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Escherichia coli (E. coli) detection suffers from slow analysis time and high costs, along with the need for specificity. While state-of-the-art electrochemical biosensors are cost-efficient and easy to implement, their sensitivity and analysis time still require improvement. In this work, we present a paper-based electrochemical biosensor utilizing magnetic core-shell Fe 2 O 3 @CdSe/ZnS quantum dots (MQDs) to achieve fast detection, low cost, and high sensitivity. Using electrochemical impedance spectroscopy (EIS) as the detection technique, the biosensor achieved a limit of detection of 2.7 × 10 2 CFU/mL for E. coli bacteria across a concentration range of 10 2 -10 8 CFU/mL, with a relative standard deviation (RSD) of 3.5781%. From an optical perspective, as E. coli concentration increased steadily from 10 4 to 10 7 CFU/mL, quantum dot fluorescence showed over 60% lifetime quenching. This hybrid biosensor thus provides rapid, highly sensitive E. coli detection with a fast analysis time of 30 min. This study, which combines the detection advantages of electrochemical and optical biosensor systems in a graphite-based paper sensor for the first time, has the potential to meet the needs of point-of-care applications. It is thought that future studies that will aim to examine the performance of the production-optimized, portable, graphite-based sensor system on real food samples, environmental samples, and especially medical clinical samples will be promising.

Antibiotic resistance is a global public health concern. Bacteria have evolved resistance to most antibiotics, which means that for any given bacterial infection, the bacteria may be resistant to one or several antibiotics. It has been suggested that genomic sequencing and machine learning (ML) could make resistance testing more accurate and cost-effective. Given that ML is likely to become an ever more important tool in medicine, we believe that it is important for pre-health students and others in the life sciences to learn to use ML tools. This paper provides a step-by-step tutorial to train 4 different ML models (logistic regression, random forests, extreme gradient-boosted trees, and neural networks) to predict drug resistance for Escherichia coli isolates and to evaluate their performance using different metrics and cross-validation techniques. We also guide the user in how to load and prepare the data used for the ML models. The tutorial is accessible to beginners and does not require any software to be installed as it is based on Google Colab notebooks and provides a basic understanding of the different ML models. The tutorial can be used in undergraduate and graduate classes for students in Biology, Public Health, Computer Science, or related fields., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Orcales et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

In recent years, whole-cell biosensors (WCBs) have emerged as a potent approach for environmental monitoring and on-site analyte detection. These biosensors harness the biological apparatus of microorganisms to identify specific analytes, offering advantages in sensitivity, specificity, and real-time monitoring capabilities. A critical hurdle in biosensor development lies in ensuring the robust attachment of cells to surfaces, a crucial step for practical utility. In this study, we present a comprehensive approach to tackle this challenge via engineering Escherichia coli cells for immobilization on paper through the Curli biofilm pathway. Furthermore, incorporating a cellulose-binding peptide domain to the CsgA biofilm protein enhances cell adhesion to paper surfaces, consequently boosting biosensor efficacy. To demonstrate the versatility of this platform, we developed a WCB for copper, optimized to exhibit a discernible response, even with the naked eye. To confirm its suitability for practical field use, we characterized our copper sensor under various environmental conditions-temperature, salinity, and pH-to mimic real-world scenarios. The biosensor-equipped paper discs can be freeze-dried for deployment in on-site applications, providing a practical method for long-term storage without loss of sensitivity paper discs demonstrate sustained functionality and viability even after months of storage with 5 μM limit of detection for copper with visible-to-naked-eye signal levels. Biofilm-mediated surface attachment and analyte sensing can be independently engineered, allowing for flexible utilization of this platform as required. With the implementation of copper sensing as a proof-of-concept study, we underscore the potential of WCBs as a promising avenue for the on-site detection of a multitude of analytes., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Urartu Ozgur Safak Seker reports financial support was provided by "The Scientific and Technological Research Council of Turkey". If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

This study introduces a novel paper coating approach using modified zinc oxide (ZnO), providing a comparison with conventional materials used in the paper industry. The research focused on determining the concentration for effective microbial growth inhibition and evaluates the impact of different ZnO types on coated-paper properties, including antimicrobial activity, surface morphology, tensile strength, and water absorption. Specifically, ZnO microparticles (ZnOws), ZnO nanoparticles (ZnOnp), and modified ZnOnp (ZnOnp-CaCO3, with a core–shell structure composed of calcium carbonate [CaCO3] and nano-zinc oxide) were incorporated into coating formulations at varying concentrations (0 × MIC, 1 × MIC, 2 × MIC, and 3 × MIC, based on minimum inhibitory concentrations [MICs]). The results demonstrated that among all tested microorganisms, ZnOnp-CaCO3 showed the lowest MIC values. ZnOnp-CaCO3-coated paper exhibited superior antimicrobial activity against both Gram-positive and Gram-negative bacteria, as well as fungi, outperforming ZnOws and ZnOnp. At 1 × MIC, %inhibition for E. coli, S. aureus, and A. niger were 98.3%, 99.1%, and 90.8%, respectively. Additionally, ZnOnp-CaCO3 coatings caused minimal color change in the paper compared to the other ZnO variants. The coating did not negatively impact the mechanical properties of the paper across all ZnO types and concentrations. Water absorption tests showed increased hydrophobicity with higher ZnO content, with ZnOnp and ZnOnp-CaCO3 exhibiting greater reductions in water absorption than ZnOws. Overall, ZnOnp-CaCO3 showed strong potential as an antimicrobial agent for paper surfaces, making it ideal for packaging and hygiene products. By partially replacing ZnOnp with inexpensive CaCO3 core particles, ZnOnp-CaCO3 delivers enhanced performance, reduced costs, and greater sustainability for large-scale applications. [ABSTRACT FROM AUTHOR]

In order to improve the paper properties and enhance the utilization efficiency of functional chemical additives, surface sizing agents were prepared by combining water-soluble maleic anhydride acylated chitosan (MAAC), carboxymethylated chitosan (O-CMCS) and polyvinyl alcohol (PVA), and the effects of sizing process on the surface morphology, physical strength and antimicrobial properties of the dialysis paper were investigated. The results showed that when the coating weight was about 6.5 g/m2 and the dosage of the modified chitosan was about 1.5% of the paper weight, the dry tensile strength, wet tensile strength and burst strength of paper sized with MAAC/PVA sizing agent increased by 46.5%, 82.2% and 107%, respectively, compared to that of unsized paper, while the dry tensile strength, wet tensile strength and burst strength of paper sized with O-CMCS/PVA sizing agent increased by 41.4%, 53.6% and 107%, respectively. In addition, the paper sized with MAAC/PVA and O-CMCS/PVA sizing agent displayed excellent antibacterial effect, which had the antibacterial rate against E. coli of 94.8% and 92.9%, respectively. In a word, the physical strength and antimicrobial properties of paper could be greatly improved by adding a small amount of modified chitosan (CS) to the surface sizing agent, and the strength improvement and antibacterial effect of MAAC/PVA sizing agent were better than that of O-CMCS/PVA. [ABSTRACT FROM AUTHOR]

