Radical in the Peroxide-Produced F-Type Ferryl Form of Bovine Cytochrome c Oxidase

The reduction of O₂ in respiratory cytochrome c oxidases (CcO) is associated with the generation of the transmembrane proton gradient by two mechanisms. In one of them, the proton pumping, two different types of the ferryl intermediates of the catalytic heme a₃-Cu_B center P and F forms, participate. Equivalent ferryl states can be also formed by the reaction of the oxidized CcO (O) with H₂O₂. Interestingly, in acidic solutions a single molecule of H₂O₂ can generate from the O an additional F-type ferryl form (F^•) that should contain, in contrast to the catalytic F intermediate, a free radical at the heme a₃-Cu_B center. In this work, the formation and the endogenous decay of both the ferryl iron of heme a₃ and the radical in F^• intermediate were examined by the combination of four experimental approaches, isothermal titration calorimetry, electron paramagnetic resonance, and electronic absorption spectroscopy together with the reduction of this form by the defined number of electrons. The results are consistent with the generation of radicals in F^• form. However, the radical at the catalytic center is more rapidly quenched than the accompanying ferryl state of heme a₃, very likely by the intrinsic oxidation of the enzyme itself.     

Auxin Metabolite Profiling in Isolated and Intact Plant Nuclei

The plant nucleus plays an irreplaceable role in cellular control and regulation by auxin (indole-3-acetic acid, IAA) mainly because canonical auxin signaling takes place here. Auxin can enter the nucleus from either the endoplasmic reticulum or cytosol. Therefore, new information about the auxin metabolome (auxinome) in the nucleus can illuminate our understanding of subcellular auxin homeostasis. Different methods of nuclear isolation from various plant tissues have been described previously, but information about auxin metabolite levels in nuclei is still fragmented and insufficient. Herein, we tested several published nucleus isolation protocols based on differential centrifugation or flow cytometry. The optimized sorting protocol leading to promising yield, intactness, and purity was then combined with an ultra-sensitive mass spectrometry analysis. Using this approach, we can present the first complex report on the auxinome of isolated nuclei from cell cultures of Arabidopsis and tobacco. Moreover, our results show dynamic changes in auxin homeostasis at the intranuclear level after treatment of protoplasts with free IAA, or indole as a precursor of auxin biosynthesis. Finally, we can conclude that the methodological procedure combining flow cytometry and mass spectrometry offers new horizons for the study of auxin homeostasis at the subcellular level.     

Methodology to break test for surfactant-based fracturing gel

Surfactant-based fracturing gels are considered as clean gels due to the absence of insoluble residues in their composition. This kind of fluid has been developed to minimize or eliminate damages to fractures. In view of this, new studies about properties of this kind of gel have become more and more important. Gel break is an important parameter for hydraulic fracturing gels. This research was carried out with the purpose of developing a laboratory break methodology for surfactant-based fracturing gel. It was observed that the studied gel presented optimum break results, due to a viscosity decrease to 10 mPa s in the first 6 h of the test.     

Reversible Lectin Binding to Glycan-Functionalized Graphene

The monolayer character of two-dimensional materials predestines them for application as active layers of sensors. However, their inherent high sensitivity is always accompanied by a low selectivity. Chemical functionalization of two-dimensional materials has emerged as a promising way to overcome the selectivity issues. Here, we demonstrate efficient graphene functionalization with carbohydrate ligands—chitooligomers, which bind proteins of the lectin family with high selectivity. Successful grafting of a chitooligomer library was thoroughly characterized, and glycan binding to wheat germ agglutinin was studied by a series of methods. The results demonstrate that the protein quaternary structure remains intact after binding to the functionalized graphene, and that the lectin can be liberated from the surface by the addition of a binding competitor. The chemoenzymatic assay with a horseradish peroxidase conjugate also confirmed the intact catalytic properties of the enzyme. The present approach thus paves the way towards graphene-based sensors for carbohydrate–lectin binding.     

Targeting of the Mitochondrial TET1 Protein by Pyrrolo[3,2-b]pyrrole Chelators

Targeting of epigenetic mechanisms, such as the hydroxymethylation of DNA, has been intensively studied, with respect to the treatment of many serious pathologies, including oncological disorders. Recent studies demonstrated that promising therapeutic strategies could potentially be based on the inhibition of the TET1 protein (ten-eleven translocation methylcytosine dioxygenase 1) by specific iron chelators. Therefore, in the present work, we prepared a series of pyrrolopyrrole derivatives with hydrazide (1) or hydrazone (2–6) iron-binding groups. As a result, we determined that the basic pyrrolo[3,2-b]pyrrole derivative 1 was a strong inhibitor of the TET1 protein (IC₅₀ = 1.33 μM), supported by microscale thermophoresis and molecular docking. Pyrrolo[3,2-b]pyrroles 2–6, bearing substituted 2-hydroxybenzylidene moieties, displayed no significant inhibitory activity. In addition, in vitro studies demonstrated that derivative 1 exhibits potent anticancer activity and an exclusive mitochondrial localization, confirmed by Pearson’s correlation coefficient of 0.92.     

Influence of cosurfactant in microemulsion systems for color removal from textile wastewater

Microemulsion systems have proved very efficient in color removal from textile wastewater using n‐butyl alcohol as cosurfactant. The cosurfactant has a very important role in microemulsified systems, as it is responsible for their stability, mainly in systems formed by ionic surfactants. Although very efficient, n‐butyl alcohol is partially soluble in water, which would permit its passage to the effluent. In this work, isoamyl and octyl alcohols, due to their lower solubility in water, were used as cosurfactants to evaluate their influence in color removal. The colorimetry system used was the CIE L*a*b* (CIELAB) color space and CIE L*a*b* color difference (ΔE*_ab). The wastewater used in this study was the reactive exhausted dye liquor from a dye house (first discharge) containing Procion Yellow H‐E4R (CI Reactive Yellow 84), Procion Blue H‐ERD (CI Reactive Blue 160) and Procion Red H‐E3B (CI Reactive Red 120). The obtained results were modeled using an experimental planning (the Scheffé net) and evaluated through isoresponse diagrams by correlation graphs between experimental values and those obtained by the models with an error lower than 4%. All the optimized systems were very efficient and more than 94% of the dyes contained in the effluent were removed. The microemulsion load capacity was determined using a synthetic solution containing, the same dyes present in the reactive exhausted dyebath, but 200 times concentrated, and the dyes extraction was more than 99.6%. By comparing n‐butyl, isoamyl and octyl alcohols, it was observed that the system using isoamyl alcohol presented slightly better color removal and much higher load capacity than the n‐butyl and octyl alcohols. Copyright © 2004 Society of Chemical Industry