Spontaneously formed trans-anethol/water/alcohol emulsions: mechanism of formation and stability

We studied the spontaneous emulsification and droplet growth mechanism in trans-anethol/water/ethanol solutions, also known as the beverage ouzo, using dynamic light scattering spectroscopy. This simple ternary mixture is a generic example of a system that forms microemulsions spontaneously when brought into the two-phase region. The volume fraction of the dispersed phase was found to profoundly affect the growth rates of the droplets, which is a new finding that has not been predicted by the Lifshitz-Slyozov-Wagner theory. Time-dependent measurements show that the droplet growth is governed by Ostwald ripening (OR), and no coalescence was observed. Furthermore, the OR rates increase with increasing oil concentration at low alcohol content. We attribute this behavior to enhanced droplet-droplet interactions. At high ethanol concentrations, we found that the measured rates decreased as the oil concentration increased. The OR growth mechanism completely correlates with changes in droplet size. The kinetics of droplet growth shows that the ripening has a saturation limit at a droplet radius of about 1.5 mum. Thus, formed emulsions remain stable for months.     

Reactivity of [Fe4S4(SR)4]2-,3- clusters with sulfonium cations: analogue reaction systems for the initial step in biotin synthase catalysis

The first step in catalysis by a class of iron-sulfur enzymes that includes biotin synthase is the one-electron reductive cleavage of the obligatory cofactor S-adenosylmethionine by an [Fe(4)S(4)](+) cluster to afford methionine and the deoxyadenosyl radical (DOA*). To provide detailed information about the reactions of sulfonium ions with [Fe(4)S(4)](2+,+) clusters, the analogue reaction systems [Fe(4)S(4)(SR')(4)](2)(-)(,3)(-)/[PhMeSCH(2)R](+) (R' = Et (4, 6), Ph (5, 7); R = H (8), COPh (9), p-C(6)H(4)CN (10)) were examined by (1)H NMR spectroscopy. Sulfonium ions 8-10 react completely with oxidized clusters 4 and 5 to afford PhSMe and R'SCH(2)R in equimolar amounts as a result of electrophilic attack by the sulfonium ion on cluster thiolate ligands. Reactions are also complete with reduced clusters 6 and 7 but afford, depending on the substrate, the additional products RCH(3) (R = PhCO, p-C(6)H(4)CN) and the ylid PhMeS=CHR or (p-NCC(6)H(4)CH(2))(2). Redox potentials of 9 and 10 allow electron transfer from 6 or 7. The reaction systems 6/9,10 and 7/9,10 exhibit two reaction pathways, reductive cleavage and electrophilic attack, in an ca. 4:1 ratio inferred from product distribution. Cleavage is a two-electron process and, for example in the system 6/9, is described by the overall reaction 2[Fe(4)S(4)(SR')(4)](3)(-) + 2[PhMeSCH(2)R](+) --> 2[Fe(4)S(4)(SR')(4)](2)(-) + PhSMe + RCH(3) + PhMeS=CHR. This and other reactions may be summarized as [PhMeSCH(2)R](+) + 2e(-) + H(+) --> PhSMe + RCH(3); proposed reaction sequences parallel those for electrochemical reduction of sulfonium ions. This work demonstrates the intrinsic ability of [Fe(4)S(4)](+) clusters with appropriate redox potentials to reductively cleave sulfonium substrates in overall two-electron reactions. The analogue systems differ from the enzymes in that DOA* is generated in a one-electron reduction and is sufficiently stabilized within the protein matrix to abstract a hydrogen atom from substrate or an amino acid residue in a succeeding step. In the present systems, the radical produced in the initial step of the reaction sequence, [Fe(4)S(4)(SR')(4)](3)(-) + [PhMeSCH(2)R](+) --> [Fe(4)S(4)(SR')(4)](2)(-) + PhSMe + RCH(2)*, is not stabilized and is quenched by reduction and protonation.     

Acrolein polymerization: Monodisperse,homo, and hybrido microspheres,synthesis, mechanism,and reactions

Monodisperse polyacrolien (PA) microspheres were obtained by a single step process via two mechanisms: (a) aqueous polymerization of acrolein under alkaline conditions and (b) aqueous radical polymerization of acrolein by irradiation with a cobalt source. The diameter of the former microspheres can be varied from 0.04 up to 8 μm. The monodispersity of the system is also discussed. The diameter of the latter microspheres can be varied up to 0.2 μm. Hybrido PA microspheres were formed by grafting PA microspheres of average diameter of 0.1 μm obtained by irradiation onto the surface of PA microspheres produced by the alkaline mechanism. The aldehyde content of the microspheres prepared by irradiation is much higher than those of the microspheres formed under alkaline conditions. The aldehyde groups were used for the covalent binding of ligands containing primary amino groups, such as proteins and drugs, in a single step under physiological pH.     

