole (17% and 28%), tetracycline (26% and 33%), gentamicin (72% and 46%), chloramphenicol (76 and 60%), and ciprofloxacin (69% and 59%), and amoxicillin/clavulanic (77% and 54%) respectively. Extended spectrum beta-lactamase (ESBL) resistance was present in 23% (71/315) vs. 35% (34/96) respectively. S. aureus susceptibility for methicillin was 99%. This antibiogram has shown that improvement in combination therapy is warranted in The Gambia.

Staphylococcus aureus (S. aureus) infection is difficult to fight, previous experimental reports have demonstrated thioridazine (TZ) and tetracycline (TC) is an inhibitor of S. aureus efflux pump NorA and autolysin Atl, respectively, here, by means of molecular docking and molecular dynamics simulation, we observed that thioridazine (TZ) and tetracycline (TC) blocked the binding of substrates to NorA and Atl, respectively, and reduced their activities, and our antibacterial susceptibility test and three-dimensional checkerboard method showed that the three-drug combination of antibiotic cloxacillin (CXN), TZ and TC had a synergistic anti-Staphylococcal activity in vitro , and α-Hemolysin tests and scanning electron microscopy showed that the three-drug combination and the subinhibitory concentration of the combination significantly inhibited the secretion of α-hemolysin relative to the number of membrane-derived vesicles produced by S. aureus. Whereas Western blot and pharmacological inhibition assays showed that the three-drug combination significantly inhibited the expression of MAPK/NF-κB/NLRP3 proteins in macrophages induced with S. aureus α-hemolysin. In vivo , the drug combination significantly reduced bacterial colony-forming unit counts in the viscera of a mouse peritonitis model of S. aureus infection, therapy reduced the primary inflammatory pathology and the bacteria-stimulated release of cytokines such as IL-1β and TNF-α, and inhibited the expression of MAPK/NF-κB/NLRP3 proteins in peritoneal macrophages. Thus, the combination of efflux pump inhibitor, autolysis inhibitor and antibiotic, is a novel anti-Staphylococcal and anti-inflammatory strategy who owning good antibacterial activity and significant inhibiting staphylococcal α-hemolysin and inflammation. [ABSTRACT FROM AUTHOR]

Donya Jamshidi, Mohammad Reza Sazegar Department of Chemistry, Islamic Azad University, Tehran, IranCorrespondence: Mohammad Reza SazegarDepartment of Chemistry, Islamic Azad University, North Tehran Branch, Hakimiyeh, Tehran, IranTel +98-9381199151Email m_r_sazegar@yahoo.comIntroduction: A novel biocomposite chitosan/graphite based on zinc-grafted mesoporous silica nanoparticles (CGZM-bio) was synthesized and the antibacterial activities of this compound along with that of Zn-MSN nanoparticles were investigated.Methods: The CGZM-bio biocomposite was synthesized using sol–gel and post-synthesis method under UV radiation. The characterizations of the samples were carried out using FTIR, XRD, SEM, and nitrogen adsorption and desorption. The antibacterial activity was carried out against Staphylococcus aureus (S. aureus) and Escherichia coli (E. coli) after 18 h at 310 K.Results: The suspension samples of the Zn-MSN and CGZM-bio (2– 100 μg.mL− 1) presented antibacterial activities against S. aureus and E. coli. The minimum inhibitory concentration (MIC) values against E. coli for the Zn-MSN and CGZM-bio samples were 10 and 5 μg.mL− 1, respectively, while the MIC against S. aureus for both nanomaterials was 10 μg.mL− 1.Discussion: The antibacterial activities of these materials are due to the generation of radical oxygen species such as •OH, H2O2, and O2 2- during the UV radiation via the generation of the electron–hole pairs which in turn damage the bacteria cells. These nanomaterials may be used in biomedical devices as antibacterial agents.Keywords: Zn-MSN, Staphylococcus aureus, Escherichia coli, antimicrobial activity, UV radiation, electron–hole pair

