Phage-antibiotic combinations in various treatment modalities to manage MRSA infections.

Introduction: The emergence of antibiotic resistance is a significant challenge in the treatment of bacterial infections, particularly in patients in the intensive care unit (ICU). Phage-antibiotic combination therapy is now being utilized as a preferred therapeutic option for infections that are multi-drug resistant in nature. Methods: In this study, we examined the combined impact of the staph phage vB_Sau_S90 and four antibiotics on methicillin-resistant Staphylococcus aureus (MRSA). We conducted experiments on three different treatment sequences: a) administering phages before antibiotics, b) administering phages and antibiotics simultaneously, and c) administering antibiotics before phages. Results: When the media was supplemented with sub-inhibitory concentrations of 0.25 µg/mL and 1 µg/mL, the size of the plaque increased from 0.5 ± 0.1 mm (in the control group with only the phage) to 4 ± 0.2 mm, 1.6 ± 0.1 mm, and 1.6 ± 0.4 mm when fosfomycin, ciprofloxacin, and oxacillin were added, respectively. The checkerboard analysis revealed a synergistic effect between the phages and antibiotics investigated, as indicated by a FIC value of less than 0.5. The combination treatment of phages and antibiotics demonstrated universal efficacy across all treatments. Nevertheless, the optimal effectiveness was demonstrated when the antibiotics were delivered subsequent to the phages. Utilizing the Galleria mellonella model, in vivo experiments showed that the combination of phage-oxacillin effectively eliminated biofilm-infected larvae, resulting in a survival rate of up to 80% in the treated groups. Discussion: Our findings highlight the advantages of using a combination of phage and antibiotic over using phages alone in the treatment of MRSA infections.

Complete Genome Sequence of Pseudomonas Phage Motto.

We describe the complete genome sequence of bacteriophage Motto, which infects clinical strains of Pseudomonas aeruginosa. Motto is a T1-like siphovirus related to members of the family Drexlerviridae and has a capsid width of ~57 nm and a tail length of ~255 nm. The 49.9-kb genome contains 84 protein-coding genes.

A Multiwell-Plate Caenorhabditis elegans Assay for Assessing the Therapeutic Potential of Bacteriophages against Clinical Pathogens.

In order to establish phage therapy as a standard clinical treatment for bacterial infections, testing of every phage to ensure the suitability and safety of the biological compound is required. While some issues have been addressed over recent years, standard and easy-to-use animal models to test phages are still rare. Testing of phages in highly suitable mammalian models such as mice is subjected to strict ethical regulations, while insect larvae such as the Galleria mellonella model suffer from batch-to-batch variations and require manual operator skills to inject bacteria, resulting in unreliable experimental outcomes. A much simpler model is the nematode Caenorhabditis elegans, which feeds on bacteria, a fast growing and easy to handle organism that can be used in high-throughput screening. In this study, two clinical bacterial strains of Escherichia coli, one Klebsiella pneumoniae, and one Enterobacter cloacae strain were tested on the model system together with lytic bacteriophages that we isolated previously. We developed a liquid-based assay, in which the efficiency of phage treatment was evaluated using a scoring system based on microscopy and counting of the nematodes, allowing increasing statistical significance compared to other assays such as larvae or mice. Our work demonstrates the potential to use Caenorhabditis elegans to test the virulence of strains of Klebsiella pneumoniae, Enterobacter cloacae, and EHEC/EPEC as well as the efficacy of bacteriophages to treat or prevent infections, allowing a more reliable evaluation for the clinical therapeutic potential of lytic phages. IMPORTANCE Validating the efficacy and safety of phages prior to clinical application is crucial to see phage therapy in practice. Current animal models include mice and insect larvae, which pose ethical or technical challenges. This study examined the use of the nematode model organism C. elegans as a quick, reliable, and simple alternative for testing phages. The data show that all the four tested bacteriophages can eliminate bacterial pathogens and protect the nematode from infections. Survival rates of the nematodes increased from <20% in the infection group to >90% in the phage treatment group. Even the nematodes with poly-microbial infections recovered during phage cocktail treatment. The use of C. elegans as a simple whole-animal infection model is a rapid and robust way to study the efficacy of phages before testing them on more complex model animals such as mice.

Antibiotic resistance, biofilm forming ability, and clonal profiling of clinical isolates of Staphylococcus aureus from southern and northeastern India.

