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To achieve efficient and safe cationic carrier-mediated gene delivery for gene therapy, the optimal ratio of carrier to DNA in formulations is a key factor and it is usually determined prior to transfection experiments. In this work, a simplified drop-and-read assay was developed for the first time using paper as a platform to estimate the equivalence ratio of cationic carriers to negatively charged DNA. By spotting a series of complexes containing varied ratios of carrier to DNA on filter paper, then allowing them to dry and finally dropping yellowish-green anionic 2',7'-dichlorofluorescein dye solution on top of the complexes, the equivalence point was detectable by the instant formation of stable pink spots as a result of the dye adsorption onto the positively charged complexes and free carriers. The method gave the same results as those determined by gel retardation assay and zeta potential measurement, however it allowed more rapid reporting of results in 5 min and required no tedious steps, harmful reagents or expensive instruments. By using paper instead of microcentrifuge tubes and omitting centrifugation, plasticware and electrical energy were no longer consumed and disposal of this degradable material was more environmentally friendly. With respect to analytical performance, filter paper inherently holding negative charge helped to trap and concentrate the complexes on the white background, enabling greater visibility of the colored spots and a lower required amount of DNA used for the assay. The method was successfully applied to estimate the equivalence ratios in a variety of gene delivery formulations containing different types of cationic carriers, i.e. polymers, dendrimers, liposomes and niosomes.

We investigated the effect that two common types of cationic polyelectrolytes used in papermaking might have on the performance of paper diagnostics using blood typing as an example. The results were analyzed in terms of red blood cells (RBC) retention and antibody–antigen specificity. Two questions were addressed: (1) can poly(amido-amine) epichlorohydrin (PAE) typically used for paper wet strength affect the diagnostic performance? (2) can high molecular weight cationic polyacrylamide (CPAM) employed as retention aid enhance or affect the selectivity and sensitivity of paper diagnostics? A series of paper varying in type of fibers and drying process were constructed with PAE and tested for blood typing performance. Residual PAE has no significant effect on blood typing paper diagnostics under normal conditions. Positives are unaffected with PAE, while negatives lose slight sharpness as some RBCs are unselectively retained. CPAM, the most common retention aid, can also be used to retain cells and biomolecules on paper. Paper towel was treated with CPAM solutions varying in polymer concentration and charge density and tested for blood typing. We found that CPAM dried on paper can retain RBC. CPAM affects the negative tests by retaining non-specifically individual RBC on fibers. RBC retention increases non-linearly with the CPAM charge density and concentration. As expected, wet CPAM retain RBCs at concentrations higher than 0.1 wt%. As paper diagnostics are becoming a reality, more realistic papers than the Whatman filter paper will be engineered. This study provides guidance on how best use the required polymeric wet-strength and retention agents.

The use of non-wood fibers in the paper industry has been an economical and environmental necessity. The application of dry-strength agents has been a successful method to enhance the strength properties of paper. The experimental results evidencing the potential of chitosan and cationic starch utilization in bagasse paper subjected to hot water pre-extraction has been presented in this paper. The research analyzes the surface properties alterations due to these dry-strength agents. Inverse gas chromatography was used to evaluate the properties of surface chemistry of the papers namely the surface energy, active sites, surface area as well as the acidic/basic character. The results of the study revealed that the handsheets process causes surface arrangement and orientation of chemical groups, which induce a more hydrophobic and basic surface. The acid-base surface characteristics after the addition of dry-strength agents were the same as the bagasse handsheets with and without hot water pre-extraction. The results showed that the dry-strength agent acts as a protecting film or glaze on the surfaces of bagasse paper handsheets.

This study examines and quantifies the effect of cationic polyelectrolyte adsorption on paper on the aggregation and retention of gold nanoparticles (AuNPs) to optimize their Surface Enhanced Raman Scattering (SERS) enhancement factor and sensitivity. Aggregation of metallic nanoparticles is known as a key factor for intense SERS enhancement. Paper substrates were treated with aqueous solutions of cationic polyacrylamide (CPAM) varying in concentration, charge density, and molecular weight to control the AuNPs' aggregate size distribution and surface coverage on paper. The Raman Enhancement Factor of AuNPs-CPAM paper was almost an order of magnitude greater than for the untreated AuNPs paper. The high loading and uniform distribution of AuNPs aggregates on CPAM pre-treated paper contributed toward the excellent SERS reproducibility, sensitivity, and high enhancement factor. This configuration of AuNP on paper was favoured by treating the substrate with CPAM solutions of higher concentrations, higher charge density, and greater molecular weight.

Chemistry - a European journal 27(48), 12371 - 12379 (2021). doi:10.1002/chem.202101515, We investigated the photoionization and fragmentation of isolated metal protoporphyrin IX cations (MPPIX+ with M=Fe, Co, Zn) by means of vacuum-ultraviolet (VUV) action spectroscopy in the energy range of 8.5���35 eV. Experiments were carried out in the gas phase by interfacing an electrospray ionization tandem mass spectrometer with a synchrotron beamline. The mass spectra and partial ion yields show that photoexcitation of the precursor ions predominantly leads to .CH$_2$COOH radical side-chain losses of the macrocycle with additional methyl radical (.CH$_3$) side-chain losses. Ionization, in contrast, leads to the formation of the intact ionized precursor and various doubly charged fragments which are mostly due to side-chain cleavages. Although statistical fragmentation dominates, we found evidence for non-statistical processes such as new fragments involving for example single and double H2O losses, indicating that different relaxation mechanisms are at play upon photoionization compared to photoexcitation. The measured ionization energies were 9.6��0.2 eV, 9.4��0.2 eV and 9.6��0.2 eV for FePPIX$^+$, CoPPIX$^+$ and ZnPPIX$^+$, respectively., Published by Wiley-VCH, Weinheim

Strongly Lewis acidic cationic aluminium complexes, stabilized by β–diketiminate (BDI) ligands and free of Lewis bases, have been prepared as their B(C6F5)4 − salts and were investigated for catalytic activity in imine hydrogenation. The backbone (R1) and N (R2) substituents on the R1,R2BDI ligand (R1,R2BDI=HC[C(R1)N(R2)]2) influence sterics and Lewis acidity. Ligand bulk increases along the row Me,DIPPBDI, A series of strongly Lewis acidic cationic aluminium complexes, stabilized by β–diketiminate (BDI) ligands and free of Lewis bases, have been prepared as their B(C6F5)4 − salts. Their catalytic activity in imine hydrogenation depends strongly on the substituents in the BDI ligand. DFT calculations on the mechanism reveal two interconnected catalytic cycles. Hydrogen is activated either by Al⋅⋅⋅imine or Al⋅⋅⋅amine Frustrated‐Lewis‐Pairs.

Journal article Cationic polymers with various structures have been widely investigated in the areas of medical diagnostics and molecular, biology because of their unique binding properties and capability to interact with biological molecules in complex biological environments.; In this work-we report the grafting of a linear cationic polymer from an atom transfer radical polymerization (ATRP) initiator bound to cellulose paper surface. We show successful binding of ATRP initiator onto cellulose paper and grafting of polymer chains from the immobilized initiator with ATRP. The cellulose paper grafted polymer was used in combination with PicoGreen (PG) to demonstrate detection of nucleic acids in the nanogram range in homogeneous solution and in a biological sample (serum). The results showed specific identification of hybridized DNA after addition of PG in both solutions. peer-reviewed

The homospermidine synthases from P. aeruginosa and B. viridis, as well as their single-residue variants, are compared based on crystal structures and activity assays. A high structural similarity is demonstrated, suggesting the equivalent involvement of relevant residues in the reaction mechanism and catalytic dependence on cation–π interaction., Polyamines influence medically relevant processes in the opportunistic pathogen Pseudomonas aeruginosa, including virulence, biofilm formation and susceptibility to antibiotics. Although homospermidine synthase (HSS) is part of the polyamine metabolism in various strains of P. aeruginosa, neither its role nor its structure has been examined so far. The reaction mechanism of the nicotinamide adenine dinucleotide (NAD+)-dependent bacterial HSS has previously been characterized based on crystal structures of Blastochloris viridis HSS (BvHSS). This study presents the crystal structure of P. aeruginosa HSS (PaHSS) in complex with its substrate putrescine. A high structural similarity between PaHSS and BvHSS with conservation of the catalytically relevant residues is demonstrated, qualifying BvHSS as a model for mechanistic studies of PaHSS. Following this strategy, crystal structures of single-residue variants of BvHSS are presented together with activity assays of PaHSS, BvHSS and BvHSS variants. For efficient homospermidine production, acidic residues are required at the entrance to the binding pocket (‘ionic slide’) and near the active site (‘inner amino site’) to attract and bind the substrate putrescine via salt bridges. The tryptophan residue at the active site stabilizes cationic reaction components by cation–π interaction, as inferred from the interaction geometry between putrescine and the indole ring plane. Exchange of this tryptophan for other amino acids suggests a distinct catalytic requirement for an aromatic interaction partner with a highly negative electrostatic potential. These findings substantiate the structural and mechanistic knowledge on bacterial HSS, a potential target for antibiotic design.