Non-recyclable fiber rejects from paper mills, particularly those from recycled linerboard mills, contain high levels of structural carbohydrates but are currently landfilled, causing financial and environmental burdens. The aim of this study was to develop efficient and sustainable bioprocess to upcycle these rejects into polyhydroxybutyrate (PHB), a biodegradable alternative to degradation-resistant petroleum-based plastics. To achieve high yields of PHB per unit biomass, the specific objective of the study was to investigate various approaches to enhance the hydrolysis yields of fiber rejects to maximize sugar recovery and evaluate the fermentation performance of these sugars using Escherichia coli LSBJ. The investigated approaches included size reduction, surfactant addition, and a chemical-free hydrothermal pretreatment process. A two-step hydrothermal pretreatment, involving a hot water pretreatment (150 °C and 15% solid loading for 10 min) followed by three cycles of disk refining, was found to be highly effective and resulted in an 83% cellulose conversion during hydrolysis. The hydrolysate obtained from pretreated biomass normally requires a detoxification step to enhance fermentation efficiency. However, the hydrolysate obtained from the pretreated biomass contained minimal to no inhibitory compounds, as indicated by the efficient sugar fermentation and high PHB yields, which were comparable to those from fermenting raw biomass hydrolysate. The structural and thermal properties of the extracted PHB were analyzed using various techniques and consistent with standard PHB. [ABSTRACT FROM AUTHOR]

Nucleic acid amplification testing has great potential for point-of-need diagnostic testing with high detection sensitivity and specificity. Current sample preparation is limited by a tedious workflow requiring multiple steps, reagents and instrumentation, hampering nucleic acid testing at point of need. In this study, we present the use of mixed cellulose ester (MCE) paper for DNA binding by ionic interaction under molecular crowding conditions and fluid transport by wicking. The poly(ethylene) glycol-based (PEG) reagent simultaneously provides the high pH for alkaline lysis and crowding effects for ionic binding of the DNA under high salt conditions. In this study, we introduce Paper-based Abridged Solid-Phase Extraction with Alkaline Poly(ethylene) Glycol Lysis (PASAP). The anionic mixed cellulose ester (MCE) paper is used as solid phase and allows for fluid transport by wicking, eliminating the need for pipetting skills and the use of a magnet to retain beads. Following the release of DNA from the cells due to the lytic activity of the PASAP solution, the DNA binds to the anionic surface of the MCE paper, concentrating at the bottom while the sample matrix is transported towards the top by wicking. The paper was washed by dipping it in 40% isopropanol for 10 s. After air-drying for 30 s, the bottom section of the paper (3 mm × 4 mm) was snapped off using the cap of a PCR tube and immersed in the colourimetric loop-mediated isothermal amplification (cLAMP) solution for direct amplification and colourimetric detection. The total sample processing was completed in 15 min and ready for amplification. cLAMP enabled the detection of 102 CFU/mL of Escherichia coli (E. coli) from culture media and the detection of E. coli in milk < 103 CFU/mL (10 CFU) after incubation at 68 °C for 60 min, demonstrating applicability of the method to complex biological samples. [ABSTRACT FROM AUTHOR]

This research investigated the inhibition of *Escherichia coli* ATCC 25922 (E. coli) bacterial growth in situ, specifically on the stems and aerial parts of *Agastache mexicana* subsp. mexicana (Amm) or "purple toronjil" and on food-grade paper, both contained within Kraft paper bags with a plastic window. The qualitative phytochemical profile of an aqueous extract of Amm revealed the presence of various compounds including alkaloids, coumarins, tannins, flavonoids, saponins, triterpenes, and sterols. The results indicate that these secondary metabolites exhibit a synergistic bactericidal effect, especially when combined with temperature and starvation stress. This was quantified using a decay equation referred to as the bacterial growth inhibition profile of E. coli (BGIPEc). Calculations, which included first derivatives, gradients based on substrate effects and temperature as well as the area under the curve of BGIPEc, demonstrated that higher temperatures led to the greater inhibition of colony forming units (CFUs), further enhanced by the presence of secondary metabolites. Additionally, a shorter half-life corresponded to a faster change rate and a lower area under the curve, indicating a reduced survival rate over time. At lower temperatures, E. coli exhibited a survival effect, which was corroborated by the preceding calculations. [ABSTRACT FROM AUTHOR]

In the pursuit of enhancing food packaging, nanotechnology, particularly green silver nanoparticles (G-AgNPs), have gained prominence for its remarkable antimicrobial properties with high potential for food shelf-life extension. Our study aims to develop corn starch-based coating materials reinforced with G-AgNPs. The mechanical properties were examined using a uniaxial tensile tester, revealing that starch coated with the highest G-AgNPs concentration (12.75 ppm) exhibited UTS of 87.6 MPa compared to 48.48 MPa of control paper, a significant (p < 0.02) 65% increase. The assessment of the WVP showcased a statistical reduction in permeability by up to 8% with the incorporation of the hydrophobic layer. Furthermore, antibacterial properties were assessed following ISO 22196:2011, demonstrating a strong and concentration-dependent activity of G-AgNPs against E. coli. All samples successfully disintegrated in both simulated environments (soil and seawater), including samples presenting G-AgNPs. In the food trial analysis, the presence of starch and G-AgNPs significantly reduced weight loss after 6 days, with cherry tomatoes decreasing by 8.59% and green grapes by 6.77% only. The results of this study contribute to the advancement of environmentally friendly packaging materials, aligning with the UN sustainable development goals of reducing food waste and promoting sustainability. [ABSTRACT FROM AUTHOR]

The freshness and safety of fruits and vegetables affect our daily life. Paper products are often used in the packaging and transportation of fruits and vegetables, and these can provide other functions besides packaging after certain modifications and additions. In this study, the AgNPs/1-MCP antibacterial fresh-keeping composite paper was prepared by in-situ loaded silver nanoparticles and spraying 1-MCP solution. Moreover, the prepared paper was used to preserve sweet cherries. It was found that the prepared AgNPs/1-MCP antibacterial fresh-keeping composite paper could effectively inhibit E. coli and S. aureus. When the addition of 1-MCP in the paper was 0.05 g, the fresh-keeping effect on cherries was the best. Under this optimal condition, the weight loss ratio of the cherries was reduced by 1.93%, the firmness was increased by 27.7%, and the soluble solid content was increased by 25%. The preservation time was extended from 4 days to 12 days, three times that of the untreated ones. The prepared fresh-keeping material is environmentally friendly, non-toxic and harmless, simple to prepare and convenient to use, and is expected to become one of the important fresh-keeping methods for fruits. [ABSTRACT FROM AUTHOR]

