Enhanced removal of pesticide micropollutant and bacteria using solar light-assisted Ag-doped TiO2: prospects for environmental and health impacts.

Pesticide micropollutants like 4-chlorophenol (4CP) and E. coli bacteria represent a substantial hazard, impacting both the environment and human health. This study delves into the effectiveness of Ag-doped TiO2 (Ag@TiO2) in removing both 4CP and E. coli. Ag@TiO2 has demonstrated remarkable effectiveness in removing 4CP under both solar and visible light conditions, earning degradation efficiencies of 91.3% and 72.8%, respectively. Additionally, it demonstrates outstanding photodegradation efficiency for 4CP (98.8%) at an initial concentration of 1 mg L-1. Moreover, Ag@TiO2 exhibited substantially higher removal performance for 4CP (81.6%) compared to TiO2 (27.6%) in wastewater. Analysis of the radicals present during the photodegradation process revealed that ·O2- primarily drives the decomposition of 4CP, with h+ and ·OH also playing significant roles in the oxidation reactions of the pollutant. Interestingly, even under dark conditions, Ag@TiO2 exhibited the capability to eliminate approximately 20% of E. coli, a percentage that increased to over 96% under solar light. In addition, the prospects for environmental and health impacts of utilizing Ag@TiO2 for pesticide micropollutant removal and bacteria were discussed.

Efficient photocatalytic bactericidal performance of green-synthesised TiO2/reduced graphene oxide using banana peel extracts.

In this study, the fabrication of titanium dioxide/reduced graphene oxide (TiO2/rGO) utilising banana peel extracts (Musa paradisiaca L.) as a reducing agent for the photoinactivation of Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus) was explored. The GO synthesis was conducted using a modified Tour method, whereas the production of rGO involved banana peel extracts through a reflux method. The integration of TiO2 into rGO was achieved via a hydrothermal process. The successful synthesis of TiO2/rGO was verified through various analytical techniques, including X-ray diffraction (XRD), gas sorption analysis (GSA), Fourier-transform infrared (FT-IR) spectroscopy, ultraviolet-visible diffuse reflectance spectroscopy (UV-Vis DRS), scanning electron microscope-energy dispersive X-ray (SEM-EDX) and transmission electron microscopy (TEM) analyses. The results indicated that the hydrothermal-assisted green synthesis effectively produced TiO2/rGO with a particle size of 60.5 nm. Compared with pure TiO2, TiO2/rGO demonstrated a reduced crystallite size (88.505 nm) and an enhanced surface area (22.664 m2/g). Moreover, TiO2/rGO featured a low direct bandgap energy (3.052 eV), leading to elevated electrical conductivity and superior photoconductivity. To evaluate the biological efficacy of TiO2/rGO, photoinactivation experiments targeting E. coli and S. aureus were conducted using the disc method. Sunlight irradiation emerged as the most effective catalyst, achieving optimal inactivation results within 6 and 4 h.

Experimental and Theoretical Brownian Dynamics Analysis of Ion Transport During Cellular Electroporation of E. coli Bacteria.

Escherichia coli bacterium is a rod-shaped organism composed of a complex double membrane structure. Knowledge of electric field driven ion transport through both membranes and the evolution of their induced permeabilization has important applications in biomedical engineering, delivery of genes and antibacterial agents. However, few studies have been conducted on Gram-negative bacteria in this regard considering the contribution of all ion types. To address this gap in knowledge, we have developed a deterministic and stochastic Brownian dynamics model to simulate in 3D space the motion of ions through pores formed in the plasma membranes of E. coli cells during electroporation. The diffusion coefficient, mobility, and translation time of Ca2+, Mg2+, Na+, K+, and Cl- ions within the pore region are estimated from the numerical model. Calculations of pore's conductance have been validated with experiments conducted at Gustave Roussy. From the simulations, it was found that the main driving force of ionic uptake during the pulse is the one due to the externally applied electric field. The results from this work provide a better understanding of ion transport during electroporation, aiding in the design of electrical pulses for maximizing ion throughput, primarily for application in cancer treatment.