The effect of polymer on the retention and the thermal stability of bioactive enzymatic papers was measured using a colorimetric technique quantifying the intensity of the enzyme–substrate product complex. Alkaline phosphatase (ALP) was used as model enzyme. Three water soluble polymers: a cationic polyacrylamide (CPAM), an anionic polyacrylic acid (PAA) and a neutral polyethylene oxide (PEO) were selected as retention aids. The model polymers increased the enzyme adsorption on paper by around 50% and prevented enzyme desorption upon rewetting of the papers. The thermal deactivation of ALP retained on paper with polymers follows two sequential first order reactions. This was also observed for ALP simply physisorbed on paper. The retention aid polymers instigated a rapid initial deactivation which significantly decreased the longevity of the enzymatic papers. This suggests some enzyme–polymer interaction probably affecting the enzyme tertiary structure. A deactivation mathematical model predicting the enzymatic paper half-life was developed.

Instituto de Recursos Naturales y Agrobiología, CSIC, P.O. Box 1052, E-41080, Seville, Spain E-mail address: delrio@irnase.csic.es, The composition of pitch deposits occurring in pulp sheets and mill circuits during soda/anthraquinone pulping and elemental chlorine-free pulp bleaching of bast fibers of industrial hemp (Cannabis sativa) has been studied. Pitch deposits were extracted with acetone, and the extracts analyzed by gas chromatography (GC) and gas chromatography/mass spectrometry (GC/MS). Acetone extracts (15-25% of pitch deposits) were constituted by the defoamers used at the mill and by lipophilic extractives from hemp fibers. Acetone-insoluble residues (75-85% of pitch deposits) were analyzed by pyrolysis-GC/MS in the presence and absence of tetramethylammonium hydroxide. These residues were constituted by salts of fatty acids (arising from hemp fibers) with calcium, magnesium, aluminum and other cations that were identified in the deposits. It was concluded that inappropriate use of defoamer together with the presence of multivalent ions seemed to be among the causes of hemp extractives deposition in the pitch problems reported here., This study has been supported by the Spanish Ministerio de Ciencia y Tecnología (MCYT) and FEDER funds (projects 2FD97-0896-C02-02 and AGL2002-00393). A.G. acknowledges a "Ramón y Cajal" contract of the Spanish MCYT. We also thank CELESA (Tortosa, Spain) for providing the samples.

Plastic waste in the ocean and on land in the form of nanoplastics is endangering food and drinking water supplies, raising the need for new strategies for the removal of plastic nanoparticles from complex media. In the present contribution we suggest considering ionic liquids as extractants, since they show several advantageous properties that may facilitate the design of efficient separation processes. Through varying the anion and the side chain at the cation, the interactions between the extractant and the polymer can be strengthened and tuned, and thereby the disintegration of the particle into separate polymer chains can be controlled. Oxidized moieties can also be efficiently solvated, given the amphiphilic nature of the considered ionic liquids, allowing also realistic particles to be extracted into these solvents. The phase transfer was found to be thermodynamically and kinetically possible, which is supported by the complicated structure of the ionic liquid‐water interface through the rearrangement of the interfacial ions, and the formation of a micelle around the plastic already at the edge of the aqueous phase., Hydrophobic ionic liquids are suggested as extracting solvents for the removal of nanoplastics from aqueous systems, to reduce the concentration of these potentially hazardous materials. Based on molecular modeling, comparison with molecular solvents reveal clear advantages for ionic liquids in terms of solvation and disintegration of the plastic particle, and the phase transfer mechanism. The results suggest ways of tuning and improving an extraction process.

Synthesis of mixed‐substituted tetraphosphetanes (RP−PtBu)2 (R=Ph (4), Py (5); Py=2‐pyridyl) is achieved from the condensation of dipyrazolylphosphanes RPpyr2 (R=Py (1), Ph (3); pyr=3,5‐dimethylpyrazolyl) as P1‐building block (R−P) and tBuPH2 in an equimolar ratio. Compound 5 is of special interest since the presence of two pyridyl‐substituents as well as the P4‐core allows for a rich coordination chemistry with coinage metal salts [Cu(MeCN)4][OTf], Ag[OTf] and in situ formed [Au(tht)][OTf] (tht=tetrahydrothiophene). Both tetraphosphetanes undergo alkylation reaction with MeOTf to give a series of tetraphosphetanium and tetraphosphetanediium triflate salts with additional methylation of the pyridyl‐moiety in case of 5 resulting in interesting novel cyclic trications. Harsh reaction condition and an excess of MeOTf converts 5 into the cyclic trication [‐P(MePy)PMe2P(MePy)PtBu‐]3+ (13 3+; MePy=1‐methylpyridiniumyl) through the elimination of isobutene. This salt undergoes a complicated rearrangement reaction involving a P−P/P−P bond metathesis to form trication [‐P(MePy)3PtBu‐]3+ (17 3+) when reacted with Me2PPMe2., Choose your donor: A convenient route towards mixed‐substituted tetraphosphetanes was established, enabling the synthesis of (PhP‐PtBu)2 and (PyP‐PtBu)2 (Py=2‐pyridyl) in good yields. The coordination chemistry of (PyP‐PtBu)2 was investigated by reacting it with coinage metal triflate salts. Further methylation studies on both tetraphosphetanes revealed an interesting chemistry of multiply charged, cylic polyphosphanes.

Liposomes have been widely applied in bioanalytical assays. Most liposomes used bare negative charges to prevent non-specific binding and increase colloidal stability. Here, in contrast, highly stable, positively charged liposomes entrapping the fluorescent dye sulforhodamine B (SRB) were developed to serve as a secondary, non-specific label‚ and signal amplification tool in bioanalytical systems by exploiting their electrostatic interaction with negatively charged vesicles, surfaces, and microorganisms. The cationic liposomes were optimized for long-term stability (> 5 months) and high dye entrapment yield. Their capability as secondary, non-specific labels was first successfully proven through electrostatic interactions of cationic and anionic liposomes using dynamic light scattering, and then in a bioassay with fluorescence detection leading to an enhancement factor of 8.5 without any additional surface blocking steps. Moreover, the cationic liposomes bound efficiently to anionic magnetic beads were stable throughout magnetic separation procedures and could hence serve directly as labels in magnetic separation and purification strategies. Finally, the electrostatic interaction was exploited for the direct, simple, non-specific labeling of gram-negative bacteria. Isolated Escherichia coli cells were chosen as models and direct detection was demonstrated via fluorescent and chemiluminescent liposomes. Thus, these cationic liposomes can be used as generic labels for the development of ultrasensitive bioassays based on electrostatic interaction without the need for additional expensive recognition units like antibodies, where desired specificity is already afforded through other strategies. Graphical abstract Electronic supplementary material The online version of this article (10.1007/s00216-020-02612-w) contains supplementary material, which is available to authorized users.