Detecting alterations in plasmid structures is often performed using conventional molecular biology. However, these methods are laborious and time-consuming for studying the conditions inducing these mutations, which prevent real-time access to cell heterogeneity during bioproduction. In this work, we propose combining both flow cytometry and fluorescence-activated cell sorting, integrated with mechanistic modelling to study conditions that lead to plasmid recombination using a limonene-producing microbial system as a case study. A gene encoding GFP was introduced downstream of the key enzymes involved in limonene biosynthesis to enable real-time kinetics monitoring and the identification of cell heterogeneity according to microscopic and flow cytometric analyses. Three different plasmid configurations (one correct and two incorrect) were identified through cell sorting based on subpopulations expressing different levels of GFP at 10 and 50 µM IPTG. Higher limonene production (530 mg/L) and lower subpopulation proportion carrying the incorrect plasmid (12%) were observed for 10 µM IPTG compared to 50 µM IPTG (96 mg/L limonene and more than 70% of cell population carrying the incorrect plasmid, respectively) in 100 mL production culture. We also managed to derive exploratory hypotheses regarding the plasmid recombination region using the model and successfully validated them experimentally. Additionally, the results also showed that limonene production was proportional to GFP fluorescence intensity. This correlation could serve as an alternative to using biosensors for a high-throughput screening process. The developed method enables rapid identification of plasmid recombination at single-cell level and correlates the heterogeneity with bioproduction performance. KEY POINTS: • Strategy to study plasmid recombination during bioproduction. • Different plasmid structures can be identified and monitored by flow cytometry. • Mathematical modelling suggests specific alterations in plasmid structures., Competing Interests: Declarations. This article does not contain any studies with animals performed by any of the authors. Conflict of interest: The authors declare no competing interests., (© 2025. The Author(s).)

Chitosan, a renowned natural polymer for its wide application, was exploited for industry paper coating solutions. This research aimed to investigate the antimicrobial and water vapor barrier of chitosan solution-coated industrymanufactured paper. The papers were obtained from a national company in Indonesia. The commercially available chitosan with three molecular weight levels (low, medium, and high) was solubilized in sodium acetate buffer and subsequently utilized as the coating solution. The other variable studied was the chitosan concentration in the coating solution, i.e., 1.0, 1.5, and 2.0% (w/w). The antimicrobial activity study was performed by testing against Gram-positive bacteria, represented by Staphylococcus aureus, and Gram-negative bacteria, represented by Escherichia coli. The low molecular weight chitosan showed the best antimicrobial activity and water vapor barrier performance. The 1 %w low molecular weight chitosan-coated paper had shown good antimicrobial activity, against both S. aureus and E. coli, with a growth reduction of > 95 %. The most effective antimicrobial activity against S. aureus was achieved by paper coated with a 1.5% solution of low molecular weight chitosan. For low molecular weight chitosan-coated paper the most effective water vapor barrier was exhibited at 1 %w chitosan concentration. Having shown the best water vapor barrier while maintaining good antimicrobial activity, the 1.0% solution of low molecular weight chitosan was appointed as the best coating solution in this research. [ABSTRACT FROM AUTHOR]

Uropathogenic Escherichia coli (UPEC) causes up to 90% of urinary tract infections (UTI) which is more prevalent among females than males. In urine, patients with symptomatic UTI usually have a high concentration of bacterial infection, ≥ 105 colony-forming units (CFU) per mL, in which the culture method is regularly the gold standard diagnosis. In this study, a simple and inexpensive distance-based paper device (dPAD) combined with the fluorescent closed tube LAMP assay was validated for simultaneously screening and semi-quantifying the infection level of E. coli in 440 urine samples of patients with UTI. The dPAD could measure the LAMP amplicons and semi-quantify the levels of E. coli infection in heavy (≥ 104 CFU/mL), light (≤ 103 CFU/mL) and no infection. The sensitivity and specificity had reliable performances, achieving as high as 100 and 92.7%, respectively. The one step LAMP assay could be performed within 3 h, which was 7.5 times faster than the culture method. To empower early UTI diagnosis and fast treatment, this inexpensive dPAD tool combined with the fluorescent closed tube LAMP assay is simple, reliably fast and practically portable for point-of-care settings, particularly in resource-limited areas, which can be set up in all levels of healthcare facilities. [ABSTRACT FROM AUTHOR]

Colombia shows a high density and variety of bird species, making it one of the most diverse avian territories globally. Antioquia ranks among the top four provinces with the greatest variety of bird species, underscoring the importance of research efforts on the local bird fauna. Therefore, this study aimed to identify bacterial agents in the blood of wild birds from the municipality of Jardín (Antioquia, Colombia) by 16S qPCR sequencing. A descriptive cross-sectional study was conducted using non-probabilistic convenience sampling. Wild birds were captured using mist nets and blood samples were collected from each animal via puncture using sterile lancets in the brachial vein, and a drop of blood was collected on filter paper for qPCR analysis. The 16S gene in bacterial genomes was found in 13 out of 46 wild birds of the Passeriform and quasi-Passeriform orders, captured at three different locations within the study municipality at altitudes ranging from 1,665 to 2,034 m.a.s.l. Seven different bird species were recorded and four different haemobacteria were identified (i.e. Exiguobacterium spp., Escherichia coli, Stenotrophomonas spp., and Stenotrophomonas maltophilia). This study contributes to the knowledge in Colombia by identifying four different hemobacteria in wild birds. Further research is required on the health status of these birds and the attributable impacts on their populations and other related factors, including humans. [ABSTRACT FROM AUTHOR]

The study investigates four paper-based materials designed for short-time wrapping of meat products by determining morpho-structure, capillary-hydroscopic, barrier and antibacterial properties, wettability and migration into food simulants. The paper-based materials are coded as RO, SP, IT and SLO. RO and SLO samples exhibit the best barrier properties against water vapors. The low solubility and contact angles of RO, IT and SLO in A simulant (distilled water) make them suitable for aqueous food storage. The extremely high solubility of SP and SLO in simulant B (acetic acid) shows that wax and hydrophobized starch, respectively are carried by the acidic media, thus these agents are unlikely to coat the paper designed to package acidic food. SLO inhibits E. coli, Salmonella enterica, Lysteria monocytogenes, Pseudomonas aeruginosa and fluorescens. Polyethylene coated on RO and IT surface and wax impregnated on SP have a lower antimicrobial activity in comparison with hydrophobized starch coated on SLO. [ABSTRACT FROM AUTHOR]