Corrigendum: The main protease 3CLpro of the SARS-CoV-2 virus: how to turn an enemy into a helper.

[This corrects the article DOI: 10.3389/fbioe.2023.1187761.].

The main protease 3CLpro of the SARS-CoV-2 virus: how to turn an enemy into a helper.

Despite the long history of use and the knowledge of the genetics and biochemistry of E. coli, problems are still possible in obtaining a soluble form of recombinant proteins in this system. Although, soluble protein can be obtained both in the cytoplasm and in the periplasm of the bacterial cell. The latter is a priority strategy for obtaining soluble proteins. The fusion protein technology followed by detachment of the fusion protein with proteases is used to transfer the target protein into the periplasmic space of E. coli. We have continued for the first time to use the main viral protease 3CL of the SARS-CoV-2 virus for this purpose. We obtained a recombinant 3CL protease and studied its complex catalytic properties. The authenticity of the resulting recombinant enzyme, were confirmed by specific activity analysis and activity suppression by the known low-molecular-weight inhibitors. The catalytic efficiency of 3CL (0.17 ± 0.02 µM-1-s-1) was shown to be one order of magnitude higher than that of the widely used tobacco etch virus protease (0.013 ± 0.003 µM-1-s-1). The application of the 3CL gene in genetically engineered constructs provided efficient specific proteolysis of fusion proteins, which we demonstrated using the receptor-binding domain of SARS-CoV-2 spike protein and GST fusion protein. The solubility and immunochemical properties of RBD were preserved. It is very important that in work we have shown that 3CL protease works effectively directly in E. coli cells when co-expressed with the target fusion protein, as well as when expressed as part of a chimeric protein containing the target protein, fusion partner, and 3CL itself. The results obtained in the work allow expanding the repertoire of specific proteases for researchers and biotechnologists.

Antimicrobial and mechanical properties of functionalized textile by nanoarchitectured photoinduced Ag@polymer coating.

The control of microbial proliferation is a constant battle, especially in the medical field where surfaces, equipment, and textiles need to be cleaned on a daily basis. Silver nanoparticles (AgNPs) possess well-documented antimicrobial properties and by combining them with a physical matrix, they can be applied to various surfaces to limit microbial contamination. With this in mind, a rapid and easy way to implement a photoinduced approach was investigated for textile functionalization with a silver@polymer self-assembled nanocomposite. By exposing the photosensitive formulation containing a silver precursor, a photoinitiator, and acrylic monomers to a UV source, highly reflective metallic coatings were obtained directly on the textile support. After assessing their optical and mechanical properties, the antimicrobial properties of the functionalized textiles were tested against Escherichia coli (E. coli) and Candida albicans (C. albicans) strains. In addition to being flexible and adherent to the textile substrates, the nanocomposites exhibited remarkable microbial growth inhibitory effects.

Atmospheric Pressure Plasma Polymerisation of D-Limonene and Its Antimicrobial Activity.

Antibacterial coating is necessary to prevent biofilm-forming bacteria from colonising medical tools causing infection and sepsis in patients. The recent coating strategies such as immobilisation of antimicrobial materials and low-pressure plasma polymerisation may require multiple processing steps involving a high-vacuum system and time-consuming process. Some of those have limited efficacy and durability. Here, we report a rapid and one-step atmospheric pressure plasma polymerisation (APPP) of D-limonene to produce nano-thin films with hydrophobic-like properties for antibacterial applications. The influence of plasma polymerisation time on the thickness, surface characteristic, and chemical composition of the plasma-polymerised films was systematically investigated. Results showed that the nano-thin films deposited at 1 min on glass substrate are optically transparent and homogenous, with a thickness of 44.3 ± 4.8 nm, a smooth surface with an average roughness of 0.23 ± 0.02 nm. For its antimicrobial activity, the biofilm assay evaluation revealed a significant 94% decrease in the number of Escherichia coli (E. coli) compared to the control sample. More importantly, the resultant nano-thin films exhibited a potent bactericidal effect that can distort and rupture the membrane of the treated bacteria. These findings provide important insights into the development of bacteria-resistant and biocompatible coatings on the arbitrary substrate in a straightforward and cost-effective route at atmospheric pressure.

