Janus‐Like Squaramide‐Based Hosts: Dual Mode of Binding and Conformational Transitions Driven by Ion‐Pair Recognition

New tripodal squaramide‐based hosts have been synthesised and structurally characterised by spectroscopic methods. In 2.5 % (v/v) [D₆]DMSO in CDCl₃, compound 4 formed dimeric assemblies [log K_dim=3.68(8)] as demonstrated by ¹H NMR spectroscopy and UV dilution experiments. AFM and SEM analyses revealed the formation of a network of bundled fibres, which indicates a preferential mechanism for aggregation. These C₃‐symmetric tripodal hosts exhibited two different and mutually exclusive modes of binding, each one easily accessible by simultaneous reorientation of the squaramide groups. In the first, a convergent disposition of the NH squaramide protons allowed the formation of an array of N? H???X^? hydrogen bonds with anions. In the second mode, reorientation of carbonyl squaramide groups allowed multiple C?O???H interactions with ammonium cations. The titration of 4 with different tetraalkylammonium iodides persistently showed the formation of 1:1 complexes, as well as 1:2 and 1:3 complexes. The corresponding stoichiometries and binding affinities of the complexes were evaluated by multi‐regression analysis. The formation of high‐order complexes, supported by ROESY, NOESY and mass spectrometry experiments, has been attributed to the insertion of NR₄I ion pairs between the carbonyl and NH protons of the squaramide groups located in adjacent arms of 4 . The observed effects reflect the induction of significant conformational changes in the hosts, mainly in relation to the relative orientation of the squaramide groups adapting their geometries to incoming ion‐pair complementary substrates. The results presented herein identify and fully describe two different modes of ion‐pair recognition aimed at directing conformational transitions in the host, therefore establishing a base for controlling more elaborate movements of molecular devices through ion‐pair recognition.     

Substrate Range of the Titanium TADDOLate Catalyzed Asymmetric Fluorination of Activated Carbonyl Compounds

The substrate range of the [TiCl₂(TADDOLate)] (TADDOL=α,α,α′,α′‐tetraaryl‐1,3‐dioxolane‐4,5‐dimethanol)‐catalyzed asymmetric α‐fluorination of activated β‐carbonyl compounds has been investigated. Optimal conditions for catalysis are characterized by using 5 mol‐% of TiCl₂(naphthalen‐1‐yl)‐TADDOLate) as catalyst in a saturated (0.14 mol/l) MeCN solution of F‐TEDA (1‐(chloromethyl)‐4‐fluoro‐1,4‐diazoniabicyclo[2.2.2]octane bis‐[tetrafluoroborate]) at room temperature. A series of α‐methylated β‐keto esters (3‐oxobutanoates, 3‐oxopentanoates) with bulky benzyl ester groups (60–90% ee) or phenyl ester (67–88% ee) have been fluorinated readily, whereas α‐acyl lactones were also readily fluorinated, but gave lower inductions (13–46% ee). Double stereochemical differentiation in β‐keto esters with chiral ester groups raised the stereoselectivity to a diastereomeric ratio (dr) of up to 96.5 : 3.5. For the first time, β‐keto S‐thioesters were asymmetrically fluorinated (62–91.5% ee) and chlorinated (83% ee). Lower inductions were observed in fluorinations of 1,3‐diketones (up to 40% ee) and β‐keto amides (up to 59% ee). General strategies for preparing activated β‐carbonyl compounds as important model substrates for asymmetric catalytic α‐functionalizations are presented (>60 examples).     

Anti-inflammatory activity of Pistacia lentiscus essential oil: involvement of IL-6 and TNF-alpha

The topical anti-inflammatory activity of essential oil of Pistacia lentiscus L. was studied using carrageenan induced rat paw edema and cotton pellet induced granuloma. The effect on serum tumor necrosis factor-alpha (TNF-alpha) and interleukin-6 (IL-6) in rats inserted with cotton pellet was also investigated. On topical application, the oil exhibited a significant decrease in paw edema. The oil also inhibited cotton pellet-induced granuloma, and reduced serum TNF-alpha and IL-6. It can be concluded that the essential oil of Pistacia lentiscus reduces leukocyte migration to the damaged tissue and exhibits anti-inflammatory activity.     