Vat photopolymerization (VPP) is an additive manufacturing process commonly used in medical applications. This work aims, for the first time in the literature, to extend and enhance the performance of a commercial medical-grade resin for the VPP process, with the development of nanocomposites, using Copper (Cu) nanoparticles as the additive at two different concentrations. The addition of the Cu nanoparticles was expected to enhance the mechanical properties of the resin and to enable biocidal properties on the nanocomposites since Cu is known for its antibacterial performance. The effect of the Cu concentration was investigated. The nanocomposites were prepared with high-shear stirring. Specimens were 3D printed following international standards for mechanical testing. Their thermal and spectroscopic response was also investigated. The morphological characteristics were examined. The antibacterial performance was evaluated with an agar well diffusion screening process. The experimental results were analyzed with statistical modeling tools with two control parameters (three levels each) and eleven response parameters. Cu enhanced the mechanical properties in all cases studied. 0.5 wt.% Cu nanocomposite showed the highest improvement (approximately 11% in tensile and 10% in flexural strength). The antibacterial performance was sufficient against S. aureus and marginal against E. coli.

Analysis of mecA gene in Staphylococcus aureus (S. aureus) is essential for controlling infections in intensive care units (ICU) and preventing the use of ineffectual empirical treatments. However, quantitative determination of the mecA gene remains difficult. Herein, we propose a simple and sensitive colorimetric approach by integrating exonuclease-III (Exo-III) assisted signal cascade and G-quadruplex/hemin DNAzymes (G4 DNAzymes) catalyzed 2,2′-azino-bis (3-ethylben-zothiazoline-6-sulfonic acid) (ABTS) based color reaction. In this method, signal amplification does not necessitate the use of complex experimental components, such as multiple enzymes and primer design, while still maintaining a high signal amplifying efficiency. Therefore, the method has a broad mecA gene detection range from 10 fM to 1 nM and a low limit of detection down to 3.4 fM level. Taking the merit of simplicity and high sensitivity, the approach is promising in analyzing mecA gene in S. aureus and diagnosing infections. [Display omitted] • Analysis of mecA gene in Staphylococcus aureus (S. aureus) is essential for controlling infections. • We propose a simple and sensitive colorimetric approach by integrating exonuclease-III assisted signal cascade and G-quadruplex/hemin based color reaction. • The method does not require complicated experimental components (e.g. enzymes and primer design) and preparation for signal amplification. [ABSTRACT FROM AUTHOR]

Abstract Background Methicillin-resistant Staphylococcus aureus (MRSA) causes serious infections in hospitals. Penthorum chinense Pursh (PCP), employed by the Miao ethnic minority in China, presents antibacterial activities. In this study, the anti-Staphylococcus aureus activities in the pinocembrin-7-O residue-rich fraction from PCP (PGF) were evaluated and characterized. Methods The PGF was prepared with 70% ethanol reflux extraction followed by fractional extraction and column chromatography. Pinocembrin-7-O residue components were identified with electrospray ionization mass spectrometry (ESI-MS). Anti-S. aureus activities of the fraction and the main components were evaluated in vitro with serially diluted microbroth assays. Cytotoxicity was evaluated with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) chromogenic assays using the NCTC 1469 cell line. Results This study indicated that the PGF and three components (S1, S2, and S3) presented anti-S. aureus activities, including against clinically isolated MRSA strains. The molecular masses of S1, S2, and S3 were identical to those of pinocembrin-7-O-[4″,6″-hexahydroxydiphenoyl (HHDP)]-β-D-glucose, pinocembrin-7-O-[3″-O-galloyl-4″,6″-(s)-HHDP]-β-D-glucose, and Thonningianin A, respectively. The PGF, S1, S2, and S3 all presented an identical minimum inhibitory concentration (MIC) against S. aureus ATCC 25923 and ATCC 43300, which was 62.5 μg/mL. The minimum bactericidal concentrations (MBCs) of the PGF and S3 against ATCC 25923 were 125 and 250 μg/mL, and the MBCs of the PGF, S2, and S3 against ATCC 43300 were 250, 500, and 250 μg/mL, respectively. A time-kill assay consistently indicated that none of the bacterial clones of ATCC 25923 and ATCC 43300 could survive under 2× and 4× MIC PGF treatment for 24 h, respectively. In contrast, 104 CFU (colony-forming units) of ATCC 25923 and ATCC 43300 were killed by 8× and 4× MIC S3 within 24 h, respectively. Additionally, 1×, 2×, and 4× MIC the PGF presented similar postantibiotic effects (PAEs) on the strain ATCC 25923. However, the PAE of the PGF on the strain ATCC 43300 was concentration dependent (1×