Melamine-impregnated paper is made by impregnating paper with melamine-formaldehyde resin and can be hot-pressed and laminated onto the surface of wood boards. If the paper has antibacterial activity, it can prevent the spread of indoor bacteria, but impregnated paper with a long-term antibacterial effect has not been proposed. Metal–organic frameworks (MOFs) have been studied as a reservoir of metal ions. In this study, a silver-metal organic framework (Ag-BTC) was synthesized by a green and scalable method at 60 °C (A-60) and at room temperature (A-25). The structure and thermal stability of the Ag-BTC were characterized. Compared with A-25, the structure and particle size distribution of A-60 were more regular and uniform. The thermal stability and the compatibility of Ag-BTC and MF resin were suitable for hot-pressing process. Ag-BTC in the cured impregnated paper exhibited the sustained-release performance of Ag+. The antibacterial ratio of decorated blockboards with 0.10 wt% A-25 to E. coli and S. aureus exceeded 99.99%, reaching Level I antibacterial grade. Compared with A-60-decorated blockboards, the better antibacterial activity of A-25-decorated blockboards was due to the release of more Ag+. This study provides new ideas for the design of antibacterial melamine-impregnated paper. [ABSTRACT FROM AUTHOR]

Wounds often develop infections caused by microbes. Preventing these microbes from causing infection is essential for wound healing. The current study therefore focuses on the preparation of an Fp (cellulose) scaffold coated with antimicrobial clindamycin drug (Cl) and chitosan (Cs) for use in wound dressings. Fp was used as a control. Before testing the efficacy of ClCFp scaffolds against bacterial strains and evaluating their cytocompatibility, they were thoroughly characterized using microscopic, spectroscopic, BET, and X-ray techniques. These methods confirmed coating of Cs and Cl drug on Fp. No significant structural damage to the Fp was observed. The porosity and surface area were found to decrease with coating. Invitro studies with HGF cells showed that the cells not only adhered to the scaffolds and proliferated, but also exhibited good viability. The results of the antibacterial tests showed that ClCFp scaffold was effective against both E. coli, "gram-negative" bacteria, and S. aureus, "gram-positive bacteria". Docking studies performed for the transcription termination factor Rho of E. coli and penicillin-binding protein 2 of S. aureus against the Cl drug confirmed the interaction between the Cl drug and bacterial proteins. Therefore, the ClCFp composite scaffolds have good prospects for biomedical applications and especially for wound dressings. [ABSTRACT FROM AUTHOR]

[Display omitted] • A simple synthesis method has been developed for Ag@ZIF-8, involving controlling the loading and size of Ag NPs. • Ag@ZIF-8 meet the requirements for antibacterial agent used in the melamine-impregnated paper. • The multiple synergistic antibacterial mechanism of Ag@ZIF-8 was verified. • The decorated blockboards with 0.20 wt% Ag@ZIF-8 reached Level Ⅰ antibacterial grade. Melamine-impregnated paper, a decorative material in furniture and interior design, could potentially impede the spread of indoor bacteria if it possesses antibacterial properties. Here, nano Ag-doped ZIF-8 (Ag@ZIF-8) was synthesized using ion exchange and in-situ chemical reduction strategies, followed by incorporating it into melamine-impregnated paper. The ion exchange mechanism of Zn2+ and Ag+ during ion exchange reactions was investigated. The structure of Ag@ZIF-8 was well-defined, with Ag nanoparticles evenly distributed on the surface of ZIF-8. Ag@ZIF-8 showed good thermal stability at 400 °C, meeting the requirements for the hot-pressing process of the melamine-impregnated paper. Ag@ZIF-8 exhibited enhanced ROS generation under visible light compare to ZIF-8, which was attributed to its reinforced light absorption and charge carrier separation facilitated by the localized surface plasmon resonance (LSPR) effect. The antibacterial activity of Ag@ZIF-8 was better than ZIF-8, owing to the multiple-synergistic antibacterial mechanisms of the ion release and ROS generation. Specifically, the 10-Ag@ZIF-8 exhibited MIC of 128 μg/mL and 64 μg/mL against E. coli and S. aureus , respectively. The signal intensity of •O 2 − induced by 10-Ag@ZIF-8 was 4.38 times higher than that of ZIF-8. As the addition of Ag@ZIF-8 increased, the antibacterial performance of melamine-impregnated paper improved. Considering the antibacterial property and economy of melamine-impregnated paper, 0.20 wt% was the optimal amount for 10-Ag@ZIF-8 decorated blockboard. This study proposes a novel approach for synthesizing MOF-based melamine-impregnated paper with antibacterial properties, aiming to reduce bacterial spread, minimize disease risks, and contribute to lowering overall healthcare costs. [ABSTRACT FROM AUTHOR]

Microbial xylanase has wide application in bioenergy, animal feed, environmental protection, the pulp and paper industry, and agricultural development. In this study, three xylanases from the microbiota of T. fuciformis with its companion strains were identified by metagenomics sequencing. The three enzymes were subjected to cloning and expression in E. coli or P. pastoris, purification, and characterization for their properties. The results showed that AsXyn1, from Annulohypoxylon stygium, among the three enzymes possessed high thermostability at 40 °C and broad pH tolerance in the range of 2.0–10.0, exhibiting its application potential. Furthermore, it was found that post-translational modification (such as glycosylation) of AsXyn1 enzyme modulated its activity, kinetic parameters, and thermostability. These results and findings provided a hint for enzyme modification and design in future. [ABSTRACT FROM AUTHOR]

Insufficiently treated wastes contained within onsite sanitation systems (OSS) commonly used by disadvantaged and developing communities contribute to public and environmental health concerns, calling for practical alternative solutions. At the basic level, an improved understanding of the evolution of chemical and physical constituents under different waste introduction methods and for short-term and long-term operation is needed. While receiving non-dilute waste under mixed, unmixed, toilet paper exclusion, and urine diversion (UD) regimes, self-flushing OSS simulated using anaerobic digesters (ADs) were compared during three operational stages: (1) 0–1 month service for unsheltered encampments; (2) 1–3 month disaster relief scenario; and (3) ≥ 3 months representing refugee camps and long-term household use. Although stratification was found to promote suitable conditions for short-term use of self-flushing toilets, mixing increased beneficial biodegradation of organic constituents. Urine-containing ADs demonstrated a shift from sulfide to ammonia odor accompanied by high pH (> 8) after ~ 240 d. E. coli reduction following elevated nitrogen and dissolved solids levels pointed to decreased pathogen survival in ADs with urine. The benefits of bacterial disinfection, reduction of sulfurous odors, and heightened organics degradation in mixed, urine-containing ADs suggest this format as more desirable for prolonged use of self-flushing OSS over unmixed or urine-diverting formats. [ABSTRACT FROM AUTHOR]

The rapid and specific point-of-care (POC) analysis of virulent pathogenic strains plays a key role in ensuring food quality and safety. In this work, a paper-based fluorescent phage biosensor was developed for the detection of the virulent E. coli O157:H7 strain (as the mode of virulent pathogens) in food samples. Firstly, phages that can specifically combine with E. coli O157:H7 (E. coli) were stained with SYTO-13 dye to prepare a novel fluorescent probe (phage@SYTO). Simultaneously, a micro-porous membrane filter with a pore size of 0.45 μm was employed as a paper chip so as to retain the E. coli-phage@SYTO complex (>1.2 μm) on its surface. The phage@SYTO (200 nm in size) was able to pass through the pores of the chip, and the complex could be retained on the paper chip using the free phage@SYTO probes. The E. coli-phage@SYTO could emit a visual fluorescent signal (excited at 365 nm; emitted at 520 nm) onto the chip, which could be detected by a smartphone to reflect the concentration of E. coli. Under optimized conditions, the detection limit was as low as 50 CFU/mL (S/N = 3) and exhibited a wide linear range from 102 to 106 CFU/mL. The sensor has potential application value for the quick and specific POCT detection of virulent E. coli in foods. [ABSTRACT FROM AUTHOR]

