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Ana Carolina Mazarin de Moraes,1 Bruna Araujo Lima,2 Andreia Fonseca de Faria,1 Marcelo Brocchi,2 Oswaldo Luiz Alves1 1Laboratory of Solid State Chemistry, Institute of Chemistry, University of Campinas, Campinas, São Paulo, Brazil; 2Department of Genetics, Evolution and Bioagents, Institute of Biology, University of Campinas, Campinas, São Paulo, Brazil Background: Methicillin-resistant Staphylococcus aureus (MRSA) has been responsible for serious hospital infections worldwide. Nanomaterials are an alternative to conventional antibiotic compounds, because bacteria are unlikely to develop microbial resistance against nanomaterials. In the past decade, graphene oxide (GO) has emerged as a material that is often used to support and stabilize silver nanoparticles (AgNPs) for the preparation of novel antibacterial nanocomposites. In this work, we report the synthesis of the graphene-oxide silver nanocomposite (GO-Ag) and its antibacterial activity against relevant microorganisms in medicine. Materials and methods: GO-Ag nanocomposite was synthesized through the reduction of silver ions (Ag+) by sodium citrate in an aqueous GO dispersion, and was extensively characterized using ultraviolet-visible absorption spectroscopy, X-ray diffraction, thermogravimetric analysis, X-ray photoelectron spectroscopy, and transmission electron microscopy. The antibacterial activity was evaluated by microdilution assays and time-kill experiments. The morphology of bacterial cells treated with GO-Ag was investigated via transmission electron microscopy. Results: AgNPs were well distributed throughout GO sheets, with an average size of 9.4±2.8 nm. The GO-Ag nanocomposite exhibited an excellent antibacterial activity against methicillin-resistant S. aureus, Acinetobacter baumannii, Enterococcus faecalis, and Escherichia coli. All (100%) MRSA cells were inactivated after 4 hours of exposure to GO-Ag sheets. In addition, no toxicity was found for either pristine GO or bare AgNPs within the tested concentration range. Transmission electronic microscopy images offered insights into how GO-Ag nanosheets interacted with bacterial cells. Conclusion: Our results indicate that the GO-Ag nanocomposite is a promising antibacterial agent against common nosocomial bacteria, particularly antibiotic-resistant MRSA. Morphological injuries on MRSA cells revealed a likely loss of viability as a result of the direct contact between bacteria and the GO-Ag sheets. Keywords: graphene oxide, silver nanoparticles, graphene oxide-silver nanocomposite, antibacterial agent, MRSA, Escherichia coli

Abstract: Morpho‐physiological changes caused by particular plant growth‐promoting rhizobacteria were evaluated in sugarcane plants under varying water availability. Under well‐watered conditions, we have found one rhizobacteria isolate (IAC‐RBcr5) able to enhance root dry matter and photosynthesis of sugarcane plants. The IAC‐RBcr5 genome was sequenced and high similarity was found with Pseudomonas putida GB‐1. Based on increased root system size of inoculated plants, we hypothesised that sugarcane plants inoculated with IAC‐RBcr5 would have improved performance under water deficit. Although IAC‐RBcr5 had improved plant leaf CO2 assimilation under water shortage, inoculation caused reduction of biomass accumulation in sugarcane. The negative influence of water deficit on shoot growth rate and root traits such as volume, area, diameter, length and specific root area was higher in plants treated with IAC‐RBcr5 as compared to non‐inoculated ones. However, rhizobacteria‐induced improvements in leaf and root proline contents would represent a strategy for storing carbon and nitrogen during low water availability and helping both organisms to resume their metabolism after rehydration. In conclusion, we found and identified a rhizobacterium able to improve growth and photosynthesis of sugarcane plants. Such benefit for plant growth was lost under low water availability as a likely consequence of increased carbon‐energy demand by rhizobacteria and their sensitivity to drought. [ABSTRACT FROM AUTHOR]

Since violacein-an antibiotic, antiviral, and antiparasitic compound-exhibits poor solubility in water, polymeric poly-( d, l-lactide-co-glycolide) nanoparticles containing this compound improved its solubility and biological activity. The nanoparticles were prepared by the nanoprecipitation method and characterized in terms of average diameter, zeta potential, drug loading, polymer recovery, in vitro release kinetic, and in vitro antibacterial activity. Nanoparticles with diameters between 116 and 139 nm and negative-charged outer surfaces were obtained. Drug-loading efficiency and polymer recovery were 87 and 93%, respectively. In vitro release kinetics assays showed that violacein loaded in these nanoparticles has sustained release behavior until 5 days. Both free and nanoparticles-loaded violacein exhibited in vitro antibacterial activity against Staphylococcus aureus ATCC 29213 and ATCC 25923 strains and exhibiting around two to five times lower minimum inhibitory concentration (MIC) than free violacein, respectively. The encapsulated violacein was efficient against methicilin-resistant Staphylococcus aureus (MRSA) strains. No significant activity against Escherichia coli and Salmonella enterica was found. [ABSTRACT FROM AUTHOR]