Bioflocculant-producing bacteria were isolated from various water reservoirs and sediments of the water treatment plant. Four promising strains were identified by standard biochemical methods and 16s rRNA gene sequencing. Bioflocculants were produced in a batch bioreactor of 3 L under optimized conditions. Fourier transformed infrared spectroscopy and scanning electron microscopy (SEM) were used to confirm the chemical and morphological nature of bioflocculants. Anionic and cationic textile dyes congo red (CR) and rhodamine-B (RB) decolourization efficiency by ethanol precipitated bioflocculants were accessed under different values of pH, temperature, dose of flocculant and presence of monovalent, divalent and trivalent cations. Bioflocculants of all the four isolates were found to be highly efficient in decolourization of dye from an aqueous medium with the removal rate up to 99.56%. The removal rate of CR and RB from aqueous medium was largely influenced by the physiochemical condition of the solution viz. pH, temperature, concentration of ions and dose of flocculants. The microbial bioflocculants are biodegradable and highly stable as well as possess abroad range of pH, temperature and ions tolerance range. So, they may be economical and can be greener substitutes for the present harsh chemical-based wastewater effluent treatment methods.

Zwitterionic surface modification is a promising strategy for nanomedicines to achieve prolonged circulation time and thus effective tumor accumulation. However, zwitterion modified nanoparticles suffer from reduced cellular internalization efficiency. Methods: A polyprodrug-based nanomedicine with zwitterionic-to-cationic charge conversion ability (denoted as ZTC-NMs) was developed for enhanced chemotherapeutic drug delivery. The polyprodrug consists of pH-responsive poly(carboxybetaine)-like zwitterionic segment and glutathione-responsive camptothecin prodrug segment. Results: The ZTC-NMs combine the advantages of zwitterionic surface and polyprodrug. Compared with conventional zwitterionic surface, the ZTC-NMs can respond to tumor microenvironment and realize ZTC surface charge conversion, thus improve cellular internalization efficiency of the nanomedicines. Conclusions: This ZTC method offers a strategy to promote the drug delivery efficiency and therapeutic efficacy, which is promising for the development of cancer nanomedicines.

Ions are ubiquitous in nature. They play a key role for many biological processes on the molecular scale, from molecular interactions, to mechanical properties, to folding, to self‐organisation and assembly, to reaction equilibria, to signalling, to energy and material transport, to recognition etc. Going beyond monovalent ions to multivalent ions, the effects of the ions are frequently not only stronger (due to the obviously higher charge), but qualitatively different. A typical example is the process of binding of multivalent ions, such as Ca2+, to a macromolecule and the consequences of this ion binding such as compaction, collapse, potential charge inversion and precipitation of the macromolecule. Here we review these effects and phenomena induced by multivalent ions for biological (macro)molecules, from the “atomistic/molecular” local picture of (potentially specific) interactions to the more global picture of phase behaviour including, e. g., crystallisation, phase separation, oligomerisation etc. Rather than attempting an encyclopedic list of systems, we rather aim for an embracing discussion using typical case studies. We try to cover predominantly three main classes: proteins, nucleic acids, and amphiphilic molecules including interface effects. We do not cover in detail, but make some comparisons to, ion channels, colloidal systems, and synthetic polymers. While there are obvious differences in the behaviour of, and the relevance of multivalent ions for, the three main classes of systems, we also point out analogies. Our attempt of a comprehensive discussion is guided by the idea that there are not only important differences and specific phenomena with regard to the effects of multivalent ions on the main systems, but also important similarities. We hope to bridge physico‐chemical mechanisms, concepts of soft matter, and biological observations and connect the different communities further., Ions play an important role for a large variety of biological processes. Compared to monovalent ions, the effects of multivalent ions are often qualitatively different and can include complex phenomena such as charge inversion and precipitation of macromolecules. In this article, we review the effects of multivalent ions on biological (macro)molecules from the molecular to the global level. Our hope is to bridge physico‐chemical, soft matter and biological aspects, thereby stimulating discussions between these scientific communities.

Cerium‐tungstate polyoxoanions are combined with caffeinium cations to give a hybrid organic‐inorganic material with anti‐HIV activity. Biochemical assays highlight the low cytotoxicity of the compounds together with their promising antiviral activity. Initial mechanistic studies provide information on their mode of action. Polyoxometalates (POMs), molecular metal oxide anions, are inorganic clusters with promising antiviral activity. Herein we report increased anti‐HIV‐1 activity of a POM when electrostatically combined with organic counter‐cations. To this end, Keggin‐type cerium tungstate POMs have been combined with organic methyl‐caffeinium (Caf) cations, and their cytotoxicity, antiviral activity and mode of action have been studied. The novel compound, Caf4K[β2‐CeSiW11O39]×H2O, exhibits sub‐nanomolar antiviral activity and inhibits HIV‐1 infectivity by acting on an early step of the viral infection cycle. This work demonstrates that combination of POM anions and organic bioactive cations can be a powerful new strategy to increase antiviral activity of these inorganic compounds., publishedVersion

Two different chromophores, namely a dipolar and an octupolar system, were prepared and their linear and nonlinear optical properties as well as their bioimaging capabilities were compared. Both contain triphenylamine as the donor and a triarylborane as the acceptor, the latter modified with cationic trimethylammonio groups to provide solubility in aqueous media. The octupolar system exhibits a much higher two‐photon brightness, and also better cell viability and enhanced selectivity for lysosomes compared with the dipolar chromophore. Furthermore, both dyes were applied in two‐photon excited fluorescence (TPEF) live‐cell imaging., Dipole versus octupole: Branching of a cationic triarylborane chromophore leads to a cooperative increase in the two‐photon absorption cross‐section and brightness, as well as higher selectivity towards lysosomes and better cell viability. The octupolar dye was demonstrated to be an ideal candidate for two‐photon excited fluorescence imaging of lysosomes in live cells.

Au2 + is a simple but crucial model system for understanding the diverse catalytic activity of gold. While the Au2 + ground state (X2Σg +) is understood reasonably well from mass spectrometry and computations, no spectroscopic information is available for its first excited state (A2Σu +). Herein, we present the vibrationally resolved electronic spectrum of this state for cold Ar‐tagged Au2 + cations. This exceptionally low‐lying and well isolated A2Σ(u) +←X2Σ(g) + transition occurs in the near‐infrared range. The observed band origin (5738 cm−1, 1742.9 nm, 0.711 eV) and harmonic Au−Au and Au−Ar stretch frequencies (201 and 133 cm−1) agree surprisingly well with those predicted by standard time‐dependent density functional theory calculations. The linearly bonded Ar tag has little impact on either the geometric or electronic structure of Au2 +, because the Au2 +⋅⋅⋅Ar bond (∼0.4 eV) is much weaker than the Au−Au bond (∼2 eV). As a result of 6 s←5d excitation of an electron from the antibonding σu * orbital (HOMO‐1) into the bonding σg orbital (SOMO), the Au−Au bond contracts substantially (by 0.1 Å)., The Au2 + cation: The high‐resolution near‐infrared spectrum of the lowest excited electronic state of this simple diatomic cation recorded by Ar‐tagging photodissociation exhibits a surprisingly simple vibronic structure. Analysis by standard TD‐DFT calculations provides the first spectroscopic information about the chemical bond in Au2 + in both the ground and excited electronic states.