NiFe2O4/g-C3N4 heterostructure with an enhanced ability for photocatalytic degradation of tetracycline hydrochloride and antibacterial performance.

In this work, NiFe2O4/g-C3N4 heterostructure was prepared and used for the photocatalytic decomposition of tetracycline hydrochloride antibiotic and for inactivation of E. coli bacteria. The fabricated NiFe2O4/g-C3N4 composite displayed enhanced ability for photodegradation of organic pollutants and disinfection activities compared to the bare samples, because of the enhancement of visible light absorbance, heterojunction formation and photo-Fenton process. The optimized sample 10%-NiFe2O4/g-C3N4 has photodegraded 94.5% of tetracycline hydrochloride in 80 min. The active species trapping experiments revels that ·O2-, h+ and •OH are key decomposing species participated in the antibiotic degradation. It is hoped that the present study will provide a better understanding to fabricate efficient photocatalysts for the decomposition of organic pollutants and disinfection of bacteria.

Experimental Verification of Dielectric Models with a Capacitive Wheatstone Bridge Biosensor for Living Cells: E. coli.

Detection of bioparticles is of great importance in electrophoresis, identification of biomass sources, food and water safety, and other areas. It requires a proper model to describe bioparticles' electromagnetic characteristics. A numerical study of Escherichia coli bacteria during their functional activity was carried out by using two different geometrical models for the cells that considered the bacteria as layered ellipsoids and layered spheres. It was concluded that during cell duplication, the change in the dielectric permittivity of the cell is high enough to be measured at radio frequencies of the order of 50 kHz. An experimental setup based on the capacitive Wheatstone bridge was designed to measure relative changes in permittivity during cell division. In this way, the theoretical model was validated by measuring the dielectric permittivity changes in a cell culture of Escherichia coli ATTC 8739 from WDCM 00012 Vitroids. The spheroidal model was confirmed to be more accurate.

A V-Shaped Microcantilever Sensor Based on a Gap Method for Real-Time Detection of E. coli Bacteria.

This paper presents a dynamic-mode microcantilever sensor based on a gap method. The sensor has a V-shaped microcantilever and a fixed structure at a distance of 2 µm from its free end. The microcantilever is excited by applying an ac electric potential (3 Vp) to its piezoelectric pads and vibrates at its fundamental resonant frequency. An independent ac electric potential (200 kHz, 15 Vpp) is applied to the fixed structure. This creates a non-uniform electric field with its maxima at the gap and exerts a dielectrophoresis (DEP) force. The DEP force attracts and adsorbs the E. coli bacteria to the cantilever edge at the gap. The binding of the bacteria to the cantilever creates a shift in the resonant frequency of the microcantilever sensor, which is detected by a laser vibrometer. The real-time detection of E. coli bacteria samples, diluted in distilled water, was performed for concentrations of 105-103 cells/mL and the real-time frequency shifts were -2264.3 to -755 Hz in 4 min, respectively. The tests were expanded to study the effect of the electric potential amplitude (10, 12, 15 Vpp) and higher frequency shifts were observed for higher amplitudes.

Photodynamic effect of light emitting diodes on E. coli and human skin cells induced by a graphene-based ternary composite.

In this paper, the photodynamic effect of a ternary nanocomposite (TiO2-Ag/graphene) on Escherichia coli bacteria and two human cell lines: A375 (melanoma) and HaCaT (keratinocyte) after exposure to different wavelength domains (blue, green or red-Light Emitting Diode, LED) was analyzed. The results obtained through bioassays were correlated with the morphological, structural and spectral data obtained through FT-IR, XPS and UV-Vis spectroscopy, powder X-Ray diffractometry (XRD) and STEM/EDX techniques, leading to conclusions that showed different photodynamic activation mechanisms and effects on bacteria and human cells, depending on the wavelength. The nanocomposite proved a therapeutic potential for blue light-activated antibacterial treatment and revealed a keratinocyte cytotoxic effect under blue and green LEDs. The red light-nanocomposite duo gave a metabolic boost to normal keratinocytes and induced stasis to melanoma cells. The light and nanocomposite combination could be a potential therapy for bacterial keratosis or for skin tumors.

