Semisynthetic and Enzyme-Mediated Conjugate Preparations Illuminate the Ubiquitination-Dependent Aggregation of Tau Protein

In the brain of individuals with Alzheimer's disease, the regulatory protein ubiquitin is found conjugated to different lysine residues of tau protein assembled into pathological paired helical filaments. To shed light on the hitherto unexplored ubiquitination-linked conformational transitions of tau, the availability of in vitro ubiquitin conjugation methods is of primary importance. In our work, we focused on the four-repeat domain of tau and assembled an enzymatic machinery formed by UBE1, Ubc13, and CHIP enzymes. The enzymatic reaction resulted in monoubiquitination at multiple sites, reminiscent of the ubiquitination pattern observed in vivo. We further exploited chemoselective disulfide coupling reactions to construct three tau regioisomers with site-specific monoubiquitination. Protein aggregation experiments revealed that the multiple enzyme-derived products were unable to convert into amyloid fibrils, while the semisynthetic conjugates exhibited diverse capability to form filaments. This study contributes novel insight into the effects of a key post-translational modification on aberrant protein self-assembly.     

Kinetic Investigation on Tetrakis(4-Sulfonatophenyl)Porphyrin J-Aggregates Formation Catalyzed by Cationic Metallo-Porphyrins

Under mild acidic conditions, various metal derivatives of tetrakis(4-N-methylpyridinium)porphyrin (gold(III), AuT₄; cobalt(III), CoT₄; manganese(III), MnT₄ and zinc(II), ZnT₄) catalytically promote the supramolecular assembling process of the diacid 5,10,15,20-tetrakis(4-sulfonatophenyl)porphyrin (H₂TPPS₄) into J-aggregates. The aggregation kinetics have been treated according to a well-established model that involves the initial formation of a critical nucleus containing m porphyrin units, followed by autocatalytic growth, in which the rate evolves as a power of time. An analysis of the extinction time traces allows to obtain the rate constants for the auto-catalyzed pathway, k_c, and the number of porphyrins involved in the initial seeding. The aggregation kinetics have been investigated at fixed H₂TPPS₄ concentration as a function of the added metal derivatives MT₄. The derived rate constants, k_c, obey a rate law that is first order in [MT₄] and depend on the specific nature of the catalyst in the order AuT₄ > CoT₄ > MnT₄ > ZnT₄. Both resonance light scattering (RLS) intensity and extinction in the aggregated samples increase on increasing [MT₄]. With the exception of AuT₄, the final aggregated samples obtained at the highest catalyst concentration exhibit a negative Cotton effect in the J-band region, evidencing the occurrence of spontaneous symmetry breaking. The role of the nature of the metal derivative in terms of overall charge and presence of axial groups will be discussed.     

On the functional equation $$f(x+y)=f(x)+f(y)$$f(x+y)=f(x)+f(y)

Monatshefte für Mathematik - An extension of the linearity of the continuous solutions of the functional equation $$f(x+y)=f(x)+f(y)$$ to measurable functions is presented.     

Lipid Nanoparticles Traverse Non-Corneal Path to Reach the Posterior Eye Segment: In Vivo Evidence

Lipid-based nanocarriers (LNs) have made it possible to prolong corneal residence time and improve the ocular bioavailability of ophthalmic drugs. In order to investigate how the LNs interact with the ocular mucosa and reach the posterior eye segment, we have formulated lipid nanocarriers that were designed to bear a traceable fluorescent probe in the present work. The chosen fluorescent probe was obtained by a conjugation reaction between fluoresceinamine and the solid lipid excipient stearic acid, forming a chemically synthesized adduct (ODAF, N-(3′,6′-dihydroxy-3-oxospiro [isobenzofuran-1(3H),9′-[9H] xanthen]-5-yl)-octadecanamide). The novel formulation (LN-ODAF) has been formulated and characterized in terms of its technological parameters (polydispersity index, mean particle size and zeta potential), while an in vivo study was carried out to assess the ability of LN-ODAF to diffuse through different ocular compartments. LN-ODAF were in nanometric range (112.7 nm ± 0.4), showing a good homogeneity and long-term stability. A TEM (transmission electron microscopy) study corroborated these results of characterization. In vivo results pointed out that after ocular instillation, LN ODAF were concentrated in the cornea (two hours), while at a longer time (from the second hour to the eighth hour), the fluorescent signals extended gradually towards the back of the eye. From the results obtained, LN-ODAF demonstrated a potential use of lipid-based nanoparticles as efficient carriers of an active pharmaceutical ingredient (API) involved in the management of retinal diseases.     