In accordance with the opinion of the World Health Organization and the World Water Council the development of effective technologies for the treatment of wastewater from heavy metals for their discharge into water bodies or reuse is an urgent task nowadays. Phytoremediation biotechnologies is the most environmentally friendly and cheapest way of the treatment of wastewater, suitable for sustainable development principals. The main disadvantage of the phytoremediation is the slow speed of the process. A method for accelerating the process of phytoremediation by the combined effect of magnetic and weak electric fields is proposed. The purpose of this study is to determine the values of the parameters of the magnetic and weak electric fields that are most suitable for extracting cuprum ions from wastewater using the higher aqua plants (Lemna minor). A corresponding technological process based on the results of the study is proposed. The results have shown that the removal of copper cations from sulfate solutions effectively occurs in the initial period of time (1–5 hours) under the influence of a magnetic field with an intensity of H = 2 kA/m. Under the combined influence of an electrical current with density j = 240 μA/cm2 and a magnetic field (H = 2 kA/m) the highest rate of copper extraction by duckweed leaves is achieved. Under these conditions, the greatest growth and development of plant leaves occurs. The paper presents the results of determining of the parameters of the electrochemical release from the eluate of the spent phytomass of duckweed. It has been determined that the release of metal occurs at E = 0.32 V. An original scheme for wastewater treatment from copper with subsequent separation of copper from the spent phytomass of duckweed is proposed. In general, the presented results are a scientific justification of wastewater treatment technologies and a contribution to resolving the crisis in the field of fresh water supply. An important contribution in the circular economy is a technology recommendation proposed for recovering copper from duckweed after wastewater treatment.

Bis‐NHC stabilized germyliumylidenes [RGe(NHC)2]+ are typically Lewis basic (LB) in nature, owing to their lone pair and coordination of two NHCs to the vacant p‐orbitals of the germanium center. However, they can also show Lewis acidity (LA) via Ge−CNHC σ* orbital. Utilizing this unique electronic feature, we report the first example of bis‐NHC‐stabilized germyliumylidene [MesTerGe(NHC)2]Cl (1), (MesTer=2,6‐(2,4,6‐Me3C6H2)2C6H3; NHC= IMe4=1,3,4,5‐tetramethylimidazol‐2‐ylidene) catalyzed reduction of CO2 with amines and arylsilane, which proceeds via its Lewis basic nature. In contrast, the Lewis acid nature of 1 is utilized in the catalyzed hydroboration and cyanosilylation of carbonyls, thus highlighting the versatile ambiphilic nature of bis‐NHC stabilized germyliumylidenes., The versatile ambiphilic catalytic application of a bis‐NHC‐stabilized germyliumylidene enables a range of reductive functionalization reactions, with a particular emphasis on C=O reduction that is, CO2 and carbonyls.

A nutritional survey was conducted to study the influence of variety and rootstock on interactionamong K+, Ca++, Mg++and Na+ ions in grape during 2012-14. Soil cation contents did not correlatewith their respective contents in petioles indicating a strong antagonism among them. Quadraticrelationship of soil cations with the absorption (ratio of petiole content to soil content) of otherions revealed that the antagonism among cations was observed in case of soil K+ with Ca++ andNa+ absorption on 110R and Dog Ridge rootstocks, soil Ca+ with K+ and Mg++ and Na+ in Sonakavariety and Na+ in own rooted vines, soil Mg++ with Ca++ and Na+ also in own rooted vines; andNa++ with Ca++ and Mg++ respectively in 2A clone and Dog Ridge. Contrarily, increased absorptionof K+ by soil Ca++ on 110R, Na+ and K+ by soil Mg++ respectively in Sonaka and 110R, and Ca++by soil Na+ on Dog Ridge was also observed. All the soil cations together influenced K+ absorptionmost in Sonaka followed by Mg++ absorption in 2A clone, but Ca++ absorption on Dog Ridgefollowed by K+ on 110R.

The discharge of inadequately treated or untreated industrial wastewaters has greatly contributed to the release of contaminants into the environment, including toxic metals. Toxic metals are persistent and bioaccumulative, being their removal from wastewaters prior to release into water bodies of great concern. Literature reports the use of brown marine macroalgae for toxic metals removal from aqueous solutions as an economic and eco-friendly technique, even when applied to diluted solutions. Minor attention has been given to the application of this technique in the treatment of real wastewaters, which present a complex composition that can compromise the biosorption performance. Therefore, the main goal of this comprehensive review is to critically outline studies that: (i) applied brown marine macroalgae as natural cation exchanger for toxic metals removal from real and complex matrices; (ii) optimised the biosorption process in a fixed-bed column, which was further scaled-up to pilot plants. An overview of toxic metals sources, chemistry and toxicity, which are relevant aspects to understand and develop treatment techniques, is initially presented. The problem of water resources pollution by toxic metals and more specifically the participation of metal finishing industries in the environmental contamination are issues also covered. The current and potential decontamination methods are presented including a discussion of their advantages and drawbacks. The literature on biosorption was reviewed in detail, considering especially the ion exchange properties of cell wall constituents, such as alginate and fucoidan, and their role in metal sequestration. Besides that, a detailed description of biosorption process design, especially in continuous mode, and the application of mechanistic models is addressed.

Mycobacterium tuberculosis virulence is highly metal‐dependent with metal availability modulating the shift from the dormant to active states of M. tuberculosis infection. Rv0045c from M. tuberculosis is a proposed metabolic serine hydrolase whose folded stability is dependent on divalent metal concentration. Herein, we measured the divalent metal inhibition profile of the enzymatic activity of Rv0045c and found specific divalent transition metal cations (Cu(2+) ≥ Zn(2+) > Ni(2+) > Co(2+)) strongly inhibited its enzymatic activity. The metal cations bind allosterically, largely affecting values for k (cat) rather than K (M). Removal of the artificial N‐terminal 6xHis‐tag did not change the metal‐dependent inhibition, indicating that the allosteric inhibition site is native to Rv0045c. To isolate the site of this allosteric regulation in Rv0045c, the structures of Rv0045c were determined at 1.8 Å and 2.0 Å resolution in the presence and absence of Zn(2+) with each structure containing a previously unresolved dynamic loop spanning the binding pocket. Through the combination of structural analysis with and without zinc and targeted mutagenesis, this metal‐dependent inhibition was traced to multiple chelating residues (H202A/E204A) on a flexible loop, suggesting dynamic allosteric regulation of Rv0045c by divalent metals. Although serine hydrolases like Rv0045c are a large and diverse enzyme superfamily, this is the first structural confirmation of allosteric regulation of their enzymatic activity by divalent metals.

Here we demonstrate that binuclear dinitrosyl iron complexes with thiol-containing ligands (glutathione and mercaptosuccinate, B-DNIC-GSH and B-DNIC-MS, respectively) exert cytotoxic effects on MCF7 human breast cancer cells. We showed that they are mediated by nitrosonium cations released from these complexes (NO+). This finding is supported by the cytotoxic effect of both B-DNICs on MCF7 cells evidenced to retain or was even promoted in the presence of N-Methyl-D-glucamine dithiocarbamate (MGD). MGD recruits an iron nitrosyl group [Fe(NO)] from the iron-dinitrosyl fragment [Fe(NO)2] of B-DNIC-MS forming stable mononitrosyl complexes of iron with MGD and releasing NO+ cations from a [Fe(NO)2] fragment.

Acta crystallographica / B 77(3), 331-338 (2021). doi:10.1107/S2052520621002663, The X-ray crystal structure of the d(CGCGCG)$_2$/putrescine(2+)/K$^+$ com��plex has been determined at 0.60 �� resolution. Stereochemical restraints were used only for the putrescinium dication, and 23 bonds and 18 angles of the Z-DNA nucleotides with dual conformation. The N atoms of the putrescine(2+) dication form three direct hydrogen bonds with the N7_G atoms of three different Z-DNA molecules, plus three water-mediated hydrogen bonds with cytosine, guanine and phosphate acceptors. A unique potassium cation was also unambiguously identified in the structure, albeit at a ���0.5 occupation site shared with a water molecule, providing the first example of such a com��plex with Z-DNA. The K$^+$ cation has coordination number of eight and an irregular coordination sphere, formed by four water molecules and four O atoms from two phosphate groups of the Z-DNA, including ligands present at fractional occupancy. The structural disorder of the Z-DNA duplex is manifested by the presence of alternate conformations along the DNA backbone. Comparison of the position and interactions of putrescine(2+) in the present structure with other ultra-high-resolution structures of Z-DNA in com��plexes with Mn$^{2+}$ and Zn$^{2+}$ ions shows that the dicationic putrescinium moiety can effectively substitute these metal ions for stabilization of Z-type DNA duplexes. Furthermore, this comparison also suggests that the spermine(4+) tetracation has a higher affinity for Z-DNA than K$^+$., Published by Wiley-Blackwell, Oxford [u.a.]

