Discovery of Affinity-Based Probes for Btk Occupancy Assays

Bruton's tyrosine kinase (Btk) is an attractive target for the treatment of a wide array of B-cell malignancies and autoimmune diseases. Small-molecule covalent irreversible Btk inhibitors targeting Cys481 have been developed for the treatment of such diseases. In clinical trials, probe molecules are required in occupancy studies to measure the level of engagement of the protein by these covalent irreversible inhibitors. The result of this pharmacodynamic (PD) activity provides guidance for appropriate dosage selection to optimize inhibition of the drug target and correlation of target inhibition with disease treatment efficacy. This information is crucial for successful evaluation of drug candidates in clinical trials. Based on the pyridine carboxamide scaffold of a novel solvent-accessible pocket (SAP) series of covalent irreversible Btk inhibitors, we successfully developed a potent and selective affinity-based biotinylated probe 12 (2-[(4-{4-[5-(1-{5-[(3aS,4S,6aR)-2-oxo-hexahydro-1H-thieno[3,4-d]imidazol-4-yl]pentanamido}-3,6,9,12-tetraoxapentadecan-15-amido)pentanoyl]piperazine-1-carbonyl}phenyl)amino]-6-[1-(prop-2-enoyl)piperidin-4-yl]pyridine-3-carboxamide). Compound 12 has been used in Btk occupancy assays for preclinical studies to determine the therapeutic efficacy of Btk inhibition in two mouse lupus models driven by TLR7 activation and type I interferon.     

Nanoporous artificial proboscis for probing minute amount of liquids

We describe a method of fabrication of nanoporous flexible probes which work as artificial proboscises. The challenge of making probes with fast absorption rates and good retention capacity was addressed theoretically and experimentally. This work shows that the probe should possess two levels of pore hierarchy: nanopores are needed to enhance the capillary action and micrometer pores are required to speed up fluid transport. The model of controlled fluid absorption was verified in experiments. We also demonstrated that the artificial proboscises can be remotely controlled by electric or magnetic fields. Using an artificial proboscis, one can approach a drop of hazardous liquid, absorb it and safely deliver it to an analytical device. With these materials, the paradigm of a stationary microfluidic platform can be shifted to the flexible structures that would allow one to pack multiple microfluidic sensors into a single fiber.     

Enhancement of electrocatalytic properties of carbonized polyaniline nanoparticles upon a hydrothermal treatment in alkaline medium

The electrocatalytic activity of carbonized polyaniline nanostructures (Carb-nanoPANI) towards oxygen reduction reaction (ORR), estimated in 0.1 mol dm⁻³ KOH solution, was significantly improved upon a hydrothermal treatment in 1 mol dm⁻³ KOH solution. Namely, the onset of ORR was shifted by ∼70 mV to more positive potentials, and the number of electrons consumed per O₂ molecule was enhanced in comparison to the original material. The number of electrons involved in ORR depended on loading, and with a loading of 0.5 mg cm⁻², for the potentials lower than −0.5 V vs SCE, the number of electrons approached 4. For this material, high stability of electrochemical behavior and resistance to the poisoning by ethanol was evidenced by potentiodynamic cycling.     

High areal capacitance of FeOOH-carbon nanotube negative electrodes for asymmetric supercapacitors

The relatively low capacitance of negative electrodes, as compared to the capacitance of advanced positive electrodes, poses a serious problem, since this limits the development of asymmetric supercapacitor (SC) devices with a large voltage window and enhanced power-energy characteristics. We fabricate negative SC electrodes with a high capacitance that match the capacitance of advanced positive electrodes at similar active mass loadings, as high as 37?mg?cm^?2. Cyclic voltammetry, impedance spectroscopy, galvanostatic charge-discharge data and the power-energy characteristics of the asymmetric SC device exhibit good electrochemical performance for a voltage window of 1.6?V. Our approach involves the development and application of particle extraction through liquid-liquid interface (PELLI) methods, new extraction mechanisms and efficient extractors to synthesize α-FeOOH and β-FeOOH electrode materials. The use of PELLI allows agglomerate-free processing of powders, which facilitates their efficient mixing with multiwalled carbon nanotubes (MWCNT) and allows improved electrolyte access to the particle surface. Experiments to determine the properties of FeOOH-MWCNT composites provided insight into the influence of the electrode material and the structure of extractor molecules on the composite properties. The highest capacitance of 5.86?F?cm^?2 for negative electrodes and low impedance were achieved using α-FeOOH-MWCNT composites and a 16-phosphonohexadecanoic acid (PHDA) extractor. This extractor allows adsorption on particles, not only at the liquid-liquid interface, but also in the bulk aqueous phase and can potentially be used as a capping agent for particle synthesis and as an extractor in the PELLI method.     

