The Formation of Imidazolium Salt Intimate (Contact) Ion Pairs in Solution

1‐n‐Butyl‐2,3‐dimethylimidazolium (BMMI) ionic liquids (ILs) associated with different anions undergo H/D exchange preferentially at 2‐Me group of the imidazolium in deuterated solvents. This process is mainly related to the existence of ion pairs rather than the anion basicity. The H/D exchange occurs in solvents (CDCl₃ and MeCN for instance) in which intimate contact ion pairs are present and the anion possesses a labile H in its structure, such as hydrogen carbonate and prolinate. In D₂O, separated ion pairs are formed and the H/D exchange does not occur. A plausible catalytic cycle is that the IL behaves as a neutral base in the course of all H/D exchange processes. NMR experiments, density functional calculations, and molecular dynamics simulations corroborate these hypotheses.     

Bioactivity and chemical composition of the essential oil from the leaves of Guatteria australis A.St.-Hil

Essential oil from the leaves of Guatteria australis was obtained by hydrodistillation, analyzed by Gas Chromatography coupled to Mass Spectromery (GC–MS) and their antiproliferative, antileishmanial, antibacterial, antifungal and antioxidant activities were also evaluated. Twenty-three compounds were identified among which germacrene B (50.66%), germacrene D (22.22%) and (E)-caryophyllene (8.99%) were the main compounds. The highest antiproliferative activity was observed against NCI-ADR/RES (TGI = 31.08 μg/ml) and HT-29 (TGI = 32.81 μg/ml) cell lines. It also showed good antileishmanial activity against Leishmania infantum (IC₅₀ = 30.71 μg/ml). On the other hand, the oil exhibited a small effect against Staphylococcus aureus ATCC 6538, S. aureus ATCC 14458 and Escherichia coli ATCC 10799 (MIC = 250 μg/ml), as well as small antioxidant activity (457 μmol TE/g) assessed through ORAC_FL assay. These results represent the first report regarding chemical composition and bioactivity of G. australis essential oil.     

Nonvisual Opsins and the Regulation of Peripheral Clocks by Light and Hormones

The molecular clock machinery is conserved throughout evolution. However, how environmental cues are perceived has evolved in such a way that peripheral clocks in mammals require a variety of signals, including hormones. On the other hand, in nonmammalian cells able to directly detect light, light seems to play a major role in the synchronization of the clock. The interaction between perception of circadian light by nonvisual opsins and hormones will be discussed under the perspective of clock synchronization at the molecular level.     

The Biochemical Mechanisms of Antimicrobial Photodynamic Therapy†

The unbridled dissemination of multidrug-resistant pathogens is a major threat to global health and urgently demands novel therapeutic alternatives. Antimicrobial photodynamic therapy (aPDT) has been developed as a promising approach to treat localized infections regardless of drug resistance profile or taxonomy. Even though this technique has been known for more than a century, discussions and speculations regarding the biochemical mechanisms of microbial inactivation have never reached a consensus on what is the primary cause of cell death. Since photochemically generated oxidants promote ubiquitous reactions with various biomolecules, researchers simply assumed that all cellular structures are equally damaged. In this study, biochemical, molecular, biological and advanced microscopy techniques were employed to investigate whether protein, membrane or DNA damage correlates better with dose-dependent microbial inactivation kinetics. We showed that although mild membrane permeabilization and late DNA damage occur, no correlation with inactivation kinetics was found. On the other hand, protein degradation was analyzed by three different methods and showed a dose-dependent trend that matches microbial inactivation kinetics. Our results provide a deeper mechanistic understanding of aPDT that can guide the scientific community toward the development of optimized photosensitizing drugs and also rationally propose synergistic combinations with antimicrobial chemotherapy.     

Immobilized bacterial luciferase and its applications

This review discusses the properties of the bioluminescent bacterial system as well as the methods for immobilization of bacterial luciferases and for their co-immobilization with other enzymes. The analytical systems using immobilized bacterial luciferases and their applications in analytical biochemistry and biotechnology have been described.     

Electrochemical Detection of Guanine‐methylation Using Glassy Carbon Electrode

This work presents a study of the electrochemical oxidation of 7‐methylguanine (7‐mGua) in aqueous solution at glassy carbon electrode by cyclic voltammetry, differential pulse voltammetry, square wave voltammetry and electrochemical impedance spectrometry. The anodic behaviour of 7‐mGua was compared with the electro‐oxidation of guanine and 7‐methylguanosine. The results demonstrated that the methyl and ribose groups are not electroactive but strongly influence the oxidation mechanism of these species. The oxidation of 7‐mGua occurred in a single pH‐dependent step, with the withdrawal of two electrons and two protons of C8, to form 8‐oxo‐7‐methylguanine, while the electro‐oxidation of 7‐methylguanosine also occurred in a single pH‐dependent step, however, with the withdrawal of one electron and one proton of C8 to form a hydroxylated product, since its oxidation to 8‐oxo‐7‐methylguanosine is hindered by the presence of the pendant groups. In addition, the oxidation of 7‐mGua was investigated in the presence of DNA and DNA‐bases, leading to the conclusion that the formation of 7‐mGua, from an interaction of DNA with an alkylating agent, would cause an increase on the deoxyguanosine peak current of the DNA‐biosensor, with no interference of any free DNA bases, which demonstrated that DNA‐electrochemical biosensors find application on detecting DNA methylation, opening a new avenue for applications of DNA biosensors.     