Key points We report a novel method for the transient expression of SARS‐CoV‐2 envelope (E) protein in intracellular organelles and the plasma membrane of mammalian cells and Xenopus oocytes.Intracellular expression of SARS‐CoV‐2 E protein increases intra‐Golgi pH.By targeting the SARS‐CoV‐2 E protein to the plasma membrane, we show that it forms a cation channel, viroporin, that is modulated by changes of pH.This method for studying the activity of viroporins may facilitate screening for new antiviral drugs to identify novel treatments for COVID‐19. Abstract The envelope (E) protein of coronaviruses such as SARS‐CoV‐1 is proposed to form an ion channel or viroporin that participates in viral propagation and pathogenesis. Here we developed a technique to study the E protein of SARS‐CoV‐2 in mammalian cells by directed targeting using a carboxyl‐terminal fluorescent protein tag, mKate2. The wild‐type SARS‐CoV‐2 E protein can be trafficked to intracellular organelles, notably the endoplasmic reticulum–Golgi intermediate complex, where its expression increases pH inside the organelle. We also succeeded in targeting SARS‐CoV‐2 E to the plasma membrane, which enabled biophysical analysis using whole‐cell patch clamp recording in a mammalian cell line, HEK 293 cells, and two‐electrode voltage clamp electrophysiology in Xenopus oocytes. The results suggest that the E protein forms an ion channel that is permeable to monovalent cations such as Na+, Cs+ and K+. The E current is nearly time‐ and voltage‐independent when E protein is expressed in mammalian cells, and is modulated by changes of pH. At pH 6.0 and 7.4, the E protein current is activated, whereas at pH 8.0 and 9.0, the amplitude of E protein current is reduced, and in oocytes the inward E current fades at pH 9 in a time‐ and voltage‐dependent manner. Using this directed targeting method and electrophysiological recordings, potential inhibitors of the E protein can be screened and subsequently investigated for antiviral activity against SARS‐CoV‐2 in vitro and possible efficacy in treating COVID‐19., Key points We report a novel method for the transient expression of SARS‐CoV‐2 envelope (E) protein in intracellular organelles and the plasma membrane of mammalian cells and Xenopus oocytes.Intracellular expression of SARS‐CoV‐2 E protein increases intra‐Golgi pH.By targeting the SARS‐CoV‐2 E protein to the plasma membrane, we show that it forms a cation channel, viroporin, that is modulated by changes of pH.This method for studying the activity of viroporins may facilitate screening for new antiviral drugs to identify novel treatments for COVID‐19.

How to efficiently improve waste activated sludge (WAS) dewaterability is a common challenge in WAS treatment and management throughout world. The interaction energy of sludge flocs is of great importance for sludge dewaterability. In this study, the relationship among the repulsive force of sludge flocs, hydrophilic/hydrophobic characteristics of sludge flocs, and sludge dewaterability have been quantitatively and qualitatively investigated based on extended Derjaguin-Landau-Verwey-Overbeek theory for the first time. The energy barrier of sludge flocs has good correlations with sludge dewaterability (p 0.05). Trivalent cations (Al

The development of delivery vehicles for small interfering RNAs (siRNAs) remains a bottleneck to widespread clinical use. Cationic polymers represent an important class of potential delivery vehicles. In this study, we used alkyne-azide click chemistry to synthesize a variety of cationic poly(propargyl glycolide) backbone polymers to bind and deliver siRNAs. We demonstrated control over the binding interactions of these polymers and siRNAs by varying binding strength by more than three orders of magnitude. Binding strength was found to meet or exceed that of commercially available transfection agents. Our polymers effectively delivered siRNAs with no detectable cytotoxicity. Despite accumulation of siRNAs at levels comparable with commercial reagents, we did not observe silencing of the targeted protein. The implications of our results for future siRNA delivery vehicle design are discussed.

Rationale: Cationic nanocarriers present with well-known toxicities, including inflammatory toxicity, which limit their clinical application. How the cationic nanocarrier-induced inflammatory response is negatively regulated is unknown. Herein, we found that following a sublethal dose of cationic nanocarriers, the induced inflammatory response is characterized by early neutrophil infiltration and spontaneous resolution within 1 week. Methods: C57BL/6 mice were intravenously injected with a dosage of 1-100 mg/kg cationic DOTAP liposomes as well as other cationic materials. Cell necrosis was detected by flow cytometry. Release of mitochondrial DNA was quantified by qPCR via Taqman probes. Signal proteins were detected by Western blotting. PGE2 production in the supernatant was quantitated using an enzyme immunoassay (EIA). The infiltrated inflammatory cells were observed in WT mice, Ccr2-/- mice, Sting-/- mice and Tlr9-/- mice. Results: The early stage (24-48 h) inflammatory neutrophil infiltration was followed by an increasing percentage of monocytes; and, compared with WT mice, Ccr2-/- mice presented with more severe pulmonary inflammation. A previously uncharacterized population of regulatory monocytes expressing both inflammatory and immunosuppressive cytokines was identified in this model. The alteration in monocyte phenotype was directly induced by mtDNA release from cationic nanocarrier-induced necrotic cells via a STING- or TLR9-dependent pathway. Neutrophil activation was specifically inhibited by PGE2 from Ly6C+ inflammatory monocytes, and intravenous injections of dual-phenotype monocytes beneficially modified the immune response; this inhibitory effect was abolished after treatment with indomethacin. Moreover, we provide clear evidence that mitochondrial DNA activated Ly6C+ monocytes and increased PGE2 production through TLR9- or STING-mediated MAPK-NF-κB-COX2 pathways. Conclusion: Our findings suggest that Ly6C+ monocytes and mtDNA-induced Ly6C+ monocyte PGE2 production may be part of a feedback mechanism that contributes to the resolution of cationic nanocarrier-induced inflammatory toxicity and may have important implications for understanding nanoparticle biocompatibility and designing better, safer drug delivery systems.

Structural variations of the well-known guanine quartet (G4) motif in nucleic acid structures, namely substitution of two guanine bases (G) by two adenine (A) nucleobases in mutual trans positions, are discussed and studied by density functional theory (DFT) methods. This work was initiated by three findings, namely (1) that GA mismatches are compatible with complementary pairing patterns in duplex-DNA structures and can, in principle, be extended to quartet structures, (2) that GA pairs can come in several variations, including with a N1 protonated adeninium moiety (AH), and (3) that cross-linking of the major donor sites of purine nucleobases (N1 and N7) by transition metal ions of linear coordination geometries produces planar purine quartets, as demonstrated by some of us in the past. Here, possible structures of mixed AGAG quartets both in the presence of protons and alkali metal ions are discussed, and in particular, the existence of a putative four-purine, two-metal motif. Electronic supplementary material The online version of this article (10.1007/s00775-017-1507-7) contains supplementary material, which is available to authorized users.

24 páginas Contiene 4 figuras y 5 tablas. 23 referencias en las páginas 441-442 Soil formation has been studied in relation with time in a 5000-year old chronosequence on volcaniclastic beach ridges of the perhumid tropical Atlantic coast of Costa Rica. All soils are under tropical rainforest. Drainage conditions change by subsidence from excessively drained in the two youngest soils to imperfectly drained in the two oldest soils. Parent material is rather homogeneous andesitic sand with a volcanic glass component of less than 10%. It has been found that under these conditions Andisols form within 2000 years. Imperfect drainage caused mottling and accumulation of iron-coatings, as well as the formation of a thin O-horizon in the oldest profiles. Sand content of the soils decreases regularly with soil age, while the amount of fine material increases concurrently. The increase in fine material and the accumulation of organic matter cause an increase of CEC and andic properties, and a decrease in bulk density and pH with soil age. Depth of biological influence increases with soil age, but soil faunal activity is hampered in the oldest three profiles, probably by imperfect drainage. Due to the extreme leaching conditions, the sum of exchangeable cations is less than 2 cmol + kg- 1 in the B-horizons of the older soils, notwithstanding the presence of a considerable amount of weatherable primary minerals.