Traveling-wave Phenomena in a Model of Autocrine Signaling Coupled with Dynamics of the MAPK Cascade

Recently we have revealed a minimal reaction subnetwork in the MAPK (mitogen-activated protein kinase) cascade that is responsible for the emergence of bistable and oscillatory behavior. Here we examine a possible mechanism that provides for the propagation of increased MAPK activity in cell populations by interconnecting the intracellular MAPK subnetwork with the ligand-receptor signaling machinery. Such approach allows for significant reduction of the dimensionality of the parameter space on one hand and the conservation of dynamical complexity of the system on the other hand. The coupled model predicts coexistence of one, two or three different stable steady states, or the coexistence of a stable steady state and periodic solution. We found two robust and physiologically relevant characteristics of the proposed model: (i) There is a very large region of coexistence of at least one stable steady state with non-zero MAPK activity and one steady state with zero MAPK activity in the parameter space. (ii) Spontaneous traveling front waves always switching originally inactive cells into ligand releasing cells emerge in adjacent cell populations, e. g. in healthy and injured tissues. Moreover, the formation of composite traveling front waves and spatial oscillatory patterns of MAPK activity are predicted and discussed.     

Nitrogen cycling in tropical grass-legume pastures managed under canopy light interception

Nutrient Cycling in Agroecosystems - In grass-legume pastures, grazing management strategies are an essential factor affecting nitrogen (N) cycling. This study assessed the impact of grazing...     

Modified Low Density Lipoprotein and Lipoprotein-Containing Circulating Immune Complexes as Diagnostic and Prognostic Biomarkers of Atherosclerosis and Type 1 Diabetes Macrovascular Disease

In atherosclerosis; blood low-density lipoproteins (LDL) are subjected to multiple enzymatic and non-enzymatic modifications that increase their atherogenicity and induce immunogenicity. Modified LDL are capable of inducing vascular inflammation through activation of innate immunity; thus, contributing to the progression of atherogenesis. The immunogenicity of modified LDL results in induction of self-antibodies specific to a certain type of modified LDL. The antibodies react with modified LDL forming circulating immune complexes. Circulating immune complexes exhibit prominent immunomodulatory properties that influence atherosclerotic inflammation. Compared to freely circulating modified LDL; modified LDL associated with the immune complexes have a more robust atherogenic and proinflammatory potential. Various lipid components of the immune complexes may serve not only as diagnostic but also as essential predictive markers of cardiovascular events in atherosclerosis. Accumulating evidence indicates that LDL-containing immune complexes can also serve as biomarker for macrovascular disease in type 1 diabetes.     

Possibilities of the Computer-Controlled Detonation Spraying method: A chemistry viewpoint

This article is aimed to discuss the chemical aspects of detonation spraying of powder materials. In this method of coating deposition, ceramic, metallic or composite powders are injected into the barrel of a detonation gun filled with an explosive gaseous mixture. When the latter is ignited, the powders are heated and accelerated toward the substrate. Subjected to high temperatures, the powders are prone to chemical reactions, the reaction products possibly becoming the major phase constituents of the coatings. What types of reactions are possible? Can these reactions be carried out in a controlled manner? We answer these questions considering the interactions of the sprayed powders with the gaseous environment of the barrel as well as those between the phases of a composite feedstock powder. In Computer-Controlled Detonation Spraying (CCDS), the explosive charge and stoichiometry of the fuel-oxygen mixtures are precisely measured and can be flexibly changed. Our studies demonstrate that with the introduction of a highly flexible process of CCDS, detonation spraying has entered a new development stage, at which it can be considered as a powerful method of composition and microstructure tailoring of thermally sprayed coatings. During CCDS of TiO₂-containing powders, chemical reduction of titanium dioxide can be carried out to different levels to form either oxygen-deficient TiO_2−x or Ti₃O₅ suboxide. CCDS of Ti₃Al can produce titanium oxide coatings when oxidation by the detonation products dominates or titanium nitride-titanium aluminide coatings when oxidation is hindered but the interaction of the powders with nitrogen—a carrier gas component—is favored. During detonation spraying of Ti₃SiC₂–Cu composites, the Ti₃SiC₂ phase is preserved only in cold conditions; otherwise, Si de-intercalates from the Ti₃SiC₂ phase and dissolves in Cu resulting in the formation of the TiC_x–Cu(Si) composite coatings.     

The Effect of Titanium, Zirconium and Tin on the Growth of Diatom Synedra Acus and Morphology of Its Silica Valves

The effect of three Group IV metals (titanium, zirconium and tin) on the growth, morphology and chemical composition of the freshwater diatom Synedra acus subsp. radians (Kützing) Skabichevsky was studied and compared with germanium. The elements in their highest oxidation states were introduced into the culture medium in the form of hydroxides. Germanium was found to be toxic at ??5?mol. % of the total Ge-Si content in the culture medium. In the presence of other elements, a slight decrease in the cell division rate was observed independent of the element within 1?C15% content interval. The analysis of the obtained biomass and silica valves revealed the presence of all the added elements within the cells. However, only germanium was incorporated into the valves in considerable amounts. S. acus cultivation with the addition of 5% Group IV elements resulted in cells having the following aberrations in the structure of the silica valves: changes in valve shape, thickening of valves, alterations of the areolae rows, irregularity or absence of the areolae and a decrease in the mechanical strength of valves. Moreover, the effect of Group IV elements on silica formation was simulated in vitro using a synthetic polymer bearing polyamine and phosphate groups found in silaffines (proteins from diatom frustules). The studied elements were observed to provoke the formation of unstable silica particles in solution. We propose that the observed effects of germanium, titanium, zirconium and tin on diatom growth and structure are due to uncontrollable silica condensation.