Development of a Hg-free UV light source incorporating a Kr/Br2 gas, and its application for wastewater treatments.

Mercury lamps are typically the major light sources in water treatments. However, the use of mercury has raised some concerns with regard to the Minamata Convention on Mercury. As such, Hg-free microwave discharged electrodeless lamps (MDELs) that incorporate a rare gas and a halogen gas (R/H-MDEL) have been investigated with such Hg-free mixture filler gases as Kr/Cl2, Xe/Cl2, and Kr/Br2 (R/H). Of these, only the Kr/Br2-MDEL lamp is self-ignited at an inner pressure of 15 Torr when irradiated with microwave radiation. Accordingly, a novel Kr/Br2 three-layer MDEL (Kr/Br2-MDEL) photoreactor was fabricated to assess the optimal gas composition and gas pressure toward its performance vis-à-vis the treatment of model wastewaters contaminated with the tartrazine dye in aqueous media and with Escherichia coli (E. coli) bacteria. The extent of degradation of the tartrazine dye and sterilization of E. coli increased with irradiation time, with microwave radiation power (100, 200, and 300 W), and with increased sample flow rate 0.4 L min‒1 to 0.8 L min‒1. The tartrazine-contaminated wastewater was treated at a flow rate of 0.4 L min‒1 for 60 min of microwave irradiation by three different protocols that resulted in UV (62%) >> UV/ROS (24%) > ROS (0%); ROS denotes reactive oxygen species. After 5 min irradiation of the E.coli wastewater, also at 0.4 L min‒1, the order was UV (99.5%) ≈ UV/ROS (99.3%) >> ROS (14.5%). For comparison, the photosterilization of E. coli with an equivalent Hg/Ar-MDEL light source was also nearly complete (99.7%). Thus, the suitability of the environmentally friendlier Kr/Br2 gas fill to replace Hg/Ar filler gas in MDELs for the photoelimination of organic pollutants and microbial disinfection in aqueous media has been demonstrated.

Co2+-Doped BiOBrxCl1-x hierarchical microspheres display enhanced visible-light photocatalytic performance in the degradation of rhodamine B and antibiotics and the inactivation of E. coli.

In this article, we have synthesized Co2+-doped BiOBrxCl1-x hierarchical nanostructured microspheres, featuring different degrees of Co2+ doping, displaying excellent photocatalytic performance. X-ray diffraction and Raman spectroscopy indicated that the Co2+ ions were successfully doped into the BiOBrxCl1-x nanocrystals. The photodegradation rate of rhodamine B mediated by a doped BiOBrxCl1-x was 150 % greater than that of the non-doped BiOBr. We ascribe the improved photocatalytic capability of the Co2+-doped BiOBrxCl1-x to a combination of its superior degree of light absorption, more efficient carrier separation, and faster interfacial charge migration. The major active species involved in the photodegradation of RhB also has been investigated. Moreover, the doped BiOBrxCl1-x possessed excellent cellular biocompatibility and displayed remarkable performance in the photocatalytic bacterial inactivation.

Cell surface-expression of the phosphate-binding protein PstS: System development, characterization, and evaluation for phosphorus removal and recovery.

Simultaneous overabundance and scarcity of inorganic phosphate (Pi) is a critical issue driving the development of innovative water/wastewater treatment technologies that not only facilitate Pi removal to prevent eutrophication, but also recover Pi for agricultural reuse. Here, a cell-surface expressed high-affinity phosphate binding protein (PstS) system was developed, and its Pi capture and release potential was evaluated. E. coli was genetically modified to express PstS on its outer membrane using the ice nucleation protein (INP) as an anchoring motif. Verification of protein expression and localization were performed utilizing SDS-polyacrylamide gel electrophoresis (SDS-PAGE), western blot, and outer membrane separation analyses. Cell surface characterization was investigated through acid-base titration, X-ray photoelectron spectroscopy (XPS), and Fourier transform infrared spectroscopy (FTIR). These tests provided information on the macromolecular structure and composition of the bacteria surface as well as the proton-exchange properties of the surface functional groups (i.e., pKa values). Phosphate desorption and adsorption batch experiments were conducted to evaluate the effects of temperature, pH, and ionic strength on phosphate capture and release. The PstS surface-displayed cells demonstrated greater potential to release and capture phosphate compared to non-modified cells. Higher temperatures up to 40°C, basic pH conditions (pH = 10.5), and higher ionic strength up to 1.0 mol/L KCl promoted 20%-50% higher phosphate release.