Background: Staphylococcus aureus is a pathogen endemic in India and sometimes deadly for patients in intensive care units. Objectives: To determine the antibiotic-resistance pattern, biofilm forming ability, and clonal type of S. aureus from isolates collected in Tamil Nadu (south) and the Mizoram (northeast) regions of India. Methods: We collected S. aureus isolates from diagnostic laboratories in Tamil Nadu and Mizoram. An antibiotic susceptibility test was performed according to Clinical Laboratory and Standards Institute methods. Antibiotic-resistant determinants such as mecA, mecC, blaZ, vanA, vanB, and vanC were confirmed by polymerase chain reaction (PCR). All isolates were further studied for biofilm forming ability. Enterobacterial repetitive intergenic consensus (ERIC)-PCR was used for clonal analysis. Results: A study of 206 clinical isolates showed 52.9% prevalence of methicillin-resistant S. aureus in Tamil Nadu and 49.4% in Mizoram. Minimum inhibitory concentration tests showed a high prevalence of 67% oxacillin resistance in isolates from Tamil Nadu and 49% in isolates from Mizoram. PCR showed 53% mecA in Tamil Nadu and 49% mecA in Mizoram. Vancomycin-intermediate resistance S. aureus (VISA) prevalence was lower in isolates from Tamil Nadu (4%) and Mizoram (5%). All methicillin-resistant S. aureus (MRSA) isolates formed biofilms. Clonal analysis revealed a genetic relatedness between the isolates. Conclusions: The prevalence of MRSA is high in the regions studied, with most of the clinical isolates being multidrug resistant. Adopting appropriate community-based preventive measures and establishing antimicrobial stewardship is highly recommended to minimize the dissemination in antibiotic resistance.

Phage therapy as a revolutionary medicine against Gram-positive bacterial infections.

BACKGROUND: Antibiotic resistance among pathogenic bacteria has created a global emergency, prompting the hunt for an alternative cure. Bacteriophages were discovered over a century ago and have proven to be a successful replacement during antibiotic treatment failure. This review discusses on the scientific investigation of phage therapy for Gram-positive pathogens and general outlook of phage therapy clinical trials and commercialization. MAIN BODY OF THE ABSTRACT: This review aimed to highlight the phage therapy in Gram-positive bacteria and the need for phage therapy in the future. Phage therapy to treat Gram-positive bacterial infections is in use for a very long time. However, limited review on the phage efficacy in Gram-positive bacteria exists. The natural efficiency and potency of bacteriophages against bacterial strains have been advantageous amidst the other non-antibiotic agents. The use of phages to treat oral biofilm, skin infection, and recurrent infections caused by Gram-positive bacteria has emerged as a predominant research area in recent years. In addition, the upsurge in research in the area of phage therapy for spore-forming Gram-positive bacteria has added a wealth of information to phage therapy. SHORT CONCLUSION: We conclude that the need of phage as an alternative treatment is obvious in future. However, phage therapy can be used as reserve treatment. This review focuses on the potential use of phage therapy in treating Gram-positive bacterial infections, as well as their therapeutic aspects. Furthermore, we discussed the difficulties in commercializing phage drugs and their problems as a breakthrough medicine.

A Method to Quantify Tensile Biaxial Properties of Mouse Aortic Valve Leaflets.

Understanding aortic valve (AV) mechanics is crucial in elucidating both the mechanisms that drive the manifestation of valvular diseases as well as the development of treatment modalities that target these processes. Genetically modified mouse models have become the gold standard in assessing biological mechanistic influences of AV development and disease. However, very little is known about mouse aortic valve leaflet (MAVL) tensile properties due to their microscopic size (∼500 µm long and 45 µm thick) and the lack of proper mechanical testing modalities to assess uniaxial and biaxial tensile properties of the tissue. We developed a method in which the biaxial tensile properties of MAVL tissues can be assessed by adhering the tissues to a silicone rubber membrane utilizing dopamine as an adhesive. Applying equiaxial tensile loads on the tissue-membrane composite and tracking the engineering strains on the surface of the tissue resulted in the characteristic orthotropic response of AV tissues seen in human and porcine tissues. Our data suggest that the circumferential direction is stiffer than the radial direction (n = 6, P = 0.0006) in MAVL tissues. This method can be implemented in future studies involving longitudinal mechanical stimulation of genetically modified MAVL tissues bridging the gap between cellular biological mechanisms and valve mechanics in popular mouse models of valve disease.

Uncovering the Mechanical, Thermal, and Chemical Characteristics of Biodegradable Mushroom Leather with Intrinsic Antifungal and Antibacterial Properties.

The growing demand for a sustainable leather industry with a low environmental impact has prompted the development of alternative vegetable-based materials. In this study, a biodegradable mushroom-based leather derived from the fruiting body of Phellinus ellipsoideus is investigated. The biodegradable leather proves to be thermally stable up to 250 °C. The mechanically robust macrostructure combines a tensile strength of 1.2 MPa and ductility (101% strain at break) attributed to the natural balance of chitin (0.3) and proteins (0.7) constituting the mycelium fibers. The chitin-protein system results in an intrinsic scratch-resistant structure with exciting damping properties in a low frequency range. Enhanced damping capabilities within 5-20 Hz (tan δ: 0.1-0.20) are attributed to the macrostuctural alignment of the mycelium under cyclic tension. Whereas, increasing frequencies >20 Hz induce micromolecular interactions between chitin and proteins within the fibers. Exposure of the bioleather to acidic (pH 4, 5) and basic (pH 8, 9) media demonstrated the selective dissolution of proteins (basic) and chitin (acid) components within the mycelium, opening an opportunity for tunable mechanical response. Reducing the protein content induced an increase in stiffness and strength (pH 8 and 9), while reducing its chitin component showed variable ductility (pH 4 and 5). Owing to the entirely natural composition of the mushroom leather, intrinsic antifungal and antibacterial properties found in the mycelium resist fungal invasion and bacterial growth. Thus, this study displays the unique morphology-property relationship of a biodegradable mushroom leather, proving its potential as a fully sustainable and environmentally friendly alternative.

