Low Ionic Strength Research Articles

Unusual gold nanoparticle-antibody interactions

The formation of an antibody (Ab) protein corona surrounding gold nanoparticles (AuNPs) is a crucial step in the design of immunological assays. The Ab corona stabilizes AuNPs, preventing their aggregation even at high ionic strength, and can be achieved by simply mixing Abs and AuNPs. In this paper, we report the unusual interactions between AuNPs and the antibody against L1 Cell Adhesion Molecule (L1CAM) purified from rabbits. We have observed that at low ionic strength, the addition of a wide range of concentrations of rabbit monoclonal Abs against L1CAM protein immediately causes the coagulation of citrate-capped gold nanoparticles. This finding is surprising since the addition of proteins to colloidal gold usually forms a stable protein corona. The combination of extinction spectra, dynamic light scattering (DLS), and transmission electron microscopy (TEM) measurements reveals the presence of small clusters of AuNPs coated by the antibodies, as well as micron-sized antibody aggregates. Furthermore, static light scattering measurements demonstrate that Ab self-interactions are attractive (with a negative second virial coefficient, B2) and induce very slow Ab self-aggregation over several months. Overall, these results indicate that, at low ionic strength, the presence of AuNPs enhances Ab-Ab interactions, leading to their rapid aggregation. Simultaneously, the self-aggregation of the antibodies coating the AuNPs results in the formation of nanoparticle clusters. The addition of NaCl to increase the ionic strength fully reverses the coagulation of AuNPs (the Ab-coated AuNPs repel each other) and dissolves the Ab aggregates (the Ab interactions become repulsive, with a positive B2). The AuNPs-induced enhancement of the aggregation process can be explained by considering that the highly favorable binding of Abs on the gold surface compensates for the entropic penalty associated with Ab-Ab aggregation. The phenomenon we observed is specific to anti-L1CAM purified from rabbits and aligns with very old reports on AuNP coagulation induced specifically by the immunoglobulins present in the cerebrospinal fluid of patients suffering from neurosyphilis or multiple sclerosis (C. Lange Zeitschr. Chemotherap., 1912, 1, 44). It is reasonable to hypothesize that other antibodies exhibit this unusual behavior, so this work may aid in the interpretation of “anomalous” results that might otherwise be attributed to errors in fine-tuning AuNPs-Abs conjugation protocols.

Natural organic matter and ionic strength (CaCl2) affect transport, retention and remobilization of silica encapsulated DNA colloids (DNAcol) in saturated sand columns

In the terrestrial environment, interactions between natural organic matter (NOM) and colloids can lead to the formation of an environmental corona around colloids, influencing their transport behaviour and, ultimately, their ecotoxicity. We used a synthetically designed colloid tagged with DNA (DNAcol) as a surrogate for natural colloids and investigated its transport in saturated sand columns. We varied the concentrations of NOM and ionic strength (CaCl2), to better understand the transport and release of DNAcol in porous media under both steady and transient porewater chemistry conditions. In addition, we aimed to understand the main factors that control deposition and release of DNAcol under tested conditions. To induce transient chemistry, we replaced the injection solution containing NOM and/or CaCl2 with Milli-Q water. The results showed that the deposition rate of DNAcol was inversely proportional to the concentration of NOM. The deposition rate increased significantly even under low ionic strength (CaCl2) conditions of tested conditions. Notably, the influence of NOM on the transport of DNAcol was most pronounced at the lowest range of [Ca2+]/DOC ratios, and the attachment of DNAcol to the sand grains was negligible. Moreover, the results showed while the DLVO theory captured the general trend of experimental results, it significantly underestimated the deposition of DNAcol in the presence of CaCl2. Under transient porewater chemistry conditions, colloid remobilization was observed upon flushing the column with Milli-Q water, leading to a secondary peak in the breakthrough curves. We observed that under transient porewater chemistry conditions, when the ionic strength of the solution was 10 mM, the magnitude of the remobilization peak was more significant compared to conditions with 1 mM ionic strength. Our work emphasized the complex interplay between water quality on the one hand and deposition and release of colloidal matter in saturated porous media on the other hand.

Role of Domain-Domain Interactions on the Self-Association and Physical Stability of Monoclonal Antibodies: Effect of pH and Salt.