Staphylococcus aureus (S. aureus) is a pathogenic gram-positive bacterium that normally resides in the skin and nose of the human body. It is subject to fluctuations in environmental conditions that may affect the integrity of the membrane. S. aureus produces carotenoids, which act as antioxidants. However, these carotenoids have also been implicated in modulating the biophysical properties of the membrane. Here, we investigate how carotenoids modulate the thermotropic phase behavior of model systems that mimic the phospholipid composition of S. aureus. We found that carotenoids depress the main phase transition of DMPG and CL, indicating that they strongly affect cooperativity of membrane lipids in their gel phase. In addition, carotenoids modulate the phase behavior of mixtures of DMPG and CL, indicating that they may play a role in modulation of lipid domain formation in S. aureus membranes.

Globally, prematurity is the leading cause of neonatal mortality (babies in the first four weeks of life) and now the second leading cause of mortality after pneumonia in children under age five. The neonatal gut microbial colonization is crucial in the human life cycle. Placental microbiota transmits from the gut microbiota plays a significant role in association with kinship. Simultaneously, this transition is being made from mother to infant. This comparative study explored the diversity of microbiota associated with term and preterm neonates by evaluating the placental samples. The study found that 16/68 (23.5%) full-term placental samples were positive for S. aureus; on the other hand, 4/16 (25%) preterm placental samples confirmed culture growth for S. aureus. Antimicrobial susceptibility patterns showed that Staphylococcusaureus (S. aureus) isolates from both types of samples were resistant to Ofloxacin, Trimethoprim-sulfamethoxazole, Oxacillin, and Cefoxitin. However, Methicillin-Resistant Staphylococcus aureus (MRSA) detection was 43.75% in full-term and 75% in preterm placental samples. Moreover, two isolates were positive for both mecA and PVL virulent genes, and the rest were positive only for the mecA gene. Interestingly few isolates lacked both characteristic MRSA genes, mecA and PVL. Notably, resistances were more inclined towards preterm samples for antimicrobial susceptibility and MRSA screening. It may be concluded that there is a significant presence of S. aureus in the placenta of mothers with term and preterm deliveries which might be responsible for preterm deliveries. Therefore, judicious use of antibiotics during pregnancies may help prevent preterm births.

The growing numbers of pathogenic bacteria acquiring multidrug resistance are posing the challenges to develop novel therapeutic approaches. Photodynamic bacterial inactivation assaults multiple subcellular targets of pathogenic bacteria, including Staphylococcus aureus, with low tendency to induce further resistance. A wide range of aPDI photosensitizers were investigated for localized infections, including endogenous porphyrins and encapsulated sensitizers in nanoparticles. This mini review presents the concepts of aPDI, a field consisting of more than 3000 research articles published over 30 years. Mutated PBP membrane enzymes of multiantibiotic resistant S. aureus are plausible targets of aPDI. Synergistic aPDI using simultaneously DP, light irradiation and the antibiotic oxacillin, was recently described, eliminating temporarily oxacillin resistance. Additive antimicrobial effects were achieved with DP‐aPDI combined with additional classes of antibiotics. We propose that mutual aPDI adjuvant with antibiotics or other toxic molecules is opening a new therapeutic window for topical life‐threatening infections, without induction of further resistance. [ABSTRACT FROM AUTHOR]