Global food systems can benefit significantly from continuous monitoring of microbial food safety, a task for which tedious operations, destructive sampling, and the inability to monitor multiple pathogens remain challenging. This study reports significant improvements to a paper chromogenic array sensor - machine learning (PCA-ML) methodology sensing concentrations of volatile organic compounds (VOCs) emitted on a species-specific basis by pathogens by streamlining dye selection, sensor fabrication, database construction, and machine learning and validation. This approach enables noncontact, time-dependent, simultaneous monitoring of multiple pathogens (Listeria monocytogenes , Salmonella , and E. coli O157:H7) at levels as low as 1 log CFU/g with over 90% accuracy. The report provides theoretical and practical frameworks demonstrating that chromogenic response, including limits of detection, depends on time integrals of VOC concentrations. The paper also discusses the potential for implementing PCA-ML in the food supply chain for different food matrices and pathogens, with species- and strain-specific identification. [ABSTRACT FROM AUTHOR]

Background: The utilization of antibiotic adjuvants presents a promising strategy for addressing bacterial resistance. Recently, the development of antibiotic adjuvants has attracted considerable attention from researchers in academia and industry. This study aimed to identify the most influential publications on antibiotic adjuvants and elucidate the hotspots and research trends in this field. Method: Original articles and reviews related to antibiotic adjuvants were retrieved from the Web of Science Core Collection database. The top 100 highly cited publications were selected and the visual analyses of publication outputs, countries, institutions, authors, journals, and keywords were conducted using Excel, VOSviewer, or CtieSpace software tools. Results: The top 100 cited publications concerning antibiotic adjuvants spanned the years 1977-2020, with citation counts ranging from 174 to 2,735. These publications encompassed 49 original articles and 51 reviews. The journal "Antimicrobial Agents and Chemotherapy" accounted for the highest number of publications (12%). The top 100 cited publications emanated from 39 countries, with the United States leading in production. Institutions in Canada and the United States exhibited the most substantial contributions to these highly cited publications. A total of 526 authors participated in these studies, with Robert E. W. Hancock, Laura J. V. Piddock, Xian-Zhi Li, Hiroshi Nikaido, and Olga Lomovskaya emerging as the most frequently nominated authors. The most common keywords included "E. coli", "P. aeruginosa", "S. aureus", "in-vitro activity", "antimicrobial peptide", "efflux pump inhibitor" "efflux pump", "MexAB-OprM" and "mechanism". These keywords underscored the hotspots of bacterial resistance mechanisms and the development of novel antibiotic adjuvants. Conclusion: Through the bibliometric analysis, this study identified the top 100 highly cited publications on antibiotic adjuvants. Moreover, the findings offered a comprehensive understanding of the characteristics and frontiers in this field. [ABSTRACT FROM AUTHOR]

Biological toxicity testing plays an essential role in identifying the possible negative effects induced by substances such as organic pollutants or heavy metals. As an alternative to conventional methods of toxicity detection, paper-based analytical device (PAD) offers advantages in terms of convenience, quick results, environmental friendliness, and cost-effectiveness. However, detecting the toxicity of both organic pollutants and heavy metals is challenging for a PAD. Here, we show the evaluation of biotoxicity testing for chlorophenols (pentachlorophenol, 2,4-dichlorophenol, and 4-chlorophenol) and heavy metals (Cu2+, Zn2+, and Pb2+) by a resazurin-integrated PAD. The results were achieved by observing the colourimetric response of bacteria (Enterococcus faecalis and Escherichia coli) to resazurin reduction on the PAD. The toxicity responses of E. faecalis-PAD and E. coli-PAD to chlorophenols and heavy metals can be read within 10 min and 40 min, respectively. Compared to the traditional growth inhibition experiments for toxicity measuring which takes at least 3 h, the resazurin-integrated PAD can recognize toxicity differences between studied chlorophenols and between studied heavy metals within 40 min. [ABSTRACT FROM AUTHOR]

Summary: LAMP assay is widely used for detecting pathogens. We observed that the conventional and gradient polymerase chain reaction (PCR) could not detect the extracted Escherichia coli DNA; real-time PCR was able to detect up to a certain limit (10-8 bacterial dilution). At the same time, the LAMP assay could detect the bacteria at a much lower concentration (10-14 dilution). The results of the LAMP assay were evaluated using agarose gel electrophoresis and DNA binding dye (PicoGreen), but only gel electrophoresis gave reliable results. Therefore, we propose using electrophoresis-based amplicon detection to overcome the limitations of dye-based detection. We believe that this amplicon detection will go a long way in the screening of potable drinking water., (Copyright © 2023 Copyright: © 2023 Indian Journal of Public Health.)

In microbial cell factory, CO 2 release during acetyl-CoA production from pyruvate significantly decreases the carbon atom economy. Here, we construct and optimize a synthetic carbon conserving pathway named as Sedoheptulose-1,7-bisphosphatase Cycle with Trifunctional PhosphoKetolase (SCTPK) in Escherichia coli. This cycle relies on a generalist phosphoketolase Xfspk and converts glucose into the stoichiometric amounts of acetylphosphate (AcP). Furthermore, genetic circuits responding to AcP positively or negatively are created. Together with SCTPK, they constitute a gene-metabolic oscillator that regulates Xfspk and enzymes converting AcP into valuable chemicals in response to intracellular AcP level autonomously, allocating metabolic flux rationally and improving the carbon atom economy of bioconversion process. Using this synthetic machinery, mevalonate is produced with a yield higher than its native theoretical yield, and the highest titer and yield of 3-hydroxypropionate via malonyl-CoA pathway are achieved. This study provides a strategy for improving the carbon yield of microbial cell factories., (© 2023. Springer Nature Limited.)

The outer membrane structure is common in Gram-negative bacteria, mitochondria and chloroplasts, and contains outer membrane β-barrel proteins (OMPs) that are essential interchange portals of materials 1-3 . All known OMPs share the antiparallel β-strand topology 4 , implicating a common evolutionary origin and conserved folding mechanism. Models have been proposed for bacterial β-barrel assembly machinery (BAM) to initiate OMP folding 5,6 ; however, mechanisms by which BAM proceeds to complete OMP assembly remain unclear. Here we report intermediate structures of BAM assembling an OMP substrate, EspP, demonstrating sequential conformational dynamics of BAM during the late stages of OMP assembly, which is further supported by molecular dynamics simulations. Mutagenic in vitro and in vivo assembly assays reveal functional residues of BamA and EspP for barrel hybridization, closure and release. Our work provides novel insights into the common mechanism of OMP assembly., (© 2023. The Author(s), under exclusive licence to Springer Nature Limited.)