Monoclonal antibodies (mAbs) make up a major class of biotherapeutics with a wide range of clinical applications. Their physical stability can be affected by various environmental factors. For instance, an acidic pH can be encountered during different stages of the mAb manufacturing process, including purification and storage. Therefore, understanding the behavior of flexible mAb molecules in acidic solution environments will benefit the development of stable mAb products. This study used small-angle X-ray scattering (SAXS) and complementary biophysical characterization techniques to investigate the conformational flexibility and protein-protein interactions (PPI) of a model mAb molecule under near-neutral and acidic conditions. The study also characterized the interactions between Fab and Fc fragments under the same buffer conditions to identify domain-domain interactions. The results suggest that solution pH significantly influences mAb flexibility and thus could help mAbs remain physically stable by maximizing local electrostatic repulsions when mAbs become crowded in solution. Under acidic buffer conditions, both Fab and Fc contribute to the repulsive PPI observed among the full mAb at a low ionic strength. However, as ionic strength increases, hydrophobic interactions lead to the self-association of Fc fragments and, subsequently, could affect the aggregation state of the mAb.

A new method for reactive constant pH simulations.

We present a simulation method that allows us to calculate the titration curves for systems undergoing protonation/deprotonation reactions-such as charged colloidal suspensions with acidic/basic surface groups, polyelectrolytes, polyampholytes, and proteins. The new approach allows us to simultaneously obtain titration curves both for systems in contact with salt and acid reservoir (semi-grand canonical ensemble) and for isolated suspensions (canonical ensemble). To treat the electrostatic interactions, we present a new method based on Ewald summation-which accounts for the existence of both Bethe and Donnan potentials within the simulation cell. We show that the Donnan potential dramatically affects the pH of a suspension. Counterintuitively, we find that in suspensions with a large volume fraction of nanoparticles and low ionic strength, the number of deprotonated groups can be 100% larger in an isolated system, compared to a system connected to a reservoir by a semi-permeable membrane-both systems being at exactly the same pH.

Influence of pH and ionic strength on the physicochemical and structural properties of soybean β-conglycinin subunits in aqueous dispersions

Understanding the impact of pH and ionic strength on the physicochemical and structural properties of soy proteins at subunit level is essential for design and fabrication of many plant-based foods. In this study, soybean β-conglycinin and its subunit fractions αα′ and β were dispersed in solutions with different pH values (3.7, 7.6, and 9.0) at low (5 mM NaCl) and high (400 mM NaCl) ionic strengths, respectively. The solubility, rheology, particle size, zeta potential, microstructure, secondary structure, and tertiary structure of the different dispersions were analyzed using a range of analytical methods. The β-conglycinin, αα′- and β-subunits aggregated near the isoelectric point (pH 3.7). Increasing the ionic strength led to the assembly of more homogeneous units. An increase in ionic strength at pH 7.6 and pH 9.0 led to electrostatic screening, which promoted dissociation of the aggregates. The β-subunit showed a greater sensitivity to pH and ionic strength than the αα′-subunits. Based on the evidence from a range of analytical methods, the highly hydrophilic extension region of the αα′-subunits played an important role in determining the stability of the β-conglycinin dispersions under different environmental conditions. Moreover, the N-linked glycans appeared to impact the conformation and aggregation state of the β-conglycinin.

Biological and physico-chemical characterization of human norovirus-like particles under various environmental conditions

Human noroviruses (HuNoVs) are the predominant etiological agent of viral gastroenteritis in all age groups worldwide. Mutations over the years have affected noroviruses' responses to environmental conditions due to the arrangement of amino acid residues exposed on the VP1 capsid surface of each strain. The GII.4 HuNoV genotype has been the predominant variant for decades, while the GII.17 genotype has often been detected in East Asia since 2014. Here, GII.17 and GII.4 baculovirus-expressed VLPs (virus-like particles) were used to study the biological (binding to HuNoV ligand, namely the ABO and Lewis antigens) and physicochemical properties (size, morphology, and charge) of the HuNoV capsid under different conditions (temperature, pH, and ionic strength). GII.17 showed stability at low and high ionic strength, while GII.4 aggregated at an ionic strength of 10 mM. The nature of the buffers influences the morphology and stability of the VLPs. Here, both VLPs were highly stable from pH 7–8.5 at 25 °C. VLPs retained HBGA binding capability for the pH, ionic strength and temperature encountered in the stomach (fed state) and the small intestine. Increasing the temperature to above 65 °C altered the morphology of VLPs, causing aggregation, and decreased their affinity to HBGAs. Comparing both isolates, GII.17 showed a better stability profile and higher affinity to HBGAs than GII.4, making them interesting candidate particles for a future norovirus vaccine. Biological and physicochemical studies of VLPs are as pertinent as ever in view of the future arrival of VLP-based HuNoV vaccines.