Abstract Background All organisms living under aerobic atmosphere have powerful mechanisms that confer their macromolecules protection against oxygen reactive species. Microorganisms have developed biomolecule-protecting systems in response to starvation and/or oxidative stress, such as DNA biocrystallization with Dps (DNA-binding protein from starved cells). Dps is a protein that is produced in large amounts when the bacterial cell faces harm, which results in DNA protection. In this work, we evaluated the glycosylation in the Dps extracted from Salmonella enterica serovar Typhimurium. This Dps was purified from the crude extract as an 18-kDa protein, by means of affinity chromatography on an immobilized jacalin column. Results The N-terminal sequencing of the jacalin-bound protein revealed 100% identity with the Dps of S. enterica serovar Typhimurium. Methyl-alpha-galactopyranoside inhibited the binding of Dps to jacalin in an enzyme-linked lectin assay, suggesting that the carbohydrate recognition domain (CRD) of jacalin is involved in the interaction with Dps. Furthermore, monosaccharide compositional analysis showed that Dps contained mannose, glucose, and an unknown sugar residue. Finally, jacalin-binding Dps was detected in larger amounts during the bacterial earlier growth periods, whereas high detection of total Dps was verified throughout the bacterial growth period. Conclusion Taken together, these results indicate that Dps undergoes post-translational modifications in the pre- and early stationary phases of bacterial growth. There is also evidence that a small mannose-containing oligosaccharide is linked to this bacterial protein.

Background: To prospectively evaluate ultrasound backscatter coefficients (BSCs) of the supraspinatus and infraspinatus muscles and compare with Goutallier classification on magnetic resonance imaging (MRI)., Methods: Fifty-six participants had shoulder MRI exams and ultrasound exams of the supraspinatus and infraspinatus muscles. Goutallier MRI grades were determined and BSCs were measured. Group means were compared and the strength of relationships between the measures were determined. Using binarized Goutallier groups (0-2 versus 3-4), areas under the receiver operating characteristic curves (AUROCs) were calculated. The nearest integer cutoff value was determined using Youden's index., Results: BSC values were significantly different among most Goutallier grades for the supraspinatus and infraspinatus muscles (both p < 0.001). Strong correlations were found between the BSC values and Goutallier grades for the supraspinatus (τ b  = 0.72, p < 0.001) and infraspinatus (τ b  = 0.79, p < 0.001) muscles. BSC showed excellent performance for classification of the binarized groups (0-2 versus 3-4) for both supraspinatus (AUROC = 0.98, p < 0.0001) and infraspinatus (AUROC = 0.98, p < 0.0001) muscles. Using a cutoff BSC value of -17 dB, sensitivity, specificity, and accuracy for severe fatty infiltration were 87.0%, 90.0%, and 87.5% for the supraspinatus muscle, and 93.6%, 87.5%, and 92.7% for the infraspinatus muscle., Conclusion: BSC can be applied to the rotator cuff muscles for assessment of fatty infiltration. For both the supraspinatus and infraspinatus muscles, BSC values significantly increased with higher Goutallier grades and showed strong performance in distinguishing low versus high Goutallier grades., Relevance Statement: Fatty infiltration of the rotator cuff muscles can be quantified using BSC values, which are higher with increasing Goutallier grades., Key Points: Ultrasound BSC measurements are reliable for the quantification of muscle fatty infiltration. BCS values increased with higher Goutallier MRI grades. BCS values demonstrated high performance for distinguishing muscle fatty infiltration groups., (© 2024. This is a U.S. Government work and not under copyright protection in the US; foreign copyright protection may apply.)

Diffusion tensor imaging (DTI) is a magnetic resonance imaging (MRI) technique that can be used to assess microstructural features of skeletal muscle that are related to tissue function. Although widely used, direct correlations between DTI derived metrics such as fractional anisotropy and spatially matched tissue microstructure assessed with histology have not been performed. This study investigated the relationship between scalar-based DTI measurements and histologically derived muscle microstructural measurements in rat rotator cuff muscles. Despite meticulous co-localization of MRI and histology data, negligible correlations were found between DTI metrics and histological measurements including muscle fiber diameter, cross-sectional area, and surface-to-volume ratio. These findings highlight the challenges in validating DTI with histology due to requirements in anatomical co-localization, necessity of high-quality histology, and consideration of diffusion measurement scales. Our findings underscore the need for further research with optimized imaging parameters to enhance our knowledge regarding the sensitivity of DTI to important features of muscle microstructure., Competing Interests: Competing Interests Statement The authors declare no competing interests.