Shellfish-growing areas in rural catchments are occasionally affected by elevated faecal contamination from diffuse sources and may be subject to frequent harvest closures/classification downgrades. We combined traditional risk management methods based on sanitary surveys and monitoring of Escherichia coli in seawater and shellfish with faecal source tracking, bacterial source apportionment, and hydrometeorological modelling to determine the causes of elevated E. coli concentrations contributing to harvest closures in Papanui Inlet (Aotearoa New Zealand). These multiple lines of evidence were used to inform a weight of evidence assessment of bacterial contamination in the inlet. Ruminant livestock was estimated to contribute 80% of the faecal coliform loading. Concentrations of E. coli in seawater were low (≤ 87 MPN 100 ml -1 ) whilst concentrations in tuaki/cockles/little neck clams (Austrovenus stutchburyi) occasionally exceeded the "Approved" classification limit (230 MPN 100 g -1 ). The most frequent positive genetic markers in seawater were the seagull (Catellicoccus marimammalium) (54% of seawater samples), and in shellfish, the bovine and seagull markers (both 12.5% of shellfish samples). Solar radiation was negatively correlated with E. coli in tuaki. We found that the growing area is affected by faecal inputs from animal and, to a lesser extent, human (septic tank discharges) sources which elevate contamination to levels detectable in tuaki but not in seawater, particularly in the summer months. The innovative approach can enhance the management of shellfish-growing areas affected by intermittent contamination and enables more targeted action to reduce pollution to improve shellfish water quality., (© 2023. The Author(s), under exclusive licence to Springer Nature Switzerland AG.)

Total coliforms, E. coli, and fecal streptococci are the important indicators linked to the human health. This study investigated presence of these indicator bacteria in the Himalayan springs at various locations in the district Kulgam of Kashmir valley. A total of 30 spring water samples were collected from rural, urban, and forest areas during post-melting season 2021 and pre-melting season 2022. The springs in the area originate from the alluvium deposit, Karewa, and hard rock formations. The physicochemical parameters were found within the acceptable limits. However, nitrate and phosphate were found above the permissible limit at few sites, thus indicating the presence of anthropogenic activities in the area. Majority of the samples during both the seasons were found highly loaded with total coliforms with maximum limit of greater than 180 MPN/100 ml. E. coli and fecal streptococci were found in the range of less than 1 to more than 180 MPN/100 ml. The results of Pearson correlation of physicochemical parameters with indicator bacteria showed that chemical oxygen demand, rainfall, spring discharge, nitrate, and phosphate are the main factors affecting the concentration of indicator bacteria in the spring water at each site. Principal component analysis showed the most influencing factors of water quality in most of the spring sites are total coliforms, E. coli, fecal streptococci, rainfall, discharge, and chemical oxygen demand. The results of this study showed that the spring water is unfit for drinking purpose because of high concentration of fecal indicator bacteria., (© 2023. The Author(s), under exclusive licence to Springer Nature Switzerland AG.)

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Chitosan/lignin nanocomposites were coated on bagasse paper surface to improve antimicrobial packaging. A commercial softwood kraft lignin was selected and purified by fractionation process using organic solvent. The aliphatic, phenolic and carboxylic hydroxyl groups of commercial softwood lignin and fractionated lignin were investigated using 31P NMR spectroscopy. For lignin nanosphere particles (LNPs) preparation, the fractionated lignin was dissolved in organic solvent (acetone), then the deionized water was added under ultrasonic mixing. The size of LNPs were approximately 40–500 nm. Chitosan solutions were mixed with 6–33 wt% of LNPs. The mixture was then spread over the bagasse paper surface. The papers coated with chitosan/LNPs were dehydrated at 40 °C for 48 h. For food packaging applications, the antibacterial activity of the paper packaging coated with chitosan/LNPs was more successful and effective against gram-positive bacteria (such as Staphylococcus aureus, S. aureus) and gram-negative bacteria (such as Escherichia coli, E. coli) compared with the bagasse paper without any coating. The antimicrobial activity of all papers coated with chitosan/LNPs for S. aureus and E. coli was above 2 log CFU/cm2 which are acceptable for antimicrobial efficacy and the percent reduction of bacteria was nearly 100%. In addition, this evident of chitosan/LNPs nanocomposites showed a synergistic effect enhancing the antimicrobial efficiency on the paper surface. The morphology, wettability and overall migration of paper coated with chitosan/LNPs samples were also explored. [Display omitted] [ABSTRACT FROM AUTHOR]

Organophosphates are becoming an emerging pollutant due to their various applications, particularly as pesticides. In this study, an improved Colony (Live-cell) PCR method was developed for the detection of opd genes from bacteria encoding the organophosphate hydrolase enzymes capable of degrading various organophosphates. The improved method does not require pre-heating or pre-lysis of bacterial cells as essential in the conventional colony PCR. The reaction volume was scaled down to 10 µl by optimizing the PCR buffer and amplification conditions. The improved method was used for Gram positive and negative bacteria, glycerol stocks, liquid cultures, recombinant and mutant strains. Also, 16S rRNA gene was amplified from unknown environmental isolates and known E. coli strains. The amplified opd and 16S rRNA genes from the improved colony PCR method and by conventional PCR were sequenced, and similar results were obtained from both techniques. Thus, the improved method can be further explored in molecular biology or during biomarker studies. KEY POINTS: • Improved colony PCR method was developed for screening of opd genes from bacteria. • Method was validated for Gram positive/negative bacteria from solid as well as liquid media. • The improved method was rapid, efficient, and can be applied under various conditions., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Foodborne pathogens are major public health concerns worldwide. Paper-based microfluidic devices are versatile, user friendly and low cost. We report a novel paper-based single input channel microfluidic device that can detect more than one whole-cell foodborne bacteria at the same time, and comes with quantitative reading via image analysis. This microfluidic paper-based multiplexed aptasensor simultaneously detects E. coli O157:H7 and S. Typhimurium. Custom designed particles provide colorimetric signal enhancement and false results prevention. Several aptamers were screened and the highest-affinity aptamers were optimized and employed for detection of these bacteria in solution, both in a buffer as well as pear juice. Image analysis was used to read and quantify the colorimetric signal and measure bacteria concentration, thus rendering this paper based microfluidic device quantitative. The colorimetric results show linearity over a wide concentration range (102 C F U / m L to 108 C F U / m L) and a limit of detection (LOD) of 103 C F U / m L and 102 C F U / m L for E. coli O157:H7 and S. Typhimurium, respectively. An insignificant change in colorimetric response for non-target bacteria indicates the aptasesnors are specific. The reported multiplexed colorimetric paper-based microfluidic devices is likely to perform well for on-site rapid screening of pathogenic bacteria in water and food products. [ABSTRACT FROM AUTHOR]