Highly Effective and Efficient Self-Assembled Multilayer-Based Electrode Passivation for Operationally Stable and Reproducible Electrolyte-Gated Transistor Biosensors.

To ensure the operational stability of transistor-based biosensors in aqueous electrolytes during multiple measurements, effective electrode passivation is crucially important for reliable and reproducible device performances. This paper presents a highly effective and efficient electrode passivation method using a facile solution-processed self-assembled multilayer (SAML) with excellent insulation property to achieve operational stability and reproducibility of electrolyte-gated transistor (EGT) biosensors. The SAML is created by the consecutive self-assembly of three different molecular layers of 1,10-decanedithiol, vinyl-polyhedral oligomeric silsesquioxane, and 1-octadecanethiol. This passivation enables EGT to operate stably in phosphate-buffered saline (PBS) during repeated measurements over multiple cycles without short-circuiting. The SAML-passivated EGT biosensor is fabricated with a solution-processed In2O3 thin film as an amorphous oxide semiconductor working both as a semiconducting channel in the transistor and as a functionalizable biological interface for a bioreceptor. The SAML-passivated EGT including In2O3 thin film is demonstrated for the detection of Tau protein as a biomarker of Alzheimer's disease while employing a Tau-specific DNA aptamer as a bioreceptor and a PBS solution with a low ionic strength to diminish the charge-screening (Debye length) effect. The SAML-passivated EGT biosensor functionalized with the Tau-specific DNA aptamer exhibits ultrasensitive, quantitative, and reliable detection of Tau protein from 1 × 10-15 to 1 × 10-10 M, covering a much larger range than clinical needs, via changes in different transistor parameters. Therefore, the SAML-based passivation method can be effectively and efficiently utilized for operationally stable and reproducible transistor-based biosensors. Furthermore, this presented strategy can be extensively adapted for advanced biomedical devices and bioelectronics in aqueous or physiological environments.

Graphene versus concentrated aqueous electrolytes: the role of the electrochemical double layer in determining the screening length of an electrolyte

Understanding the performance of graphene devices in contact with highly concentrated aqueous electrolytes is key to integrating graphene into next-generation devices operating in sea water environments, biosensors, and high-density energy production/storage units. Despite significant efforts toward interpreting the structure of the electrochemical double layer at high concentrations, the interface between graphene-based materials and concentrated aqueous solutions has remained vaguely described. In this study, we demonstrate the use of graphene-based chemiresistors as a technique to indirectly quantify the experimental screening length of concentrated electrolytes that could clarify the interpretation of electrochemical measurements conducted at low ionic strength. We report a breakdown of the Debye–Hückel theory in the proximity of graphene surfaces at lower concentrations (10–50 mM) than previously reported for other systems, depending on cation size, dissolved oxygen concentration, and degree of graphene defectivity.

Enhanced adsorption of phenol from aqueous solution by KOH combined Fe-Zn bimetallic oxide co-pyrolysis biochar: Fabrication, performance, and mechanism

In this study, impregnation combined with KOH activation with different mixing methods was used to prepare magnetic biochar. The effects of synthetic method on biochar physicochemical properties and adsorption performance were explored. The results showed that treatment of a Fe-Zn oxide with KOH activation provided excellent adsorption properties with adsorption capacity of 458.90 mg/g due to well-developed microporous structure and rich-in O-containing functional groups as well as exposed oxidizing functional groups (Fe2O3 and FeOOH). Langmuir-Freundlich and pseudo-second-order models accurately fit phenol adsorption. Neutral conditions (pH = 6) and lower ionic strengths were beneficial to phenol removal. Additionally, the predominant adsorption processes were physisorption and chemisorption. Correlation analyses and characterization data confirmed that pore filling, π-π interactions and surface complexation were the dominant driving forces for phenol adsorption. This research provides an environmentally friendly method for utilizing agricultural wastes for the removal of a variety of pollutions from aquatic environment.