Production and Purification of Recombinant Toxins.

Recombinant expression of toxins enables us to produce adequate quantities of these proteins which can be used to perform experiments at molecular, cellular, and behavioral levels. Furthermore, toxins can be edited by using simple molecular biology methods when producing them recombinantly. Thus, in many cases establishing a protocol for the recombinant expression of a toxin of interest is crucial in exploring the structure and function of the toxin and its effectors. To date, Escherichia coli (E. coli) represents the most widely used heterologous expression system in which recombinant proteins are usually accumulated in the bacterium cytoplasm. However, as many animal toxins contain disulfide bonds they tend to be misfolded and aggregate when found in the reducing E. coli cytoplasm. In contrast, conditions in the bacterium periplasm allow disulfide bond formation and correct folding of such toxins. Here, we describe a protocol for the production and purification of bioactive recombinant disulfide-rich toxins via periplasmic expression.

Self-Organization of the Biological Evolution.

We report here experiments carried out with nonpathogenic Escherichia coli bacterial strains and their phages. This research yielded interesting insights into their activities, occasionally producing genetic variants of different types. In order to not interfere with the genetic stability of the parental strains involved, we found that the bacteria are genetically equipped to only rarely produce a genetic variant, which may occur by a number of different approaches. On the one hand, the genes of relevance for the production of specific genetic variants are relatively rarely expressed. On the other hand, other gene products act as moderators of the frequencies that produce genetic variants. We call the genes producing genetic variants and those moderating the frequencies of genetic variation "evolution genes". Their products are generally not required for daily bacterial life. We can, therefore, conclude that the bacterial genome has a duality. Some of the bacterial enzymes involved in biological evolution have become useful tools (e.g., restriction endonucleases) for molecular genetic research involving the genetic set-up of any living organism.

Nitroxide spin-labeled peptides for DNP-NMR in-cell studies.

Antimicrobial peptides (AMPs) that target lipid membranes show promise as alternatives to conventional antibiotics. However, the molecular mechanisms of membrane perturbation, as most studies are performed in model systems and in-cell structural studies, have yet to be achieved. Solid-state NMR spectroscopy is a valuable technique to investigate peptide-membrane interactions and to determine the structure of peptides, but the short lifespan of bacteria, especially under magic angle spinning conditions, has not permitted in-cell structural studies. Here, we present the first dynamic nuclear polarization (DNP)-NMR in-cell studies of Escherichia coli bacteria incubated with the AMP maculatin 1.1 (Mac1) in combination with novel nitroxide spin-labeled peptides 2,2,6,6-tetramethylpiperidine-N-oxyl-4-amino-4-carboxylic acid (TOAC)-[F3W]-Mac1 (MacW) and TOAC-TOAC-MacW. The in-cell 13C and 15N signal NMR enhancements, and 1H spin-lattice T1 relaxation times showed that TOAC-MacW and TOAC-TOAC-MacW performed better than the more hydrophilic biradical AMUPol used for DNP studies. Furthermore, the pores formed by the AMP increased the signal enhancements and decreased T1 values of specifically 13C- and 15N-labeled Mac1. This approach has a great potential for determining the first in situ structures of AMPs in bacteria.-Sani, M.-A., Zhu, S., Hofferek, V., Separovic, F. Nitroxide spin-labeled peptides for DNP-NMR in-cell studies.

Effect of GO on bacterial cells: Role of the medium type and electrostatic interactions.