Biomineralization of Nickel Struvite Linked to Metal Resistance in Streptomyces mirabilis

Biomineral formation is a common trait and prominent for soil Actinobacteria, including the genus Streptomyces. We investigated the formation of nickel-containing biominerals in the presence of a heavy-metal-resistant Streptomyces mirabilis P16B-1. Biomineralization was found to occur both in solid and liquid media. Minerals were identified with Raman spectroscopy and TEM-EDX to be either Mg-containing struvite produced in media containing no nickel, or Ni-struvite where Ni replaces the Mg when nickel was present in sufficient concentrations in the media. The precipitation of Ni-struvite reduced the concentration of nickel available in the medium. Therefore, Ni-struvite precipitation is an efficient mechanism for tolerance to nickel. We discuss the contribution of a plasmid-encoded nickel efflux transporter in aiding biomineralization. In the elevated local concentrations of Ni surrounding the cells carrying this plasmid, more biominerals occurred supporting this point of view. The biominerals formed have been quantified, showing that the conditions of growth do influence mineralization. This control is also visible in differences observed to biosynthetically synthesized Ni-struvites, including the use of sterile-filtered culture supernatant. The use of the wildtype S. mirabilis P16B-1 and its plasmid-free derivative, as well as a metal-sensitive recipient, S. lividans, and the same transformed with the plasmid, allowed us to access genetic factors involved in this partial control of biomineral formation.     

Synthesis and characterization of an MRI Gd‐based probe designed to target the translocator protein

DPA‐713 is the lead compound of a recently reported pyrazolo[1,5‐a]pyrimidineacetamide series, targeting the translocator protein (TSPO 18 kDa), and as such, this structure, as well as closely related derivatives, have been already successfully used as positron emission tomography radioligands. On the basis of the pharmacological core of this ligands series, a new magnetic resonance imaging probe, coded DPA‐C₆‐(Gd)DOTAMA was designed and successfully synthesized in six steps and 13% overall yield from DPA‐713. The Gd‐DOTA monoamide cage (DOTA = 1,4,7,10‐tetraazacyclododecane‐1,4,7,10‐tetraacetic acid) represents the magnetic resonance imaging reporter, which is spaced from the phenylpyrazolo[1,5‐a]pyrimidineacetamide moiety (DPA‐713 motif) by a six carbon‐atom chain. DPA‐C₆‐(Gd)DOTAMA relaxometric characterization showed the typical behavior of a small‐sized molecule (relaxivity value: 6.02 mM^?1 s^?1 at 20 MHz). The good hydrophilicity of the metal chelate makes DPA‐C₆‐(Gd)DOTAMA soluble in water, affecting thus its biodistribution with respect to the parent lipophilic DPA‐713 molecule. For this reason, it was deemed of interest to load the probe to a large carrier in order to increase its residence lifetime in blood. Whereas DPA‐C₆‐(Gd)DOTAMA binds to serum albumin with a low affinity constant, it can be entrapped into liposomes (both in the membrane and in the inner aqueous cavity). The stability of the supramolecular adduct formed by the Gd‐complex and liposomes was assessed by a competition test with albumin. Copyright © 2013 John Wiley & Sons, Ltd.     

Rutin and Quercetin Decrease Cholesterol in HepG2 Cells but Not Plasma Cholesterol in Hamsters by Oral Administration