Synchrotron X-ray study of intrinsically disordered and polyampholytic Tau 4RS and 4RL under controlled ionic strength.

Aggregated and hyperphosphorylated Tau is one of the pathological hallmarks of Alzheimer's disease. Tau is a polyampholytic and intrinsically disordered protein (IDP). In this paper, we present for the first time experimental results on the ionic strength dependence of the radius of gyration (Rg) of human Tau 4RS and 4RL isoforms. Synchrotron X-ray scattering revealed that 4RS Rg is regulated from 65.4 to 58.5Å and 4RL Rg is regulated from 70.9 to 57.9Å by varying ionic strength from 0.01 to 0.592M. The Rg of 4RL Tau is larger than 4RS at lower ionic strength. This result provides an insight into the ion-responsive nature of intrinsically disordered and polyampholytic Tau, and can be implicated to the further study of Tau-Tau and Tau-tubulin intermolecular structure in ionic environments.

Next-Generation Nanopore Sensors Based on Conductive Pulse Sensing for Enhanced Detection of Nanoparticles.

Nanopore sensing has been successfully used to characterize biological molecules with single-molecule resolution based on the resistive pulse sensing approach. However, its use in nanoparticle characterization has been constrained by the need to tailor the nanopore aperture size to the size of the analyte, precluding the analysis of heterogeneous samples. Additionally, nanopore sensors often require the use of high salt concentrations to improve the signal-to-noise ratio, which further limits their ability to study a wide range of nanoparticles that are unstable at high ionic strength. Here, a new paradigm in nanopore research that takes advantage of a polymer electrolyte system to comprise a conductive pulse sensing approach is presented. A finite element model is developed to explain the conductive pulse signals observed and compare these results with experiments. This system enables the analytical characterization of heterogeneous nanoparticle mixtures at low ionic strength . Furthermore, the wide applicability of the method is demonstrated by characterizing metallic nanospheres of varied sizes, plasmonic nanostars with various degrees of branching, and protein-based spherical nucleic acids with different oligonucleotide loadings. This system will complement the toolbox of nanomaterials characterization techniques to enable real-time optimization workflow for engineering a wide range of nanomaterials.

(Invited) Voltammetric Examination of Carbon Nanoelectrodes Under Mixed Diffusion - Migration Mass Transport Conditions