Graphene and graphene oxide (GO) both being two-dimensional materials are gaining popularity among researchers as a promising nanomaterial for various medical and biological applications. The aim of this study is to elucidate the influence of nanostructured GO sheets on viability of a model species of gram-negative E. coli bacteria transformed with pRSET-emGFP plasmid in in vitro experiments. It was shown that GO at concentrations between 0.0025 and 2.5 g/l in growth medium inhibits growth of bacterial colonies, while in physiological saline solution (PS) this effect decreases dramatically to the point of complete disappearance. It was shown that in order to obtain a pronounced antibacterial effect one needs to introduce high concentrations of GO into the media (up to 2.5 g/l), which can be important for development of antibacterial materials for biomedical applications. Some of the obtained data provide clear evidence to electrostatic nature of interaction between bacterial and GO sheets. A number of previous papers suggested the process of biofilms formation by bacteria as the primary reason for aggregation between graphene-like materials and bacterial cells. However, formation of flocculent structures consisting of GO and dead bacteria and accompanied with decrease in zeta-potential of particles in the suspension to 18 mV proves that electrostatic interactions play the major role in aggregation. The obtained data can be used for employing GO and similar materials in new systems for water-purification from biological contaminants. Besides, our results stress the importance of accounting for the conditions in which goods and coatings containing graphene-like materials as an antibacterial agent are used, as well as unification of the experimental conditions.

Visual Sensor for Sterilization of Polymer Fixtures Using Embedded Mesoporous Silicon Photonic Crystals.

A porous photonic crystal is integrated with a plastic medical fixture (IV connector hub) to provide a visual colorimetric sensor to indicate the presence or absence of alcohol used to sterilize the fixture. The photonic crystal is prepared in porous silicon (pSi) by electrochemical anodization of single crystal silicon, and the porosity and the stop band of the material is engineered such that the integrated device visibly changes color (green to red or blue to green) when infiltrated with alcohol. Two types of self-reporting devices are prepared and their performance compared: the first type involves heat-assisted fusion of a freestanding pSi photonic crystal to the connector end of a preformed polycarbonate hub, forming a composite where the unfilled portion of the pSi film acts as the sensor; the second involves generation of an all-polymer replica of the pSi photonic crystal by complete thermal infiltration of the pSi film and subsequent chemical dissolution of the pSi portion. Both types of sensors visibly change color when wetted with alcohol, and the color reverts to the original upon evaporation of the liquid. The sensor performance is verified using E. coli-infected samples.

Avoidance of voiding cystourethrography in infants younger than 3 months with Escherichia coli urinary tract infection and normal renal ultrasound.

BACKGROUND AND OBJECTIVE: Urinary tract infection (UTI) represents the most common bacterial infection in infants, and its prevalence increases with the presence of high-grade vesicoureteral reflux (VUR). However, voiding cystourethrography (VCUG) is invasive, and its indication in infants <3 months is not yet defined. This study aims to investigate, in infants aged 0-3 months, if the presence of Escherichia coli versus non-E. coli bacteria and/or normal or abnormal renal ultrasound (US) could avoid the use of VCUG. METHOD: One hundred and twenty-two infants with a first febrile UTI were enrolled. High-grade VUR was defined by the presence of VUR grade ≥III. The presence of high-grade VUR was recorded using VCUG, and correlated with the presence of E. coli/non-E. coli UTI and with the presence of normal/abnormal renal US. The Bayes theorem was used to calculate pretest and post-test probability. RESULTS: The probability of high-grade VUR was 3% in the presence of urinary E. coli infection. Adding a normal renal US finding decreased this probability to 1%. However, in the presence of non-E. coli bacteria, the probability of high-grade VUR was 26%, and adding an abnormal US finding increased further this probability to 55%. CONCLUSIONS: In infants aged 0-3 months with a first febrile UTI, the presence of E. coli and normal renal US findings allow to safely avoid VCUG. Performing VCUG only in infants with UTI secondary to non-E. coli bacteria and/or abnormal US would save many unnecessary invasive procedures, limit radiation exposure, with a very low risk (<1%) of missing a high-grade VUR.