Quantum Charges and Spacetime Topology: The Emergence of New Superselection Sectors

A new form of superselection sectors of topological origin is developed. By that it is meant a new investigation that includes several extensions of the traditional framework of Doplicher, Haag and Roberts in local quantum theories. At first we generalize the notion of representations of nets of C*–algebras, then we provide a brand new view on selection criteria by adopting one with a strong topological flavour. We prove that it is coherent with the older point of view, hence a clue to a genuine extension. In this light, we extend Roberts’ cohomological analysis to the case where 1–cocycles bear non-trivial unitary representations of the fundamental group of the spacetime, equivalently of its Cauchy surface in the case of global hyperbolicity. A crucial tool is a notion of group von Neumann algebras generated by the 1–cocycles evaluated on loops over fixed regions. One proves that these group von Neumann algebras are localized at the bounded region where loops start and end and to be factorial of finite type I. All that amounts to a new invariant, in a topological sense, which can be defined as the dimension of the factor. We prove that any 1–cocycle can be factorized into a part that contains only the charge content and another where only the topological information is stored. This second part much resembles what in literature is known as geometric phases. Indeed, by the very geometrical origin of the 1–cocycles that we discuss in the paper, they are essential tools in the theory of net bundles, and the topological part is related to their holonomy content. At the end we prove the existence of net representations. Dedicated to Klaus Fredenhagen on the occasion of his sixtieth birthday     

Catalytic effect of sulfoxides on the fluxional motion of 2,9-dimethyl-1,10-phenanthroline in a platinum(II) complex: evaluation of steric and electronic contributions

Addition of external weak nucleophiles to a chloroform solution of the cationic complex ion [Pt(Me)(dmphen)(PPh(3))](+) (1) accelerates the fluxional motion of the symmetric chelating ligand 2,9-dimethyl-1,10-phenanthroline (dmphen) between nonequivalent exchanging sites. The rates of the dynamic process can be measured by line-shape analysis of the (1)H NMR spectra. Concentration-dependent measurements were carried out with the ligands SOMe(2), SO(CH(2))(4), SO(n-Bu)(2), SO(sec-Bu)(2), SO(i-Pr)(2), SOEt(Ph), SOPr(Ph), SO(Bz)(2), SO(p-MeC(6)H(4))(2), SOPh(2), SO(p-ClC(6)H(4))(2), SOMe(p-MeOC(6)H(4)), SOMe(p-MeC(6)H(4)), SOMe(Ph), SOMe(p-BrC(6)H(4)), and SOMe(p-ClC(6)H(4)). The rate constants k(obsd), when plotted against the concentration of the added ligands SOR(R'), give a family of straight lines with a common intercept, indicating that the two-term rate law k(obsd) = k(1) + k(2)[SOR(R')] is obeyed. The same rate law applies to the displacement of SOMe(2) from [Pt(Me)(phen)(SOMe(2))](+) (2) (phen = 1,10-phenanthroline) by sulfoxides (SOMe(2), SO(i-Pr)(2), SOMe(p-MeOC(6)H(4)), SO(p-MeC(6)H(4))(2), SOPh(2), SO(p-ClC(6)H(4))(2), and SO(sec-Bu)(2)). The fluxional rates in 1 are 6-7 orders of magnitude higher than the substitution rates in 2. The values of the rate constants for the two processes were resolved quantitatively into steric and electronic contributions by use of quantitative analysis of ligand effects (QALE). Inhibitory steric effects are linearly operative for the entire set of ligands, the rates of the reactions are enhanced with increasing electron donor capacity of the sulfoxides, and there is a small but significant E(ar) effect that enhances the reactivity of the aryl sulfoxides. The strict similarity of the patterns of the two processes and of their dependence upon the stereoelectronic properties of the ligands, combined with the intrinsic lability of the platinum-nitrogen bonds, would suggest the operation of stereospecific consecutive ring-opening and ring-closure steps for the fluxional motion of dmphen in 1. However, the available evidence does not allow alternative mechanisms involving intramolecular rearrangements of the five-coordinate intermediate to be ruled out.     