Detection of DNA hybridisation in a diluted serum matrix by surface plasmon resonance and film bulk acoustic resonators

Nanomolar quantities of single-stranded DNA products ~100 nucleotides long can be detected in diluted 1% serum by surface plasmon resonance (SPR) and film bulk acoustic resonators (FBARs). We have used a novel FBAR sensor in parallel with SPR and obtained promising results with both the acoustic and the optical device. Oligonucleotides and a repellent lipoamide, Lipa-DEA, were allowed to assemble on the sensor chip surfaces for only 15 min by dispensing. Lipa-DEA surrounds the analyte-binding probes on the surface and effectively reduces the non-specific binding of bovine serum albumin and non-complementary strands. In a highly diluted serum matrix, the non-specific binding is, however, a hindrance, and the background response must be reduced. Nanomolar concentrations of short complementary oligos could be detected in buffer, whereas the response was too low to be measured in serum. DNA strands that are approximately 100 base pairs long at concentrations as low as 1-nM could be detected both in buffer and in 1% serum by both SPR and the FBAR resonator.     

Determination of homocysteine thiolactone, reduced homocysteine, homocystine, homocysteine-cysteine mixed disulfide, cysteine and cystine in a reaction mixture by overimposed pressure/voltage capillary electrophoresis

An elevated level of thiol amino acid homocysteine is associated with several complex disorders. Homocysteine ability to bind proteins, thereby modulating their structure and function, is proposed to be one of the mechanisms of homocysteine induced pathogenecity. Homocysteine and homocysteine thiolactone bind to protein cysteine and lysine residues respectively. A major hurdle in studying protein homocysteinylation is the lack of suitable analytical techniques to determine simultaneously the concentrations of reduced and oxidized forms of homocysteine and cysteine (especially homocysteine-cysteine mixed disulfide) together with thiolactone formed during the reaction of homocysteine or thiolactone with proteins. Herein we report a capillary electrophoresis method to determine simultaneously the levels of these intermediates. For this 40 mmol/L Tris phosphate buffer at (pH 1.60) was used as running electrolyte, and the separation was performed by the simultaneous application of a CE voltage of 15 kV and an overimposed pressure of 0.1 psi. A rapid separation of these intermediates in less than 6 min with a good reproducibility of both peak areas (CV < 2%) and migration time (CV < 0.2%) was obtained. The applicability of our method was validated by incubating reduced homocysteine and albumin and measuring the reaction intermediates in the solution mixture.     

Inhibition of histamine mediated responses by Mirabilis jalapa: confirming traditional claims made about antiallergic and antiasthmatic activity

The roots of Mirabilis jalapa are used traditionally in allergic skin disorders and asthma. The effect of an ethanol:acetone (1:1) extract of the roots of M. jalapa was studied for antihistaminic activity using a guinea pig tracheal chain preparation and clonidine-induced mast cell granulation in mice. Its antiallergic activity was evaluated using milk-induced eosinophilia and albumin-induced paw oedema in mice. The extract (0.5 mL of 100 mg mL(-1)) inhibited histamine-induced guinea pig tracheal chain contractions non-competitively. The extract (100 or 200 mg kg(-1) i.p.) inhibited milk-induced eosinophilia, albumin-induced paw oedema and protected mast cells against clonidine-induced granulation. The study justified the folkloric use of M. jalapa in the treatment of allergic diseases and asthma.     