Quantification of Thermal Oxidation in Metallic Glass Powder using Ultra-small Angle X-ray Scattering.

In this paper, the composition, structure, morphology and kinetics of evolution during isothermal oxidation of Fe48Cr15Mo14Y2C15B6 metallic glass powder in the supercooled region are investigated by an integrated ex-situ and in-situ characterization and modelling approach. Raman and X-ray diffraction spectra established that oxidation yielded a hierarchical structure across decreasing length scales. At larger scale, Fe2O3 grows as a uniform shell over the powder core. This shell, at smaller scale, consists of multiple grains. Ultra-small angle X-ray scattering intensity acquired during isothermal oxidation of the powder over a wide Q-range delineated direct quantification of oxidation behavior. The hierarchical structure was employed to construct a scattering model that was fitted to the measured intensity distributions to estimate the thickness of the oxide shell. The relative gain in mass during oxidation, computed theoretically from this model, relatively underestimated that measured in practice by a thermogravimetric analyzer due to the distribution in sizes of the particles. Overall, this paper presents the first direct quantification of oxidation in metallic glass powder by ultra-small angle X-ray scattering. It establishes novel experimental environments that can potentially unfold new paradigms of research into a wide spectrum of interfacial reactions in powder materials at elevated temperatures.

Three-Dimensional Graphene Foam Induces Multifunctionality in Epoxy Nanocomposites by Simultaneous Improvement in Mechanical, Thermal, and Electrical Properties.

Three-dimensional (3D) macroporous graphene foam based multifunctional epoxy composites are developed in this study. Facile dip-coating and mold-casting techniques are employed to engineer microstructures with tailorable thermal, mechanical, and electrical properties. These processing techniques allow capillarity-induced equilibrium filling of graphene foam branches, creating epoxy/graphene interfaces with minimal separation. Addition of 2 wt % graphene foam enhances the glass transition temperature of epoxy from 106 to 162 °C, improving the thermal stability of the polymer composite. Graphene foam aids in load-bearing, increasing the ultimate tensile strength by 12% by merely 0.13 wt % graphene foam in an epoxy matrix. Digital image correlation (DIC) analysis revealed that the graphene foam cells restrict and confine the deformation of the polymer matrix, thereby enhancing the load-bearing capability of the composite. Addition of 0.6 wt % graphene foam also enhances the flexural strength of the pure epoxy by 10%. A 3D network of graphene branches is found to suppress and deflect the cracks, arresting mechanical failure. Dynamic mechanical analysis (DMA) of the composites demonstrated their vibration damping capability, as the loss tangent (tan δ) jumps from 0.1 for the pure epoxy to 0.24 for ∼2 wt % graphene foam-epoxy composite. Graphene foam branches also provide seamless pathways for electron transfer, which induces electrical conductivity exceeding 450 S/m in an otherwise insulator epoxy matrix. The epoxy-graphene foam composite exhibits a gauge factor as high as 4.1, which is twice the typical gauge factor for the most common metals. Simultaneous improvement in thermal, mechanical, and electrical properties of epoxy due to 3D graphene foam makes epoxy-graphene foam composite a promising lightweight and multifunctional material for aiding load-bearing, electrical transport, and motion sensing in aerospace, automotive, robotics, and smart device structures.

Oxidative Unzipping and Transformation of High Aspect Ratio Boron Nitride Nanotubes into "White Graphene Oxide" Platelets.

Morphological and chemical transformations in boron nitride nanotubes under high temperature atmospheric conditions is probed in this study. We report atmospheric oxygen induced cleavage of boron nitride nanotubes at temperatures exceeding 750 °C for the first time. Unzipping is then followed by coalescence of these densely clustered multiple uncurled ribbons to form stacks of 2D sheets. FTIR and EDS analysis suggest these 2D platelets to be Boron Nitride Oxide platelets, with analogous structure to Graphene Oxide, and therefore we term them as "White Graphene Oxide" (WGO). However, not all BNNTs deteriorate even at temperatures as high as 1000 °C. This leads to the formation of a hybrid nanomaterial system comprising of 1D BN nanotubes and 2D BN oxide platelets, potentially having advanced high temperature sensing, radiation shielding, mechanical strengthening, electron emission and thermal management applications due to synergistic improvement of multi-plane transport and mechanical properties. This is the first report on transformation of BNNT bundles to a continuous array of White Graphene Oxide nanoplatelet stacks.