In general, experiments carried out with nanoelectrodes are similar to those with microelectrodes. However, the extremely increased efficiency of the mass transport to nanoelectrodes results in steady-state current conditions in ultra-short times and high current densities. The transport of electroactive species to the working electrode surface is mainly realized by diffusion. The contribution of migration to the total mass transport is usually eliminated by shortening the thickness of the electrical double layer (EDL) achieved by the addition of supporting electrolyte excess. This simplifies the theoretical description of mass transport to pure diffusion but on the other hand, modifies physicochemical properties, including transport parameters of the analyzed environment. In systems containing a deficiency of supporting electrolyte, migration contributes to mass transport. The transport studies in these systems require the employment of small-sized electrodes (micro- / nanoelectrodes) and potentiostats capable of measuring ultralow currents.Migration introduces an additional variable (electrostatic potential) to the transport expressed by the Nernst-Planck equation. Under the low ionic strength conditions, the transport dynamics depends on the concentration profiles of a substrate, product, cations and anions of supporting electrolyte, and an uncompensated electrostatic potential gradient. In practice, the contribution of migration can be quantitatively related to the support ratio parameter (ξ), defined as the ratio of the concentrations of electroinactive charge carriers (i.e. not involved in the electrode reaction) to the concentration of the electrode reaction substrate. In terms of the ξ parameter, migration has a negligible effect on mass transport under the conditions of high values of the support ratio (ξ > 100). As the support ratio decreases (ξ < 1) the potential gradient occurs due to increased resistivity of the system and, consequently, the total mass transport is affected by the migrational contribution.The problem of diffusion-migration transport in voltammetric experiments involving microelectrodes was treated theoretically and the derived models allow one to predict the voltammograms for various types of electrode processes carried out under the conditions of various concentration of supporting electrolyte. The major factor complicating the theoretical description of mass transport induced by migration is the influence of an EDL. In the system containing a deficiency of supporting electrolyte, EDL may occupy a significant part of the diffusion layers of the substrate and the product of the electrode process. With the decrease of the electrode radius (below 100 nm), the EDL start to overlap with the diffusion layer and significantly affects the transport even in the system containing substantial concentrations of supporting electrolyte.To explore new research domains for nanoelectrodes, the procedures for the fabrication of nanoelectrodes of reproducible parameters and the theoretical description of the mass transport to nanoelectrodes under mixed diffusion-migration transport conditions have to be developed.The ultrasmall electrodes are highly susceptible to electrostatic mechanical and electrochemical damage, leading to the formation of recessed or protruding electrodes with altered current response, which is an additional impediment in the studies with their use. As a result of the damage causing recession, the electrodes lose their current response and give low feedback in their use as probes in scanning electrochemical microscopy (SECM). The damage caused by electrostatic discharging can be avoided by handling and storing the electrodes grounded together with objects in the vicinity.This work demonstrates the voltammetric behavior of the fabricated carbon nanoelectrodes in the presence and absence of supporting electrolyte. The ferrocene derivatives of various charges served as redox probes for the studies of diffusion-migration transport to nanoelectrodes. The obtained results are confronted to the theoretical predictions derived for microelectrodes. They revealed that migration of electroactive species to nanoelectrodes does not introduce significant changes to voltammetric signal as much as for bigger microelectrodes. Moreover, the elimination of supporting electrolyte in most cases led to the poor development of the voltammetric waves at nanoelectrodes. The possible reason may be related to the breakdown of the electroneutrality in the region adjacent to the nanoelectrode, since the electrical double layer for nanoelectrodes may occupy a significant fraction of the transport depletion layer. The uncertainty in the concentrations of ionic impurities (that determine the actual support ratio) in the layer adjacent to the electrode is another factor that may produce unpredictable and, to some extent, random voltammetric behavior of charged redox probes at nanoelectrodes in the absence of supporting electrolyte.

Effect of montmorillonite and humic acid on cotransport of g-C3N4 and lead: The role of triazine ring in lead adsorption and deposition in soil components

In the past few years, Graphitic carbon nitride (g-C3N4) has been produced on a large scale and has been widely used because of its excellent optical properties and physicochemical stability. It is easy to ignore the risk of g-C3N4 transport in the environment, including soil and groundwater, due to the non-toxicity of metals. Based on this situation, we investigated the effects of pumping flow rate, ionic strength, and the presence of lead (Pb) on the transport behavior of g-C3N4 in quartz sand, montmorillonite, and humic acid-coated sand through column experiments. The retention of g-C3N4 in the media and the adsorption mechanism of Pb onto g-C3N4 were analyzed by FTIR and XPS. The results showed that fast flow rate and low ionic strength were favorable conditions for the transport of g-C3N4. The cotransport experiment results showed that Pb reduced the outflow of g-C3N4 by more than 50%. Contrarily, 6.95%, 0.43%, and 11.01% Pb were hindered in quartz sand, montmorillonite, and humic acid-coated sand due to the presence of g-C3N4. Compared with the uncontaminated media, the recoveries of g-C3N4 in Pb contaminated quartz sand, montmorillonite, and humic acid-coated sand were reduced by 0.34%, 5.86%, and 15.34%, respectively. Concomitantly, which were attributed to the special triazine ring structure and the abundant bonding modes with soil components and Pb, g-C3N4 particles were easier to deposit in montmorillonite and humic acid-coated sand, and could be used as the carrier to release 6.70% and 5.64% Pb from contaminated quartz sand and humic acid-coated sand.

Serum Albumin Aggregation Facilitated by Cobalt and Chromium Metal Ions.

The interaction of serum proteins with cobalt (Co) and chromium (Cr) ions is poorly understood, but it is suspected to result in protein aggregation, which may alter the corrosion process of biomedical CoCr alloys or result in adverse health effects. Here, we study the aggregation ability and mechanism of bovine serum albumin (BSA) induced or accelerated by aqueous Co(II) and Cr(III) ions. The metal salts were selected by chemical speciation modeling, and they did not affect the pH or precipitate under simulated physiological conditions (150 mM NaCl and pH 7.3). The counterion of Cr(III) influenced the binding to BSA only at physiologically irrelevant low ionic strength. This study used a variety of spectroscopic and light scattering methods. It was determined that both metal ions and an equimolar mixture of metal ions have the potential to induce protein aggregation. Melting curves collected by circular dichroism spectroscopy indicate that Co(II) significantly reduced BSA's melting temperature when compared with Cr(III) or an equimolar mixture of Co(II) and Cr(III), both of which increased the melting temperature of BSA. The metal ions in solution preferentially interacted with BSA, resulting in the depletion of metal ions from the surrounding protein-free solution. Finally, this study suggests that the likely mechanism for Co(II)- and Cr(III)-induced BSA aggregation is salt bridging between protein molecules.