Conjugated polyelectrolytes (CPEs) have been widely used as reporters in colorimetric assays targeting nucleic acids. CPEs provide naked eye detection possibility by their superior optical properties however, as concentration of target analytes decrease, trace amounts of nucleic acid typically yield colorimetric responses that are not readily perceivable by naked eye. Herein, we report a pixelated analysis approach for correlating colorimetric responses of CPE with nucleic acid concentrations down to 1 nM, in plasma samples, utilizing a smart phone with an algorithm that can perform analytical testing and data processing. The detection strategy employed relies on conformational transitions between single stranded nucleic acid-cationic CPE duplexes and double stranded nucleic acid-CPE triplexes that yield distinct colorimetric responses for enabling naked eye detection of nucleic acids. Cationic poly[N,N,N-triethyl-3-((4-methylthiophen-3-yl)oxy)propan-1-aminium bromide] is utilized as the CPE reporter deposited on a polyvinylidene fluoride (PVDF) membrane for nucleic acid assay. A smart phone application is developed to capture and digitize the colorimetric response of the individual pixels of the digital images of CPE on the PVDF membrane, followed by an analysis using the algorithm. The proposed pixelated approach enables precise quantification of nucleic acid assay concentrations, thereby eliminating the margin of error involved in conventional methodologies adopted for interpretation of colorimetric responses, for instance, RGB analysis. The obtained results illustrate that a ubiquitous smart phone could be utilized for point of care colorimetric nucleic acids assays in complex matrices without requiring sophisticated software or instrumentation., Graphical abstract Image 1, Highlights • A cost effective sensory cartridge and an algorithm compatible with smart phone are developed. • Conjugated polyelectrolyte impregnated paper for detection of nucleic acid. • A pixelated assay to visualize nucleic acid down to concentration of 1 nM.

// Shuai Zhen 1, 2 , Yoichiro Takahashi 3 , Shunichi Narita 3 , Yi-Chen Yang 4 , Xu Li 1, 2 1 Center for Translational Medicine, The First Affiliated Hospital of Xi’an Jiaotong University, P.R. China 2 Key Laboratory for Tumor Precision Medicine of Shaanxi Province, The First Affiliated Hospital of Xi’an Jiaotong University, P.R. China 3 Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto, Japan 4 Tsinghua University, School of Life Sciences, Beijing, China Correspondence to: Xu Li, email: lixu56@mail.xjtu.edu.cn Keywords: CRISPR/Cas9 delivery, aptamer-liposome chimera, prostate cancer, prostate-specific membrane antigen (PSMA), polo-like kinase 1 (PLK1) Received: September 01, 2016 Accepted: December 06, 2016 Published: December 21, 2016 ABSTRACT The potent ability of CRISPR/Cas9 system to inhibit the expression of targeted gene is being exploited as a new class of therapeutics for a variety of diseases. However, the efficient and safe delivery of CRISPR/Cas9 into specific cell populations is still the principal challenge in the clinical development of CRISPR/Cas9 therapeutics. In this study, a flexible aptamer-liposome-CRISPR/Cas9 chimera was designed to combine efficient delivery and increased flexibility. Our chimera incorporated an RNA aptamer that specifically binds prostate cancer cells expressing the prostate-specific membrane antigen as a ligand. Cationic liposomes were linked to aptamers by the post-insertion method and were used to deliver therapeutic CRISPR/Cas9 that target the survival gene, polo-like kinase 1, in tumor cells. We demonstrate that the aptamer-liposome-CRISPR/Cas9 chimeras had a significant cell-type binding specificity and a remarkable gene silencing effect in vitro . Furthermore, silencing promoted a conspicuous regression of prostate cancer in vivo . Importantly, the approach described here provides a universal means of cell type–specific CRISPR/Cas9 delivery, which is a critical goal for the widespread therapeutic applicability of CRISPR/Cas9 or other nucleic acid drugs.

Sonogenetics is a promising approach for in vivo neuromodulation using ultrasound (US) to non-invasively stimulate cells in deep tissue. However, sonogenetics requires accurate transduction of US-responsive proteins into target cells. Here, we introduce a non-invasive and non-viral approach for intracerebral gene delivery. This approach utilizes temporary ultrasonic disruption of the blood-brain barrier (BBB) to transfect neurons at specific sites in the brain via DNA that encodes engineered US-responsive protein (murine Prestin (N7T, N308S))-loaded microbubbles (pPrestin-MBs). Prestin is a transmembrane protein that exists in the mammalian auditory system and functions as an electromechanical transducer. We further improved the US sensitivity of Prestin by introducing specific amino acid substitutions that frequently occur in sonar species into the mouse Prestin protein. We demonstrated this concept in mice using US with pPrestin-MBs to non-invasively modify and activate neurons within the brain for spatiotemporal neuromodulation. Method: MBs composed of cationic phospholipid and C3F8 loaded with mouse Prestin plasmid (pPrestin) via electrostatic interactions. The mean concentration and size of the pPrestin-MBs were (16.0 ± 0.2) × 109 MBs/mL and 1.1 ± 0.2 μm, respectively. SH-SY5Y neuron-like cells and C57BL mice were used in this study. We evaluated the gene transfection efficiency and BBB-opening region resulting from pPrestin-MBs with 1-MHz US (pressure = 0.1-0.5 MPa, cycle = 50-10000, pulse repetition frequency (PRF): 0.5-5 Hz, sonication time = 60 s) using green fluorescence protein (Venus) and Evans blue staining. Results: The maximum pPrestin expression with the highest cell viability occurred at a pressure of 0.5 MPa, cycle number of 5000, and PRF of 1 Hz. The cellular transfection rate with pPrestin-MBs and US was 20.2 ± 2.5%, which was 1.5-fold higher than that of commercial transfection agents (LT-1). In vivo data suggested that the most profound expression of pPrestin occurred at 2 days after performing pPrestin-MBs with US (0.5 MPa, 240 s sonication time). In addition, no server erythrocyte extravasations and apoptosis cells were observed at US-sonicated region. We further found that with 0.5-MHz US stimulation, cells with Prestin expression were 6-fold more likely to exhibit c-Fos staining than cells without Prestin expression. Conclusion: Successful activation of Prestin-expressing neurons suggests that this technology provides non-invasive and spatially precise selective modulation of one or multiple specific brain regions.

Reactions of 2-carbonyl- and 2-hydroxy(or methoxy)alkylbenzimidazoles with arenes in the Brønsted superacid TfOH resulted in the formation of the corresponding Friedel–Crafts reaction products, 2-diarylmethyl and 2-arylmethyl-substituted benzimidazoles, in yields up to 90%. The reaction intermediates, protonated species derived from starting benzimidazoles in TfOH, were thoroughly studied by means of NMR and DFT calculations and plausible reaction mechanisms are discussed.

The ionization of 1,1‐dihydridocyclopentasilane 7 has been found to yield the cyclic polysilanylsilyl cation 8 instead of the expected hydrogen‐substituted silylium ion 6. The silyl cation 8 is stabilized by the formation of an intramolecular Si−H−Si bridge, which also provides the thermodynamic driving force for its formation. In general, the preference for the formation of Si−H−Si bridges can be used to scavenge and identify transient intermediates in the Lewis acid induced rearrangement of polysilanes. The validity of this concept has been demonstrated for one central step in this chemistry, the ring‐contraction reaction of cyclohexasilanes to form silylcyclopentasilanes.