Microbial infections are a global health challenge often treated with antibiotics, but their overuse can lead to antibiotic resistance. To address this, there is a need for more efficient personal protective equipment (PPE) with novel antimicrobial techniques. One such technique involves using carbon dots (CDs) to create self-disinfecting fabrics. This study focused on synthesizing phenylboronic-acid-functionalized piperine-based carbon dots (PBA-PPCDs) and boronic acid carbon dots (BACDs) as potential antimicrobial agents. The materials were successfully synthesized using a hydrothermal method and characterized using various techniques to confirm the size, charge, fluorescence properties, and inner morphology of the CDs. The CDs showed bactericidal activity against gram-positive and gram-negative bacteria, with potent inhibition in growth especially in the presence of simulated sunlight. The antibacterial properties of the CDs were tested on cellulose discs, demonstrating strong adherence and significant inhibition of bacterial cells. The cytotoxicity studies showed that these CDs, at concentrations up to 0.1 mg/mL, and the CD-modified fibers are benign to mammalian cells, emphasizing their safety for human use. Overall, the study concluded that PBA-PPCDs and BACDs are promising candidates for modifying fibers and inducing antimicrobial activity in fabrics made of cellulose fibers.Article highlights: New self-disinfecting fabrics kill bacteria with light-activated carbon dots, no harsh chemicals needed. Self-disinfecting fabrics fight bacteria, reduce antibiotic use, and slow superbug growth. Safe and effective: Carbon dot fabrics harmless to humans, promising for PPE. [ABSTRACT FROM AUTHOR]

Escherichia coli (E. coli) cells are present in fecal materials that can be the main source for disease‐causing agents in water. As a result, E. coli is recommended as a water quality indicator. We have developed an innovative platform to detect E. coli for monitoring water quality on-site by integrating paper-based sample preparation with nucleic acid isothermal amplification. The platform carries out bacterial lysis and DNA enrichment onto a paper pad through ball-based valves for fluid control, with no need of laboratory equipment, followed by loop-mediated isothermal amplification (LAMP) in a battery-operated coffee mug, and colorimetric detection. We have used the platform to detect E. coli in environmental water samples in about 1 h, with a limit of quantitation of 0.2 CFU/mL, and 3 copies per reaction. The platform was confirmed for detecting multiple E. coli strains, and for water samples of different salt concentrations. We validated the functions of the platform by analyzing recreational water samples collected near the Atlantic Ocean that contain different concentrations of salt and bacteria. [ABSTRACT FROM AUTHOR]

The continuous emergence of new illegal compounds, particularly psychoactive chemicals, poses significant challenges for current drug detection methods. Developing new protocols and kits for each new drug requires substantial time, effort, and dedicated manpower. Whole-cell bacterial bioreporters have been proven capable of detecting diverse hazardous compounds in both laboratory and field settings, identifying not only single compounds but also chemical families. We present the development of a microbial bioreporter for the detection of cocaine, the nervous system stimulant that is the second-most widely used illegal drug in the US. Escherichia coli was transformed with a plasmid containing a bacterial luxCDABEG bioluminescence gene cassette, activated by a cocaine-responsive signaling cascade. The engineered bioreporter is demonstrated to be a sensitive and specific first-generation detection system for cocaine, with detection thresholds of 17 ± 8 μg/L and 130 ± 50 μg/L in a buffer solution and in urine, respectively. Further improvement of the sensor's performance was achieved by altering the nucleotide sequence of the PBen gene promoter, the construct's sensing element, using accelerated site-directed evolution. The applicability of ready-to-use paper strips with immobilized bioreporter cells was demonstrated for cocaine detection in aqueous solutions. [ABSTRACT FROM AUTHOR]

Hybrid kinetic models, linking structured models of cell (nano-scale) metabolic processes to the dynamics of macroscopic variables of the bioreactor, are proven to lead to more precise predictions of all key-species dynamics under variable operating conditions, being of an exceptional importance in engineering evaluations to in-silico (math-model-based) determine the optimal operating mode of a fed-batch bioreactor (FBR). Even if such extended dynamic models require more experimental and computational efforts, their use has proven to be advantageous. The approached probative example refers to the simulation of the dynamics of some key species of the central carbon metabolism (CCM) of a modified E. coli cell, linked to the state variables of a FBR used for the tryptophan (TRP) production. By using several optimization algorithms, and an original application of the Pareto-optimal front technique, this paper compares various operating alternatives by using multiple control variables, aiming to maximize TRP production, with minimum substrate consumption. The used E. coli strain was modified to drastically amplify the glucose (GLC) uptake into the cell. [ABSTRACT FROM AUTHOR]

Background: Understanding the aetiological organisms causing maternal infections is crucial to inform antibiotic treatment guidelines, but such data are scarce from Sub-Saharan Africa (SSA). We performed this systematic review and meta-analysis to address this gap. Methods: Microbiologically confirmed maternal infection data were collected from PubMed, Embase, and African Journals online databases. The search strategy combined terms related to bacterial infection, pregnancy, postnatal period, observational studies, SSA. Exclusion criteria included colonization, asymptomatic infection, and screening studies. Pooled proportions for bacterial isolates and antimicrobial resistance (AMR) were calculated. Quality and completeness of reporting were assessed using the Newcastle–Ottawa and STROBE checklists. Findings: We included 14 papers comprising data from 2,575 women from four sources (blood, urine, surgical wound and endocervical). Mixed-growth was commonly reported at 17% (95% CI: 12%-23%), E. coli from 11%(CI:10%-12%), S. aureus from 5%(CI: 5%-6%), Klebsiella spp. at 5%(CI: 4%- 5%) and Streptococcus spp. at 2%(CI: 1%-2%). We observed intra-sample and inter-sample heterogeneity between 88–92% in all meta-analyses. AMR rates were between 19% -77%, the highest with first-line beta-lactam antibiotics. Convenience sampling, and limited reporting of laboratory techniques were areas of concern. Interpretation: We provide a comprehensive summary of microbial aetiology of maternal infections in SSA and demonstrate the paucity of data available for this region. We flag the need to review the current local and international empirical treatment guidelines for maternal bacterial infections in SSA because there is high prevalence of AMR among common causative bacteria. Funding: This research was supported by the NIHR-Professorship/NIHR300808 and the Wellcome-Strategic-award /206545/Z/17/Z. Trial registration: Prospero ID CRD42021238515. [ABSTRACT FROM AUTHOR]