Exploring the Influence of Chemical Conditions on Nanoparticle Graphene Oxide Adsorption onto Clay Minerals.

High concentrations of graphene oxide (GO), a nanoparticle substance with rapid manufacturing development, have the ability to penetrate the soil surface down to the mineral-rich subsurface layers. The destiny and distribution of such an unusual sort of nanomaterial in the environment must therefore be fully understood. However, the way the chemistry of solutions impacts GO nanoparticle adsorption on clay minerals is still unclear. Here, the adsorption of GO on clay minerals (e.g., bentonite and kaolinite) was tested under various chemical conditions (e.g., GO concentration, soil pH, and cation valence). Non-linear Langmuir and Freundlich models have been applied to describe the adsorption isotherm by comparing the amount of adsorbed GO nanoparticle to the concentration at the equilibrium of the solution. Our results showed fondness for GO in bentonite and kaolinite under similar conditions, but the GO nanoparticle adsorption with bentonite was superior to kaolinite, mainly due to its higher surface area and surface charge. We also found that increasing the ionic strength and decreasing the pH increased the adsorption of GO nanoparticles to bentonite and kaolinite, mainly due to the interaction between these clay minerals and GO nanoparticles' surface oxygen functional groups. Experimental data fit well to the non-linear pseudo-second-order kinetic model of Freundlich. The model of the Freundlich isotherm was more fitting at a lower pH and higher ionic strength in the bentonite soil while the lowest R2 value of the Freundlich model was recorded at a higher pH and lower ionic strength in the kaolinite soil. These results improve our understanding of GO behavior in soils by revealing environmental factors influencing GO nanoparticle movement and transmission towards groundwater.

Thermal Gelation of Proteins from Cajanus cajan Influenced by pH and Ionic Strength.

Cajanus cajan [pigeon pea (PP)] is an important legume crop for subsistence agriculture and its seeds are an alternative plant-based protein source. PP protein isolates (PPI) are able to form heat-induced gels that could be used for food applications. The aim of this work was to study the influence of pH (2.1, 3.9, 6.3, and 8.3) and ionic strength (μ) (0.10 and 0.54) on thermal stability and thermal gelation of PPI obtained by alkaline extraction (pH8.0) and isoelectric precipitation. Thermal stability of PPI changed with pH variation at low ionic strength (μ = 0.10), decreasing this dependence with the increase of ionic strength (μ = 0.54). At μ = 0.10, gelation capacity of PPI was lower at pH2.1 and pH3.9. These gels presented a coarse network, which entails low WHC. At pH6.3 and pH8.3, gels showed a solid-like character with a compact and homogeneous matrix, with better WHC. At μ = 0.54, gel formation was favoured at pH2.1 and pH3.9. G'20/G'95 ratio values and differential solubility results suggest that hydrogen bonds and electrostatic interactions could play an important role in gel formation at pH6.3 and pH8.3.

DNA condensation by cysteine containing tetra-arginine ligands as a model for ionizing radiation damage to chromatin

We examined DNA binding and condensation by AcCR4NH2 and CR4NH2 ligands. By this approach, we can model physical DNA damage by ionizing radiation. The binding constant was 1 × 106 L mol−1 at low ionic strength (ca. 10 mmol L−1 sodium ions) and the resulting condensate size was ca. 1 μm. Given the similar base pair concentrations of ca. 250 mmol L−1 in condensed DNA and chromatin, DNA damage yields by ionizing irradiation in condensates with related oligoarginine ligands are comparable to those in eukaryotic nuclei. Therefore, with the benefit of these results, we can envisage an experimental system suitable for modeling DNA damage by ionizing radiation because it reproduces well the essential conditions in cell nuclei that affect this damage, particularly the high concentrations of DNA and protein in chromatin and the resulting highly efficient radical scavenging environment.