Limits on the simultaneous correlation of gE and hE data by the NRTL,LEMF and Wilson's equations

Three currently popular excess free energy models (Wilson's equation, the NRTL equation, and the LEMF equation) were subjected to a theoretical parametric analysis to determine limits to their ability to correlate experimental g^E and h^E data simultaneously. The LEMF equation was found to be distinctly superior in its ability to predict VLE data from h^E data. Both Wilson's equation and the NRTL equation were shown to break down to ideal solution models in the limit of large intermolecular interactions (|h^E|_max. > 200 cal gmol^?1) whereas the LEMF equation does not. For mixtures whose h^E data exhibit maxima less than 100 cal gmol^?1 and which have positive s^E the LEMF equation coupled with the method of Hanks, Gupta, and Christensen can predict reliable VLE data from h^E data. For |h^E|_max. > 200 cal gmol^?1, the LEMF equation/Hanks—Gupta—Christensen method is accurate to within 10–15% where the other two equations generate errors in excess of 40%.     

Reflection of bounded acoustic beams on a fluid-solid interface

The reflection of an acoustic beam onto a fluid-solid interface is studied under the assumption that the solid medium is viscoelastic. The incident beam is represented as a superposition of plane monochromatic homogeneous waves and its profile is assumed to be Gaussian shaped. The outcoming wave field at the interface, and away from it in the fluid, is numerically calculated for different values of the frequency and of the beam width.

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Sommario Si studia la riflessione di un fascio di onde acustiche sulla superficie di separazione fra un liquido ed un solido nell'ipotesi che il mezzo solido sia viscoelastico. Il fascio incidente ha un profilo gaussiano e viene rappresentato mediante la sovrapposizione di onde piane monocromatiche omogenee. Si calcola numericamente il profilo del fascio riflesso sia sull'interfaccia, sia lontano da essa nel fluido, per diversi valori della frequenza e della larghezza del fascio.

     

Concentration of Bioactive Phenolic Compounds in Olive Mill Wastewater by Direct Contact Membrane Distillation

Olive mill wastewater (OMW), generated as a by-product of olive oil production, is considered one of the most polluting effluents produced by the agro-food industry, due to its high concentration of organic matter and nutrients. However, OMW is rich in several polyphenols, representing compounds with remarkable biological properties. This study aimed to analyze the chemical profile as well as the antioxidant and anti-obesity properties of concentrated fractions obtained from microfiltered OMW treated by direct contact membrane distillation (DCMD). Ultra-high performance liquid chromatography (UHPLC) analyses were applied to quantify some phenols selected as phytochemical markers. Moreover, α-Amylase, α-glucosidase, and lipase inhibitory activity were investigated together with the antioxidant activity by means of assays, namely β-carotene bleaching, 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulphonic) acid (ABTS) diammonium salts, 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay, and Ferric Reducing Activity Power (FRAP) tests. MD retentate—which has content of about five times greater of hydroxytyrosol and verbascoside and about 7 times greater of oleuropein than the feed—was more active as an antioxidant in all applied assays. Of interest is the result obtained in the DPPH test (an inhibitory concentration 50% (IC₅₀) of 9.8 μg/mL in comparison to the feed (IC₅₀ of 97.2 μg/mL)) and in the ABTS assay (an IC₅₀ of 0.4 μg/mL in comparison to the feed (IC₅₀ of 1.2 μg/mL)).