The lack of effective treatment options for an increasing number of cancer cases highlights the need for new anticancer therapeutic strategies. Immunotherapy mediated by Salmonella enterica Typhimurium is a promising anticancer treatment. Candidate strains for anticancer therapy must be attenuated while retaining their antitumor activity. Here, we investigated the attenuation and antitumor efficacy of two S. enterica Typhimurium mutants, Δ tolRA and Δ ihfABpmi , in a murine melanoma model. Results showed high attenuation of Δ tolRA in the Galleria mellonella model, and invasion and survival in tumor cells. However, it showed weak antitumor effects in vitro and in vivo . Contrastingly, lower attenuation of the attenuated Δ ihfABpmi strain resulted in regression of tumor mass in all mice, approximately 6 days after the first treatment. The therapeutic response induced by Δ ihfABpmi was accompanied with macrophage accumulation of antitumor phenotype (M1) and significant increase in the mRNAs of proinflammatory mediators (TNF-α, IL-6, and iNOS) and an apoptosis inducer (Bax). Our findings indicate that the attenuated Δ ihfABpmi exerts its antitumor activity by inducing macrophage infiltration or reprogramming the immunosuppressed tumor microenvironment to an activated state, suggesting that attenuated S. enterica Typhimurium strains based on nucleoid-associated protein genes deletion could be immunotherapeutic against cancer., 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 Pérez Jorge, Gontijo, Silva, Goes, Jaimes-Florez, Coser, Rocha, Giorgio and Brocchi.)

Endolysins are bacteriophage (or phage)-encoded enzymes that catalyse the peptidoglycan breakdown in the bacterial cell wall. The exogenous action of recombinant phage endolysins against Gram-positive organisms has been extensively studied. However, the outer membrane acts as a physical barrier when considering the use of recombinant endolysins to combat Gram-negative bacteria. This study aimed to evaluate the antimicrobial activity of the SAR-endolysin LysKpV475 against Gram-negative bacteria as single or combined therapies, using an outer membrane permeabilizer (polymyxin B) and a phage, free or immobilized in a pullulan matrix. In the first step, the endolysin LysKpV475 in solution, alone and combined with polymyxin B, was tested in vitro and in vivo against ten Gram-negative bacteria, including highly virulent strains and multidrug-resistant isolates. In the second step, the lyophilized LysKpV475 endolysin was combined with the phage phSE-5 and investigated, free or immobilized in a pullulan matrix, against Salmonella enterica subsp. enterica serovar Typhimurium ATCC 13311. The bacteriostatic action of purified LysKpV475 varied between 8.125 μg ml -1 against Pseudomonas aeruginosa ATCC 27853, 16.25 μg ml -1 against S . enterica Typhimurium ATCC 13311, and 32.50 μg ml -1 against Klebsiella pneumoniae ATCC BAA-2146 and Enterobacter cloacae P2224. LysKpV475 showed bactericidal activity only for P. aeruginosa ATCC 27853 (32.50 μg ml -1 ) and P. aeruginosa P2307 (65.00 μg ml -1 ) at the tested concentrations. The effect of the LysKpV475 combined with polymyxin B increased against K. pneumoniae ATCC BAA-2146 [fractional inhibitory concentration index (FICI) 0.34; a value lower than 1.0 indicates an additive/combined effect] and S . enterica Typhimurium ATCC 13311 (FICI 0.93). A synergistic effect against S . enterica Typhimurium was also observed when the lyophilized LysKpV475 at ⅔ MIC was combined with the phage phSE-5 (m.o.i. of 100). The lyophilized LysKpV475 immobilized in a pullulan matrix maintained a significant S almonella reduction of 2 logs after 6 h of treatment. These results demonstrate the potential of SAR-endolysins, alone or in combination with other treatments, in the free form or immobilized in solid matrices, which paves the way for their application in different areas, such as in biocontrol at the food processing stage, biosanitation of food contact surfaces and biopreservation of processed food in active food packing.