Structural insights into anion selectivity and activation mechanism of LRRC8 volume-regulated anion channels

Volume-regulated anion channels (VRACs) are hexamers of LRRC8 proteins that are crucial for cell volume regulation. N termini (NTs) of the obligatory LRRC8A subunit modulate VRACs activation and ion selectivity, but the underlying mechanisms remain poorly understood. Here, we report a 2.8-Å cryo-electron microscopy structure of human LRRC8A that displays well-resolved NTs. Amino-terminal halves of NTs fold back into the pore and constrict the permeation path, thereby determining ion selectivity together with an extracellular selectivity filter with which it works in series. They also interact with pore-surrounding helices and support their compact arrangement. The C-terminal halves of NTs interact with intracellular loops that are crucial for channel activation. Molecular dynamics simulations indicate that low ionic strength increases NT mobility and expands the radial distance between pore-surrounding helices. Our work suggests an unusual pore architecture with two selectivity filters in series and a mechanism for VRAC activation by cell swelling.

Interaction of polystyrene nanoplastics with impurity-bearing ferrihydrite and implication on complex particle sedimentation

Nanoplastics (NPs) usually coexist with impurity-bearing ferrihydrite (ImFh), and their interaction is related to their environmental fate. In this study, the aggregation between ImFh (impurities: Al, Mn and Si) and polystyrene nanoplastics (PSNPs), as well as the sedimentation of ImFh-PSNP complex particles in the aqueous phase were investigated systematically with particle concentrations of 100 mg/L ImFh and 10 mg/L PSNPs. Our results revealed that the PSNP suspension was dispersive and stable under various pH values and low ion strength. After coexisting with ImFh, PSNPs aggregated with the positively charged ImFh to form ImFh-PSNP complex particles, which destroyed the stability of PSNPs. The increase in pH and Na+ concentration could inhibit their aggregation, but high Na+ concentration (>20 mM) caused the homoaggregation of PSNPs. The aggregation capacity of PSNPs with ImFh was in the order of Al-bearing Fh > Fh > Mn-bearing Fh > Si-bearing Fh. Zeta potential and Derjaguin–Landau–Verwey–Overbeek (DLVO) calculations indicated that Al-bearing Fh showed higher positive potential than pure Fh, which caused stronger electrostatic interactions with PSNPs. However, Mn and Si in ImFh decreased the positive potential and inhibited the electrostatic interaction with PSNPs, and the effect of Si was greater than that of Mn. The aggregation between ImFh and PSNPs inhibited the sedimentation of their complex particles, and the higher aggregation capacity appeared to have a greater inhibition degree. Due to the “electrostatic patches” effect of PSNPs, the energy barrier of the ImFh-PSNPs particles was higher than that of the ImFh particles. Our findings clarified the influence of impurities on the interaction between ImFh and PSNPs and provided insight regarding their fate in the environment.

Transcriptomic Analysis Followed by the Isolation of Extracellular Bacteriolytic Proteases from Lysobacter capsici VKM B-2533T.

The aim of the study was to search for, isolate and characterize new bacteriolytic enzymes that show promising potential for their use in medicine, agriculture and veterinary. Using a transcriptomic analysis, we annotated in Lysobacter capsici VKM B-2533T the genes of known bacteriolytic and antifungal enzymes, as well as of antibiotics, whose expression levels increased when cultivated on media conducive to the production of antimicrobial agents. The genes of the secreted putative bacteriolytic proteases were also annotated. Two new bacteriolytic proteases, Serp and Serp3, were isolated and characterized. The maximum bacteriolytic activities of Serp and Serp3 were exhibited at low ionic strength of 10 mM Tris-HCl, and high temperatures of, respectively, 80 °C and 70 °C. The pH optimum for Serp was 8.0; for Serp3, it was slightly acidic, at 6.0. Both enzymes hydrolyzed autoclaved cells of Micrococcus luteus Ac-2230T, Proteus vulgaris H-19, Pseudomonas aeruginosa and Staphylococcus aureus 209P. Serp also digested cells of Bacillus cereus 217. Both enzymes hydrolyzed casein and azofibrin. The newly discovered enzymes are promising for developing proteolytic antimicrobial drugs on their basis.