Purpose Personalized peptide-based cancer vaccines will be composed of multiple patient specific synthetic long peptides (SLPs) which may have various physicochemical properties. To formulate such SLPs, a flexible vaccine delivery system is required. We studied whether cationic liposomes are suitable for this purpose. Methods Fifteen SIINFEKL T cell epitope-containing SLPs, widely differing in hydrophobicity and isoelectric point, were separately loaded in cationic liposomes via the dehydration-rehydration method. Particle size and polydispersity index (PDI) were measured via dynamic light scattering (DLS), and zeta potential with laser Doppler electrophoresis. Peptide loading was fluorescently determined and the immunogenicity of the formulated peptides was assessed in co-cultures of dendritic cells (DCs) and CD8+ T-cells in vitro. Results All SLPs were loaded in cationic liposomes by using three different loading method variants, depending on the SLP characteristics. The fifteen liposomal formulations had a comparable size (< 200 nm), PDI (< 0.3) and zeta potential (22–30 mV). Cationic liposomes efficiently delivered the SLPs to DCs that subsequently activated SIINFEKL-specific CD8+ T-cells, indicating improved immunological activity of the SLPs. Conclusion Cationic liposomes can accommodate a wide range of different SLPs and are therefore a potential delivery platform for personalized cancer vaccines. Electronic supplementary material The online version of this article (10.1007/s11095-018-2490-6) contains supplementary material, which is available to authorized users.

This paper proposes a green environment-friendly Bacillus subtilis to mineralize and consolidate waste incineration fly ash and heavy metal cations, and there is no harmful by-product in the mineralization process. Different phosphate products can be prepared, and are more stable than the microbially-induced carbonate precipitation (MICP) in nature. Typical heavy metal oxides were mainly PbO, ZnO, CdO, NiO, CuO and Cr2O3 in the chemical composition of waste incineration fly ash. Microstructure and chemical composition of waste incineration fly ash before and after treatment were characterized by powder X-ray diffraction (XRD) analysis and scanning electron microscopy. Scanning electron microscopy (SEM) images showed that the morphology of the Bacillus subtilis was mainly a rod-like structure. The optimal hydrolysis dosage of the organic phosphate monoester sodium salt was 0.2mol in the bacterial solution (1L, 20 g/L). The optimum required mass of the bacterial powder was 15 g/kg in treatment process of the waste incineration fly ash. The initial concentration of lead ions was 40.28 mg/L in waste incineration fly ash solution. After the optimum dosage treatment, the removal efficiency of lead ions was 78.15%, 79.64%, 77.70% and 80.14% when curing time was 1, 2, 4 and 6d, respectively. The waste incineration fly ash had a Shore hardness of 22 after the optimum amount of bacterial liquid treatment. Results of wind erosion test showed that the wind erosion rate of waste incineration fly ash was 2.6, 0, 0, 0, 0 and 0 g/h when blank group, deionized water, 100, 200, 300 and 400 mL of bacterial solutions treated, respectively. The bio-mineralization method provides an approach for the safe disposal of heavy metals in the contaminated areas of tailings, electroplating sewage, waste incineration plants, and so on.

DNA quadruplexes have been the subject of investigation because of their biological relevance and because of their potential application in supramolecular chemistry. Similarly, RNA quadruplexes are now gaining increasing attention. Although DNA and RNA quadruplexes are structurally very similar, the latter show higher stability. In this study we report dispersion‐corrected density functional theory (DFT‐D) quantum chemical calculations that were undertaken to understand the difference in stabilities of RNA and DNA quadruplexes. The smallest meaningful model of a stack of quartets, interacting with alkali metal cations, was simulated in an aqueous environment. The energy decomposition analysis allows for in‐depth examination of the interaction energies, emphasising the role of noncovalent interactions and better electrostatics in determining RNA‐GQs higher stabilities, particularly pinpointing the role of the extra 2′‐OH groups. Furthermore, our computations present new insights on why the cation is required for self‐assembly: unexpectedly the cation is not necessary to relieve the repulsion between the oxygen atoms in the central cavity, but it is needed to overcome the entropic penalty.

Non‐natural oligonucleotides represent important (bio)chemical tools and potential therapeutic agents. Backbone modifications altering hybridization properties and biostability can provide useful analogues. Here, we employ an artificial nucleosyl amino acid (NAA) motif for the synthesis of oligonucleotides containing a backbone decorated with primary amines. An oligo‐T sequence of this cationic DNA analogue shows significantly increased affinity for complementary DNA. Notably, hybridization with DNA is still governed by Watson–Crick base pairing. However, single base pair mismatches are tolerated and some degree of sequence‐independent interactions between the cationic NAA backbone and fully mismatched DNA are observed. These findings demonstrate that a high density of positive charges directly connected to the oligonucleotide backbone can affect Watson–Crick base pairing. This provides a paradigm for the design of therapeutic oligonucleotides with altered backbone charge patterns.

δ‐Cadinene synthase is a sesquiterpene cyclase that utilises the universal achiral precursor farnesyl diphosphate (FDP) to generate predominantly the bicyclic sesquiterpene δ‐cadinene and about 2 % germacradien‐4‐ol, which is also generated from FDP by the cyclase germacradien‐4‐ol synthase. Herein, the mechanism by which sesquiterpene synthases discriminate between deprotonation and reaction with a nucleophilic water molecule was investigated by site‐directed mutagenesis of δ‐cadinene synthase. If W279 in δ‐cadinene synthase was replaced with various smaller amino acids, the ratio of alcohol versus hydrocarbon product was directly proportional to the van der Waals volume of the amino acid side chain. DCS‐W279A is a catalytically highly efficient germacradien‐4‐ol synthase (k cat/K M=1.4×10−3 μm s−1) that produces predominantly germacradien‐4‐ol in addition to 11 % δ‐cadinene. Water capture is not achieved through strategic positioning of a water molecule in the active site, but through a coordinated series of loop movements that allow bulk water access to the final carbocation in the active site prior to product release.

The remediation of a real textile wastewater aiming its reuse in the textile industry was carried out by integrating two processes: (i) a chemical or electrochemical advanced oxidation process (AOP or EAOP) based on Fenton's reaction for organics degradation, and (ii) a cation exchange process using marine macroalgae for removal of the iron acting in the Fenton's reaction based processes. Four AOPs/EAOPs at acidic pH 2.8 were tested: Fenton, photo-Fenton with ultraviolet A (UVA) radiation (PF/UVA), electro-Fenton (EF) and photoelectro-Fenton with UVA radiation (PEF/UVA). These processes provided very high color removals. After a running time of 45 min, the color removals were 68–95% for the Fenton process, 76–94% for the EF process, 80–98% for the PF/UVA process and 85–100% for the PEF/UVA process. In contrast, the mineralization was negligible for all the processes, indicating the generation/presence of persistent colorless compounds. The PF process was selected as first treatment stage due to its ability for color removal and related lower costs. A set of six marine macroalgae (Gracilaria caudata, Gracilaria cervicornis, Ascophyllum nodosum, Fucus spiralis, Laminaria hyperborea and Pelvetia canaliculata) were tested for iron uptake. Laminaria hyperborea showed the highest ion exchange capacity and affinity for iron species. Its application allowed the removal of all the iron acting in the PF process (3.4 mg/L). The textile wastewater resulting from the application of PF process followed by cation exchange with Laminaria hyperborea was successfully reused in scouring, bleaching and dyeing processes.

Natural-based polyelectrolytes (PELs), with all the advantages coming from being produced from renewable and biodegradable sources, are a potential solution for the removal of dyes from wastewater. In this work, surplus Eucalyptus bleached cellulose fibres from a paper mill were modified to increase the charge and solubility of cellulose. First, reactive aldehyde groups were introduced in the cellulose backbone by periodate oxidation of cellulose. Further modification with alkylammonium produced positively charged cellulose-based PELs. The final products were characterized by several analytical techniques. The PEL with the highest substitution degree of cationic groups was evaluated for its performance in decolouration processes, bentonite being used as aid. This was found to be effective for colour removal of either anionic or cationic dyes. Bio-PELs can thus be considered as very favourable eco-friendly flocculation agents for decolouration of harsh effluents from several industries, considering their biodegradable nature and thus the ability to produce less sludge.