Quantification of histidine, 1-methylhistidine and 3-methylhistidine in plasma and urine by capillary electrophoresis UV-detection

We describe a new CE method with UV-detection for the quantification of histidine (His) and its methylated forms 1-methylhistidine and 3-methylhistidine, both in plasma and urine. Analytes were basically resolved using a 60?mmol/L Tris-phosphate run buffer pH 2.2 in less than 12?min. The use of a mixture of ACN/ammonia (80:20) for protein precipitation allows the quantitative recovery of all His from plasma. The optimization of the sample volume injection permits to reach an LOD of 20?nmol/L, thus improving the sensitivity of about hundred times in comparison to the previous described assays. Moreover, the opportunity to also measure creatinine in the same run makes it possible to evaluate the renal function contemporarily, thus avoiding further dosages with significant time saving. The application method has been proved by measuring His, 1-methylhistidine and 3-methylhistidine in 44 healthy subjects. In conclusion, our new method seems to be an inexpensive, fast and specific tool to assess large numbers of patients for routine analysis both in clinical and research laboratories.     

VO2+ complexation by bioligands showing keto-enol tautomerism: a potentiometric, spectroscopic, and computational study

The interaction of VO(2+) ion with ligands of biological interest that are present in important metabolic pathways--2-oxopropanoic acid (pyruvic acid, pyrH), 3-hydroxy-2-oxopropanoic acid (3-hydroxypyruvic acid, hydpyrH), oxobutanedioic acid (oxalacetic acid, oxalH(2)), (S)-hydroxybutanedioic acid (l-malic acid, malH(2)), and 2,3-dihydroxy-(E)-butanedioic acid (dihydroxyfumaric acid, dhfH(2))--was described. Their complexing capability was compared with that of similar ligands: 3-hydroxy-2-butanone (hydbut) and 3,4-dihydroxy-3-cyclobutene-1,2-dione (squaric acid, squarH(2)). All of these ligands (except l-malic acid) exhibit keto-enol tautomerism, and the presence of a metal ion can influence such an equilibrium. The different systems were studied with electron paramagnetic resonance (EPR) and UV-vis spectroscopies and with pH potentiometry. Density functional theory (DFT) methods provide valuable information on the relative energy of the enol and keto forms of the ligands both in the gas phase and in aqueous solution, on the geometry of the complexes, and on EPR and electronic absorption parameters. The results show that most of the ligands behave like α-hydroxycarboxylates, forming mono- and bis-chelated species with (COO(-), O(-)) coordination, demonstrating that the metal ion is able to stabilize the enolate form of some ligands. With dihydroxyfumaric acid, the formation of a non-oxidovanadium(IV) complex, because of rearrangement of dihydroxyfumaric to dihydroxymaleic acid (dhmH(2)), can be observed. With 3-hydroxy-2-butanone and 3,4-dihydroxy-3-cyclobutene-1,2-dione, complexation of VO(2+) does not take place and the reason for this behavior is explained by chemical considerations and computational calculations.     

The effect of wall depletion on the rheology of microfibrillated cellulose water suspensions by optical coherence tomography

Rheology of microfibrillated cellulose (MFC) water suspensions was characterized with a rotational rheometer, augmented with optical coherence tomography (OCT). To the best of the authors’ knowledge, this is the first time the behavior of MFC in the rheometer gap was characterized by this real-time imaging method. Two concentrations, 0.5 and 1 wt% were used, the latter also with 10^?3 and 10^?2 M NaCl. The aim was to follow the structure of the suspensions in a rotational rheometer during the measurements and observe wall depletion and other factors that can interfere with the rheological results. The stepped flow measurements were performed using a transparent cylindrical measuring system and combining the optical information to rheological parameters. OCT allows imaging in radial direction from the outer geometry boundary to the inner geometry boundary making both the shear rate profile and the structure of the suspension visible through the rheometer gap. Yield stress and maximum wall stress were determined by start-up of steady shear and logarithmic stress ramp methods and they both reflected in the stepped flow measurements. Above yield stress, floc size was inversely proportional to shear rate. Below the yield stress, flocs adhered to each other and the observed apparent constant shear stress was controlled by flow in the depleted boundary layer. With higher ionic strength (10^?2 M NaCl), the combination of yield stress and wall depletion favored the formation of vertical, cylindrical, rotating floc structures (rollers) coupled with a thicker water layer originating at the suspension—inner cylinder boundary at low shear rates.