Staphylococcus aureus (S. aureus), a Gram-positive bacterium, causes a wide range of infections, and diagnosis at an early stage is challenging. Targeting the maltodextrin transporter has emerged as a promising strategy for imaging bacteria and has been able to image a wide range of bacteria including S. aureus. However, little is known about the maltodextrin transporter in S. aureus , and this prevents new S. aureus specific ligands for the maltodextrin transporter from being developed. In Gram-positive bacteria, including S. aureus , the first step of maltodextrin transport is the binding of the maltodextrin-binding protein malE to maltodextrins. Thus, understanding the binding affinity and characteristics of malE from S. aureus is important to developing efficient maltodextrin-based imaging probes. We evaluated the affinity of malE of S. aureus to maltodextrins of various lengths. MalE of S. aureus (SAmalE) was expressed in E. coli BL21(DE3) and purified by Ni-NTA resin. The affinities of SAmalE to maltodextrins were evaluated with isothermal titration calorimetry. SAmalE has low affinity to maltose but binds to maltotriose and longer maltodextrins up to maltoheptaose with affinities up to Ka = 9.02 ± 0.49 × 105 M−1. SAmalE binding to maltotriose-maltoheptaose was exothermic and fit a single-binding site model. The van't Hoff enthalpy in the binding reaction of SAmalE with maltotriose was 9.9 ± 1.3 kcal/mol, and the highest affinity of SAmalE was observed with maltotetraose with Ka = 9.02 ± 0.49 × 105 M−1. In the plot of ΔH-T*ΔS, the of Enthalpy−Entropy Compensation effect was observed in binding reaction of SAmalE to maltodextrins. Acarbose and maltotetraiol bind with SAmalE indicating that SAmalE is tolerant of modifications on both the reducing and non-reducing ends of maltodextrins. Our results show that unlike ECmalE and similar to the maltodextrin binding protein of Streptococci , SAmalE primarily binds to maltodextrins via hydrogen bonds. This is distinct from the maltodextrin binding protein of Streptococci , SAmalE that binds to maltotetraiol with high affinity. Understanding the binding characteristics and tolerance to maltodextrins modifications by maltodextrin binding proteins will hopefully provide the basis for developing bacterial species-specific maltodextrin-based imaging probes. • MalE of S. aures (SAmalE) indicates highest affinity to maltotetraose. • SAmalE binds maltodextrins longer than maltotriose. • The binding reaction with maltodextrins is monophasic exothermic reaction. • SAmalE is tolerant for modifications in both the reducing and non-reducing ends. [ABSTRACT FROM AUTHOR]

Staphylococcus aureus is a causative agent of bovine mastitis, capable of causing significant economic losses to the dairy industry worldwide. This study focuses on obtaining single-chain fragment variables (scFvs) against the virulence factors of S. aureus and evaluates the protective effect of scFvs on bovine mammary epithelial (MAC-T) cells and mice mammary gland tissues infected by S. aureus. After five rounds of bio-panning, four scFvs targeting four virulence factors of S. aureus were obtained. The complementarity-determining regions (CDRs) of these scFvs exhibited significant diversities, especially CDR3 of the VH domain. In vitro, each of scFvs was capable of inhibiting S. aureus growth and reducing the damage of MAC-T cells infected by S. aureus. Preincubation of MAC-T cells with scFvs could significantly attenuate the effect of apoptosis and necrosis compared with the negative control group. In vivo, the qPCR and ELISA results demonstrated that scFvs reduced the transcription and expression of Tumor Necrosis Factor alpha (TNF-α), interleukin-1β (IL-1β), IL-6, IL-8, and IL-18. Histopathology and myeloperoxidase (MPO) results showed that scFvs ameliorated the histopathological damages and reduced the inflammatory cells infiltration. The overall results demonstrated the positive anti-inflammatory effect of scFvs, revealing the potential role of scFvs in the prevention and treatment of S. aureus infections.

Abstract Staphylococcus aureus (S. aureus) is one of the most common pathogens causing both human and animal infections. Transmission of S. aureus to humans via contaminated food continues to be a health public concern. In the present study, 104 strains including eight MRSA strains were identified from 507 chicken samples (20.5%) in three cities of China. These strains harbored the highest resistance against penicillin (91.4%), followed by tetracycline (64.4%), erythromycin (53.5%), and kanamycin (32.7%). We used spa typing to classify these isolates into 28 types belonging to two lineages including the predominant type t112 (n = 26) and four newly identified spa types. Among the 104 strains, six carried the CRISPR-Cas system, a prokaryotic immune system which protects against foreign genetic elements. Interestingly, rpoB gene sequencing demonstrated that these six initially designated ST2250 strains were in fact S. argenteus , a novel Staphylococcus species genetically closely related to S. aureus. Three Staphylococcus CRIPSR types containing ten spacers identified in these strains have been reported in CRISPR-positive S. aureus. Additionally, 80% of the spacers showed homology to S. aureus phages demonstrating that these conserved spacers were closely related to the phages in the environment of S. argenteus. We speculated that the identical CRISPR types and spacers in both S. argenteus and S. aureus have resulted via exchange of mobile elements between these two species. Emergence of food-borne ST2250 S. argenteus is a potential threat to human public health. Highlights • Chickens at Chinese retail markets were screened for S. aureus including MRSA. • S. aureus (20.51%) and MRSA (1.58%) were detected in the investigated markets. • Antibiotic resistance remains a significant problem in poultry industry. • Emergence of foodborne ST2250 S. argenteus is a potential threat to public health. • Identical CRISPR types and spacers were detected in both S. argenteus and S. aureus. [ABSTRACT FROM AUTHOR]