Biological properties of ionic liquids (ILs) have been usually tested with the help of standard biodegradation or ecotoxicity tests. So far, several articles on the identification of intermediate metabolites of microbiological decay of ILs have been published. Simultaneously, the number of novel ILs with unrecognized characteristics regarding biodegradability and effect on organisms and environment is still increasing. In this work, seven imidazolium ionic liquids of different chemical structure were studied. Three of them are 1-alkyl-3-methyl-imidazolium bromides, while the other four are tetra- or completely substituted imidazolium iodides. This study focused on the identification of intermediate metabolites of the aforementioned ionic liquids subjected to biodegradation in a laboratory activated sludge system. Both fully substituted ionic liquids and 1-ethyl-3-methyl-imidazolium bromide were barely biodegradable. In the case of two of them, no biotransformation products were detected. The elongation of the alkyl side chain made the IL more susceptible for microbiological decomposition. 1-Decyl-3-methyl-imidazolium bromide was biotransformed most easily. Its primary biodegradation up to 100 % could be achieved. Nevertheless, the cleavage of the imidazolium ring has not been observed. Electronic supplementary material The online version of this article (doi:10.1007/s10532-015-9747-0) contains supplementary material, which is available to authorized users.

The use of sewage sludge in agriculture brings the risk of microbiological and parasitological contamination of soil, ground and surface water, as well as cultivated plants. Therefore prior the application to the soil, sewage sludge must be examined, among others, for the presence of live eggs of intestinal parasites. However, the efficiency of commonly used for this purpose parasitological methods is not satisfactory. This is probably due to the presence of flocculants in the sediments used in the dehydration process. The objective of the study was analysis of the effect of flocculant (cationic acrylamide) on the possibilities of isolation of parasite eggs from dehydrated sewage sludge. For this purpose 10 samples of sewage sludge were prepared: 5 containing flocculant and 5 without flocculant. Samples were tested by flotation method according to Quinn. From sewage sludge free of flocculant, 67.8 eggs were isolated, on average, whereas from sludge containing flocculant - only 2.8 eggs. The experiments confirmed that the isolation of eggs from sewage sludge containing flocculant (cationic acrylamide) is much more difficult than from sludge free from this substance and therefore the simple parasitological methods should not be used to examine the dehydrated sewage sludge.

In this work, fly ash (FA) and polydiallyldimethylammonium chloride (PDADMAC) were utilized to treat the spent liquor (SL) of thermomechanical pulping (TMP) process in an effort to remove its lignocelluloses. The incorporation of PDADMAC into the system reduced the dosage of FA required for achieving acceptable lignocellulose removals. The maximum lignocellulose removals of 81%, 78%, 56%, 53% and 97% were achieved for lignin, hemicellulose, COD, BOD, and turbidity via treating SL with 100 g/L of FA at 25 °C for 60 min and subsequently treating with 100 mg/L of PDADMAC at 25 °C for 30 min, respectively. Comparing the two-step processes, FA pretreatment with PDADMAC post treatment was more effective than the two step process of PDADMAC pretreatment and FA post treatment. In this case, the FA pretreatment generated metal-organic complexes, and the addition of PDADMAC facilitated the formation of large flocs that could be separated from the system readily. A one stage process of combined PDADMAC and FA was less effective than the two-stage process of FA and PDADMAC treatments in removing lignocelluloses from SL.

The electromagnetic field (EMF) is newly considered as an exogenous environmental stimulus that is closely related to ion transportation on the cellular membrane, maintaining the internal ionic homeostasis. Cation transports of Ca(2+) and other metal ions, Cd(2+), Zn(2+), and Mn(2+)were studied in terms of the external Ca(2+) stress, [Ca(2+)](ext), and exposure to the physical EMF. A specific yeast strain K667 was used for controlling CAX5 (cation/H(+) exchanger) expression. Culture samples were exposed to 60 Hz, 0.1 mT sinusoidal or square magnetics waves, and intracellular cations of each sample were measured and analyzed. AtCAX5 transformant yeast grew normally under the metallic stress. However, the growth of the control group was significantly inhibited under the same cation concentration; 60 Hz and 0.1 mT magnetic field enhanced intracellular cation concentrations significantly as exposure time increased both in the AtCAX5 transformed yeast and in the control group. However, the AtCAX5-transformed yeast showed higher concentration of the intracellular cations than the control group under the same exposure EMF. AtCAX5-transformed yeasts displayed an increment in [Ca(2+)](int), [K(+)](int), [Na(+)](int), and [Zn(2+)](int) concentration under the presence of both sinusoidal and square-waved EMF stresses compared to the control group, which shows that AtCAX5 expressed in the vacuole play an important role in maintaining the homeostasis of intracellular cations. These findings could be utilized in the cultivation of the crops which were resistant to excessive exogenous ions or in the production of biomass containing a large proportion of ions for nutritional food or in the bioremediation process in metal-polluted environments.

There is an urgent need to identify new, non‐traditional antimicrobials. The discovery of new polymeric antimicrobials is limited by current low‐throughput synthetic tools, which means that limited chemical space has been explored. Herein, we employ photochemical “in‐air” reversible addition–fragmentation chain‐transfer (RAFT) polymerization with microwell plates, using liquid‐handling robots to assemble large libraries of cationic polymers, without the need for degassing or purification steps, facilitating transfer to screening. Several lead polymers were identified including a co‐polymer with propylene glycol side chains with significantly enhanced antimicrobial activity and increased therapeutic window. Mechanistic studies showed that this polymer was bacteriostatic, and surprisingly did not lyse the cell membranes, implying an alternative mode of action. This versatile method using simple robotics will help to develop new biomaterials with emergent properties.

We report on the influence of pH and monovalent/divalent cations on the catalytic current response, internal electron transfer (IET), and structure of fructose dehydrogenase (FDH) by using amperometry, spectrophotometry, and circular dichroism (CD). Amperometric measurements were performed on graphite electrodes, onto which FDH was adsorbed and the effect on the response current to fructose was investigated when varying the pH and the concentrations of divalent/monovalent cations in the contacting buffer. In the presence of 10 mM CaCl2, a current increase of up to ≈ 240% was observed, probably due to an intra-complexation reaction between Ca2+ and the aspartate/glutamate residues found at the interface between the dehydrogenase domain and the cytochrome domain of FDH. Contrary to CaCl2, addition of MgCl2 did not show any particular influence, whereas addition of monovalent cations (Na+ or K+) led to a slight linear increase in the maximum response current. To complement the amperometric investigations, spectrophotometric assays were carried out under homogeneous conditions in the presence of a 1-electron non-proton-acceptor, cytochrome c, or a 2-electron-proton acceptor, 2,6-dichloroindophenol (DCIP), respectively. In the case of cytochrome c, it was possible to observe a remarkable increase in the absorbance up to 200% when 10 mM CaCl2 was added. However, by further increasing the concentration of CaCl2 up to 50 mM and 100 mM, a decrease in the absorbance with a slight inhibition effect was observed for the highest CaCl2 concentration. Addition of MgCl2 or of the monovalent cations shows, surprisingly, no effect on the electron transfer to the electron acceptor. Contrary to the case of cytochrome c, with DCIP none of the cations tested seem to affect the rate of catalysis. In order to correlate the results obtained by amperometric and spectrophotometric measurements, CD experiments have been performed showing a great structural change of FDH when increasing the concentration CaCl2 up to 50 mM, at which the enzyme molecules start to agglomerate, hindering the substrate access to the active site probably due to a chelation reaction occurring at the enzyme surface with the glutamate/aspartate residues. Graphical AbstractFructose dehydrogenase (FDH) consists of three subunits, but only two are involved in the electron transfer process: (I) 2e−/2H+ fructose oxidation, (II) internal electron transfer (IET), (III) direct electron transfer (DET) through 2 heme c; FDH activity either in solution or when immobilized onto an electrode surface is enhanced about 2.5-fold by adding 10 mM CaCl2 to the buffer solution, whereas MgCl2 had an “inhibition” effect. Moreover, the additions of KCl or NaCl led to a slight current increase Electronic supplementary material The online version of this article (10.1007/s00216-018-0991-0) contains supplementary material, which is available to authorized users.