Background: The emergence and rapid spread of gram-negative bacteria resistant to carbapenems among newborns is concerning on a global scale. Nonetheless, the pooled estimate of gram-negative bacteria resistant to carbapenem that cause neonatal sepsis in developing nations remains unknown. Thus, this study aimed to determine the combined prevalence of gram-negative bacteria resistant to carbapenem in African newborns who were suspected of having sepsis. Methods: All studies published from January 1, 2010, up to December 30, 2023, from PubMed, Science Direct, Scopus electronic databases, and the Google Scholar search engine were researched. Isolates tested for carbapenem from neonates with sepsis, English language papers conducted in Africa, and cross-sectional and cohort studies papers were included. Using PRISMA guidelines, we systematically reviewed and meta-analyzed studies that assessed the prevalence of carbapenem-resistant gram-negative bacteria. The "Joanna Briggs Institute" was used critically to evaluate the quality of the included studies. The data analysis was carried out using STATA™ version 17. Heterogeneity across the studies was evaluated using Q and I 2 tests. The subgroup analysis was done and, funnel plot and Egger's regression test were used to detect publication bias. A sensitivity analysis was conducted. Results: All 36 studies were included in the meta-analysis and systematic review. The pooled prevalence of carbapenem resistance in Africa was 30.34% (95% CI 22.03–38.64%). The pooled estimate of gram-negative bacteria resistant to imipenem, and meropenem was 35.57% (95% CI 0.67–70.54%) and 34.35% (95% CI 20.04% – 48.67%), respectively. A. baumannii and Pseudomonas spp. had pooled prevalence of 45.9% (95% CI 33.1–58.7%) and 43.0% (95% CI 23.0–62.4%), respectively. Similarly, Pseudomonas spp. and A. baumannii also exhibited strong meropenem resistance, with a pooled prevalence of 29.2% (95% CI 4.8–53.5%) and 36.7% (95% CI 20.1–53.3%), respectively. E. coli and K. pneumoniae were the two most common isolates. Conclusion: There should be urgent antimicrobial stewardship practices, strengthened surveillance systems and effective treatment for neonates with sepsis. There was remarkable variation in resistance across the continent. [ABSTRACT FROM AUTHOR]

(1) Background: The complexity, amount of time, and the large amount of resource required to perform gold-standard bacteria culture procedures makes it difficult to perform timely pathogenic analyses, especially in areas where such resources are not readily available. A paper-based biochemical analytical tool can potentially tackle problems economically in terms of time and convenience, potentially finding utility in applications where simple and timely detection of bacteria is necessary; (2) Methods: The utility of paper-based MTT-PMS strips was tested using a simple colorimetric analytical methodology; (3) Results: Sufficient evidence was obtained to suggest that the strips can potentially be used as a rapid and convenient early, alternative bacteria screening tool for a variety of applications; (4) Conclusions: The potential of strips for the rapid detection of bacteria compared to standard bacteria culture is a key advantage in certain clinical, agricultural, and environmental applications. [ABSTRACT FROM AUTHOR]

Chronic wounds cause serious health and economic burdens on patients and society. Herein, a wireless and flexible smart wound dressing was developed for matrix metalloproteinase-9 (MMP-9) monitoring and antimicrobial treatment toward chronic wound management. The highly sensitive radio frequency MMP-9 sensor was realized based on a bioresponsive hydrogel with the bioactive peptide sequences. Taking advantage of the flexible inductive-capacitive (LC) circuit and bioresponsive hydrogel, the wireless and wearable smart wound dressing offered an efficient strategy for in-situ wound analysis. Besides, the controlled release of silver nanoparticles (AgNPs) from the degradable hydrogel exhibited significant antimicrobial efficacy against typical bacteria in wound infection including Escherichia coli and Staphylococcus aureus. The analysis of MMP-9 in wound exudate from diabetic foot ulcer (DFU) patients demonstrated good accuracy cross-validated with gold-standard fluorescent measurements, providing great potential for personalized wound management., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Phototherapy has emerged as a potential treatment strategy for bacteria-infected wounds, but the inadequate bacteria-capturing ability and excessive damage to normal tissues from single phototherapy are huge limitations. To solve the issues, herein we report the design of chitosan-based hydrogel with bacteria capturing and combined photothermal/photodynamic sterilization functions. Such hydrogel is prepared by mixing chitosan (CS) as matrix, protoporphyrin (PpIX) as photosensitizer and polydopamine (PDA) as photothermal agent and then chemically cross-linking CS with glutaraldehyde. The resulting CS-PpIX-PDA hydrogel possesses a porous architecture (average pore porosity = 60.9 %), excellent swelling capabilities (swelling ratio = 1855 %) and rheological property (G' > G″). The hydrogel can effectively produce reactive oxygen species (ROS) under 660 nm light irradiation due to the photodynamic effect of PpIX. Owing to the presence of PDA, the hydrogel displays a photoabsorption range between 600 and 1500 nm and can generate maximal temperature of 60 °C within 10 min under 808 nm laser illumination (0.6 W/cm 2 ) through photothermal effect. Besides, under synergetic illumination of 808/660 nm laser, CS-PpIX-PDA hydrogel can induce the death of 99.9999 % of E. coli and 99.99999 % of S. aureus. Importantly, when coated on the wound site, the hydrogel exhibits a remarkable bacteria-trapping ability due to its porous structure and the presence of amino groups on chitosan. Under the excitation of 660/808 nm, the combined photothermal and photodynamic effects can effectively eradicate bacteria. Simultaneously, the hydrogel also demonstrates anti-inflammatory properties and upregulates Heat Shock Protein 90 (HSP90) expression, thereby promoting collagen deposition and facilitating wound healing. Therefore, the study may provide some new insights into the development of multifunctional hydrogel for photothermal-oxidation sterilization of bacteria-infected wound therapy., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Inc. All rights reserved.)

This study was undertaken to develop a high-throughput screening strategy using a whole-cell biosensor to enhance methyl-group transfer, a rate-limiting step influenced by intracellular methyl donor availability and methyltransferase efficiency. An l-homocysteine biosensor was designed based on regulatory protein MetR from Escherichia coli, which rapidly reported intracellular l-homocysteine accumulation resulted from S-adenosyl-l-homocysteine (SAH) formation after methyl-group transfer. Using S-adenosyl-l-methionine (SAM) as a methyl donor, this biosensor was applied to caffeic acid 3-O-methyltransferase derived from Arabidopsis thaliana (AtComT). After several rounds of directed evolution, the modified enzyme achieved a 13.8-fold improvement when converting caffeic acid to ferulic acid. The best mutant exhibited a 5.4-fold improvement in catalytic efficiency. Characterization of beneficial mutants showed that improved O-methyltransferase dimerization greatly contributed to enzyme activity. This finding was verified when we switched and compared the N-termini involved in dimerization across different sources. Finally, with tyrosine as a substrate, the evolved AtComT mutant greatly improved ferulic acid biosynthesis, yielding 3448 mg L -1 with a conversion rate of 88.8%. These results have important implications for high-efficiency O-methyltransferase design, which will greatly benefit the biosynthesis of a wide range of natural products. In addition, the l-homocysteine biosensor has the potential for widespread applications in evaluating the efficiency of SAM-based methyl transfer., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)