Rutin (R) and quercetin (Q) are two widespread dietary flavonoids. Previous studies regarding the plasma cholesterol-lowering activity of R and Q generated inconsistent results. The present study was therefore carried out to investigate the effects of R and Q on cholesterol metabolism in both HepG2 cells and hypercholesterolemia hamsters. Results from HepG2 cell experiments demonstrate that both R and Q decreased cholesterol at doses of 5 and 10 µM. R and Q up-regulated both the mRNA and protein expression of sterol regulatory element binding protein 2 (SREBP2), low-density lipoprotein receptor (LDLR), and liver X receptor alpha (LXRα). The immunofluorescence study revealed that R and Q increased the LDLR expression, while only Q improved LDL-C uptake in HepG2 cells. Results from hypercholesterolemia hamsters fed diets containing R (5.5 g/kg diet) and Q (2.5 g/kg diet) for 8 weeks demonstrate that both R and Q had no effect on plasma total cholesterol. In the liver, only Q reduced cholesterol significantly. The discrepancy between the in vitro and in vivo studies was probably due to a poor bioavailability of flavonoids in the intestine. It was therefore concluded that R and Q were effective in reducing cholesterol in HepG2 cells in vitro, whereas in vivo, the oral administration of the two flavonoids had little effect on plasma cholesterol in hamsters.     

Design,Synthesis and Preclinical Assessment of 99mTc-iFAP for In Vivo Fibroblast Activation Protein (FAP) Imaging

Fibroblast activation protein (FAP) is expressed in the microenvironment of most human epithelial tumors. ⁶⁸Ga-labeled FAP inhibitors based on the cyanopyrrolidine structure (FAPI) are currently used for the detection of the tumor microenvironment by PET imaging. This research aimed to design, synthesize and preclinically evaluate a new FAP inhibitor radiopharmaceutical based on the ^99mTc-((R)-1-((6-hydrazinylnicotinoyl)-D-alanyl) pyrrolidin-2-yl) boronic acid (^99mTc-iFAP) structure for SPECT imaging. Molecular docking for affinity calculations was performed using the AutoDock software. The chemical synthesis was based on a series of coupling reactions of 6-hidrazinylnicotinic acid (HYNIC) and D-alanine to a boronic acid derivative. The iFAP was prepared as a lyophilized formulation based on EDDA/SnCl₂ for labeling with ^99mTc. The radiochemical purity (R.P.) was verified via ITLC-SG and reversed-phase radio-HPLC. The stability in human serum was evaluated by size-exclusion HPLC. In vitro cell uptake was assessed using N30 stromal endometrial cells (FAP positive) and human fibroblasts (FAP negative). Biodistribution and tumor uptake were determined in Hep-G2 tumor-bearing nude mice, from which images were acquired using a micro-SPECT/CT. The iFAP ligand (Ki = 0.536 nm, AutoDock affinity), characterized by UV-Vis, FT-IR, ¹H–NMR and UPLC-mass spectroscopies, was synthesized with a chemical purity of 92%. The ^99mTc-iFAP was obtained with a R.P. >98%. In vitro and in vivo studies indicated high radiotracer stability in human serum (>95% at 24 h), specific recognition for FAP, high tumor uptake (7.05 ± 1.13% ID/g at 30 min) and fast kidney elimination. The results found in this research justify additional dosimetric and clinical studies to establish the sensitivity and specificity of the ^99mTc-iFAP.     

Maximum confidence quantum measurements

We consider the problem of discriminating between states of a specified set with maximum confidence. For a set of linearly independent states unambiguous discrimination is possible if we allow for the possibility of an inconclusive result. For linearly dependent sets an analogous measurement is one which allows us to be as confident as possible that when a given state is identified on the basis of the measurement result, it is indeed the correct state.     

Effect of temperature on the synthesis of silver nanoparticles with polyethylene glycol: new insights into the reduction mechanism

Polyethylene glycol (PEG) molecules act as a reducing and stabilizing agent in the formation of silver nanoparticles. PEG undergoes thermal oxidative degradation at temperatures over 70 °C in the presence of oxygen. Here, we studied how the temperature and an oxidizing atmosphere could affect the synthesis of silver nanoparticles with PEG. We tested different AgNO₃ concentrations for nanoparticles syntheses using PEG of low molecular weight, at 60 and 100 °C. At the higher temperature, the reducing action of PEG increased and the effect of PEG/Ag⁺ ratio on nanoparticles aggregation changed. These results suggest that different synthesis mechanisms operate at 60 and 100 °C. Thus, at 60 °C the reduction of silver ions can occur through the oxidation of the hydroxyl groups of PEG, as has been previously reported. We propose that the thermal oxidative degradation of PEG at 100 °C increases the number of both, functional groups and molecules that can reduce silver ions and stabilize silver nanoparticles. This degradation process could explain the enhancement of PEG reducing action observed by other authors when they increase the reaction temperature or use a PEG of higher molecular weight