[Display omitted] Antimicrobial resistance (AMR) is a global health threat. The dramatic increase of Methicillin-resistant Staphylococcus aureus (MRSA) infections emphasizes the need to find new anti-infective agents with a novel mode of action. The Caseinolytic protease (ClpP) is a central virulence factor in stress survival, virulence, and antibiotic resistance of MRSA. Here, we found ayanin, a flavonoid isolated from Callicarpa nudiflora , was an inhibitor of MRSA ClpP with an IC 50 of 19.63 μM. Using quantitative real-time PCR, ayanin reduced the virulence of Staphylococcus aureus (S. aureus) by down-regulating the level of some important virulence factors, including agrA , RNAⅢ , hla , pvl , psmα and spa. The results of cellular thermal shift assay and thermal shift assay revealed a binding between ayanin and ClpP. Molecular docking showed that ASP-168, ASN-173 and ARG-171 were the potential binding sites for ClpP binding to ayanin. ClpP mutagenesis study further indicated that ARG-171 and ASN-173 were the main active sites of ClpP. The affinity constant (K D) value of ayanin with ClpP was 3.15 × 10−5 M measured by surface plasmon resonance. In addition, ayanin exhibited a significant therapeutic effect on pneumonia infection induced by S. aureus in mice in vivo , especially in combination with vancomycin. This is the first report of ayanin with in vivo and in vitro efficacy against S. aureus infection. In conclusion, ayanin is a promising therapeutic agent to combat MRSA infections by targeting ClpP. [ABSTRACT FROM AUTHOR]

Avian arthritis is a relatively common disease in the poultry industry, the cause of which is complex. Bacterial arthritis is often caused by infection of Staphylococcus aureus ( S. aureus ), whereas viral arthritis is caused by avian orthoreovirus (ARV). To investigate the infection of S. aureus and ARV in cases of avian arthritis, a total of 77 samples characterized by arthritis were collected and detection. The results showed that 68.83% of the samples were positive for ARV, and 66.23% were positive for S. aureus. Among them, the ARV mono-infection rate was 22.08%, the S. aureus mono-infection rate was 19.48%, and ARV and S. aureus co-infection rate was 45.45%, indicating that ARV and S. aureus co-infection is common in arthritis cases. To further investigate the synergistic pathogenicity of ARV and S. aureus , ARV and S. aureus were used to mono-infect, co-infect, and (or) sequential infect SPF chickens and the clinical indications, pathologic changes, ARV load, S. aureus bacterial distribution, and cytokine level of the challenged chickens were evaluated. Decreased weight gain, increased mortality, and difficulties in standing were observed in all dual-infected groups and the singular-infected group. There were significantly more severe macroscopic and microscopic hock lesions, and larger amounts of a wider range of tissue distribution of ARV antigens and S. aureus bacterial distribution in the dual-infected groups compared to the single-infected and control groups. Cytokine detection showed a significant change in IFN-γ, IL-1β, and IL-6 levels in the infected groups, especially in the ARV– S. aureus co-infection, and (or) sequential infection groups, compared with the control group. Hence, ARV and S. aureus synergistically increased mortality in infected chickens, potentiated the severity of arthritis, and increased the amount of ARV RNA in tendons. [ABSTRACT FROM AUTHOR]