Catalytic DNA-based fluorescence polarization chiral sensing platform for l-histidine detection at trace level

This paper reports a novel fluorescence polarization (FP) chiral sensor approach based on a catalytic DNA. This platform involves an enzyme module (E), which was able to trigger the l-histidine-dependent cleavage of an RNA phosphoester bond of a substrate domain (S), whereas it did not accept the d-enantiomer as cofactor. Two assay formats were proposed, based on bi- and unimolecular strategies. The bimolecular design was related to the use of separate E and fluorescently labelled S* sequences. The two oligonucleotide strands were pre-assembled via complementary regions at their extremities. As the result of the large molecular volume of the formed assembly, the S* probe displayed a high fluorescence anisotropy signal. Upon addition of the l-histidine, the DNAzyme cleaved the phosphoester bond of the S* component, leading to the loss of stem stability and the release of single-stranded products of lower size. This was accompanied by a significant decrease in the fluorescence anisotropy response. As a simpler alternative, the unimolecular design, where E and S sequences are linked together through a loop to form a single fluorescent probe E-S*, was also investigated. It was found that the unimolecular approach provided an improved FP response relative to the bimolecular one. Under optimized operating conditions, such a chiral sensing platform allowed the detection of as low as 0.05 % of the l-histidine enantiomer in a non-racemic mixture.     

Antigenotoxic Effect Against Ultraviolet Radiation‐induced DNA Damage of the Essential Oils from Lippia Species

The antigenotoxicity against ultraviolet radiation (UV)‐induced DNA damage of essential oils (EO) from Lippia species was studied using SOS Chromotest. Based on the minimum concentration that significantly inhibits genotoxicity, the genoprotective potential of EO from highest to lowest was Lippia graveolens, thymol‐RC ≈ Lippia origanoides, carvacrol‐RC ≈ L. origanoides, thymol‐RC > Lippia alba, citral‐RC ≈ Lippia citriodora, citral‐RC ≈ Lippia micromera, thymol‐RC > L. alba, myrcenone‐RC. EO from L. alba, carvone/limonene‐RC, L. origanoides, α‐phellandrene‐RC and L. dulcis, trans‐β‐caryophyllene‐RC did not reduce the UV genotoxicity at any of the doses tested. A gas chromatography with flame ionization detection analysis (GC‐FID) was conducted to evaluate the solubility of the major EO constituents under our experimental conditions. GC‐FID analysis showed that, at least partially, major EO constituents were water‐soluble and therefore, they were related with the antigenotoxicity detected for EO. Constituents such as p‐cymene, geraniol, carvacrol, thymol, citral and 1,8‐cineole showed antigenotoxicity. The antioxidant activity of EO constituents was also determined using the oxygen radical antioxidant capacity (ORAC) assay. The results showed that the antigenotoxicity of the EO constituents was unconnected with their antioxidant activity. The antigenotoxicity to different constituent binary mixtures suggests that synergistic effects can occur in some of the studied EO.     

Viability of Using Glycerin as a Co-substrate in Anaerobic Digestion of Sugarcane Stillage (Vinasse): Effect of Diversified Operational Strategies

Vinasse, from sugar and ethanol production, stands out as one of the most problematic agroindustry wastes due to its high chemical oxygen demand, large production volume, and recalcitrant compounds. Therefore, the viability of using glycerin as a co-substrate in vinasse anaerobic digestion was tested, to increase process efficiency and biogas productivity. The effect of feeding strategy, influent concentration, cycle length, and temperature were assessed to optimize methane production. Glycerin (1.53% v/v) proved to be a good co-substrate since it increased the overall methane production in co-digestion assays. CH₄ productivity enhanced exponentially as influent concentration increased, but when temperature was increased to 35 °C, biogas production was impaired. The highest methane productivity and yield were achieved using fed-batch mode, at 30 °C and at an organic loading rate of 10.1 kg COD m⁻³ day⁻¹: 139.32 mol CH₄ m⁻³ day⁻¹, 13.86 mol CH₄ kg COD_applied, and 15.30 mol CH₄ kg COD_removed. Methane was predominantly produced through the hydrogenotrophic route. In order to treat all the vinasse produced by a mid-size sugar and ethanol plant, nine reactors with 7263.4 m³ each would be needed. The energy generated by burning the biogas in boilers would reach approximately 92,000 MW h per season and could save up to US$ 240,000.00 per month in diesel oil demand.