Purpose: Creating an effective MRI protocol for examining the brachial plexus poses significant challenges, and despite the abundance of protocols in the literature, there is a lack of reference standards for basic sequences and essential parameters needed for replication. The aim of this study is to establish a reproducible 1.5 T brachial plexus imaging protocol, including patient positioning, coil selection, imaging planes, and essential sequence parameters., Methods: We systematically investigated MRI sequences, testing each parameter through in vivo experiments, examining their effects on signal-to-noise ratio (SNR), contrast-to-noise ratio (CNR), visual quality scores, and acquisition time. Sequences were refined based on optimal quality and timing scores. The final protocol was tested on scanners from two other vendors for reliability., Results: The final protocol included a combination of 2D turbo-spin-echo and 3D SPACE T1, SPACE STIR, and VIBE DIXON sequences. Recommendations for imaging planes, phase encoding, field of view, TR, TE, resolution, number of slices, slice thickness, fat and blood suppression, and acceleration strategies are provided. The protocol was successfully translated to other vendor's scanners with comparable quality., Conclusion: We present an optimized protocol detailing the essential parameters for reproducibility. Our comprehensive list of experiments describes the impact of each parameter on image quality and scan time, addressing common artifacts and potential solutions. This protocol can benefit both young radiologists new to the field and experienced professionals seeking to refine their existing protocols., Competing Interests: Declaration of conflicting interestsThe author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

Conventional cancer therapies have many limitations. In the last decade, it has been suggested that bacteria-mediated immunotherapy may circumvent the restrictions of traditional treatments. For example, Salmonella enterica is the most promising bacteria for treating cancer due to its intrinsic abilities, such as killing tumor cells, targeting, penetrating, and proliferating into the tumor. S. enterica has been genetically modified to ensure safety and increase its intrinsic antitumor efficacy. This bacterium has been used as a vector for delivering anticancer agents and as a combination therapy with chemotherapy, radiotherapy, or photothermic. Recent studies have reported the antitumor efficacy of outer membrane vesicles (OMVs) derived from S. enterica . OMVs are considered safer than attenuated bacteria and can stimulate the immune system as they comprise most of the immunogens found on the surface of their parent bacteria. Furthermore, OMVs can also be used as nanocarriers for antitumor agents. This review describes the advances in S. enterica as immunotherapy against cancer and the mechanisms by which Salmonella fights cancer. We also highlight the use of OMVs as immunotherapy and nanocarriers of anticancer agents. OMVs derived from S. enterica are innovative and promising strategies requiring further investigation., 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 © 2023 Pérez Jorge, Gontijo and Brocchi.)

Introduction: Obstruction of the lacrimal drainage represents a common ophthalmologic issue. The blockage may interest any level of the lacrimal drainage pathway, and it is important to find the site of obstruction to plan the most appropriate treatment. In this study, findings from magnetic resonance (MR) dacryocystography were compared with findings from endoscopic and surgical procedures to evaluate the accuracy of MR dacryocystography in localizing the site of nasolacrimal duct obstruction., Methods: We enrolled twenty-one patients with clinical suspicion of nasolacrimal duct obstruction who underwent dacryoendoscopy and surgery. MR dacryocystography was performed with a heavily T2-weighted fast spin echo sequence in the coronal planes. Before the MRI was performed, a sterile 0.9% NaCl solution was administered into both conjunctival sacs. For each examination, two independent readers (with 8 and 10 years of experience in head and neck imaging) evaluated both heavily 3D space T2-weighted and STIR sequences., Results: Stenosis/obstruction of nasolacrimal duct or lacrimal sac was diagnosed in all 21 patients who underwent MRI dacryocystography. In particular, the site of the obstruction was classified as lacrimal sac in 12 (57%) patients, nasolacrimal duct in 6 (29%) patients, and canaliculi in 3 (14%) patients by both readers. By comparison with the evidence resulting from the endoscopy, there were differences between MRI dacryocystography and dacryoendoscopy in the evaluation of the obstruction's site in three patients, with an overall accuracy of 85.7%., Conclusion: MR dacryocystography allows a non-invasive evaluation of the lacrimal drainage pathway, valid for the planning of the most appropriate treatment.

The emergence and re-emergence of bacterial strains resistant to multiple drugs represent a global health threat, and the search for novel biological targets is a worldwide concern. AhpC are enzymes involved in bacterial redox homeostasis by metabolizing diverse kinds of hydroperoxides. In pathogenic bacteria, AhpC are related to several functions, as some isoforms are characterized as virulence factors. However, no inhibitor has been systematically evaluated to date. Here we show that the natural ent -kaurane Adenanthin (Adn) efficiently inhibits AhpC and molecular interactions were explored by computer assisted simulations. Additionally, Adn interferes with growth and potentializes the effect of antibiotics (kanamycin and PMBN), positioning Adn as a promising compound to treat infections caused by multiresistant bacterial strains.