Toll-like receptor (TLR) signaling is an indispensable factor in immune cells activation. Many TLR ligands have been identified, and were characterized the immunological functions such as inflammatory cytokine production in immune cells. However, the anti-inflammatory response in TLR ligand-mediated manner is poorly understood. In this report, we show that bacterial lipoteichoic acid (LTA), which is a TLR2 ligand from gram-positive bacteria including Staphylococcus aureus (S. aureus), suppresses TLR-mediated inflammatory response in dendritic cells (DCs). The TLR ligand-induced Tumor Necrosis Factor-alpha (TNF-α) production was suppressed in the bone marrow derived dendritic cells (BMDCs) by co-treatment of LTA. The cellular activation, which was characterized as upregulations of CD80, CD86 and major histocompatibility complex II (MHC II) expression, was also suppressed in the TLR ligand stimulated BMDCs in the presence of LTA. While LTA itself didn’t induced both TNF-α production and upregulation of cell surface markers. The LTA mediated immunosuppressive function was abolished by TLR2 blocking in lipopolysaccharide (LPS)-stimulated BMDCs. Furthermore, LTA also showed the immunosuppressive function in the generation of IFN-γ+CD4+ T (Th1) cells by attenuation of antigen presenting activity in the BMDCs. In the imiquimod (IMQ)-induced acute skin inflammation, LTA suppressed the inflammation by downregulation of the activation in skin accumulated DCs. Thus, LTA is a TLR2 dependent immunological suppressor against inflammatory response induced by other TLR ligands in the DCs.

Osteomyelitis (OM) is a complicated and serious disease and its underlying molecular signatures of disease initiation and progression remain unclear. Staphylococcus aureus (S. aureus) is the most common causative agent of OM. Previous study of Banchereau et al. has established a link between whole blood transcription profiles and clinical manifestations in patients infected with S. aureus. However, the differentially expressed genes (DEGs) in OM induced by S. aureus infection have not been intensively investigated. In this study, we downloaded the gene expression profile dataset GSE30119 from Gene Expression Omnibus, and performed bioinformatic analysis to identify DEGs in S. aureus infection induced OM from the transcriptional level. The study consisted of 143 whole blood samples, including 44 healthy controls, 42 OM-free, and 57 OM infection patients. A total of 209 S. aureus infection-related genes (SARGs) and 377 OM-related genes (OMRGs) were identified. The SARGs were primarily involved in the immune response by GO functional and pathway enrichment analysis. Several proteins adhere to neutrophil extracellular traps may be critical for the immune response to the process of S. aureus infection. By contrast, the OMRGs differ from the SARGs. The OMRGs were enriched in transmembrane signaling receptor and calcium channel activity, cilium morphogenesis, chromatin silencing, even multicellular organism development. Several key proteins, including PHLPP2 and EGF, were hub nodes in protein–protein interaction network of the OMRGs. In addition, alcoholism, systemic lupus erythematosus and proteoglycans in cancer were the top pathways influenced by the OMRGs associated with OM. Thus, this study has further explored the DEGs and their biological functions associated with S. aureus infection and OM, comparing with the previous study, and may light the further insight into the underlying molecular mechanisms and the potential critical biomarkers in OM development.

Staphylococcus aureus (S. aureus) is a serious global pathogen that causes a diverse range of invasive diseases. S. aureus utilizes a family of pore-forming toxins, known as bi-component leukocidins, to evade the host immune response and promote infection. Among these is LukAB (leukocidin A/leukocidin B), a toxin that assembles into an octameric β-barrel pore in the target cell membrane, resulting in host cell death. The established cellular receptor for LukAB is CD11b of the Mac-1 complex. Here, we show that hydrogen voltage-gated channel 1 is also required for the cytotoxicity of all major LukAB variants. We demonstrate that while each receptor is sufficient to recruit LukAB to the plasma membrane, both receptors are required for maximal lytic activity. Why LukAB requires two receptors, and how each of these receptors contributes to pore-formation remains unknown. To begin to resolve this, we performed an alanine scanning mutagenesis screen to identify mutations that allow LukAB to maintain cytotoxicity without CD11b. We discovered 30 mutations primarily localized in the stem domains of LukA and LukB that enable LukAB to exhibit full cytotoxicity in the absence of CD11b. Using crosslinking, electron microscopy, and hydroxyl radical protein footprinting, we show these mutations increase the solvent accessibility of the stem domain, priming LukAB for oligomerization. Together, our data support a model in which CD11b binding unlatches the membrane penetrating stem domains of LukAB, and this change in flexibility promotes toxin oligomerization., Competing Interests: Conflict of interest V. J. T. is an inventor on patents and patent applications filed by New York University, which are currently under commercial license to Janssen Biotech Inc. Janssen Biotech provides research funding and other payments associated with the licensing agreement. All other authors declare no conflict of interest., (Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.)