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.     

Liquid phase sintering and characterization of SiC ceramics

The present study focuses on the sintering of silicon carbide-based ceramics (SiC) by liquid phase sintering (LPS) followed by characterization of the produced ceramics. AlN/Re₂O₃ mixtures were used as additives in the LPS process. In the first step, the LPS-SiC materials were produced in a graphite resistance furnace in the form of discs at different temperatures. The conditions with the best results regarding real density and relative density were taken as reference for sintering in the form of prismatic bars. In the second step, these samples were evaluated regarding fracture toughness (K_IC), by the Single Edge V Notch Beam – SEVNB – method, and flexural strength. K_IC behavior was evaluated according to the depth and curvature radius of the notches. Reliable K_IC values were presented when the ceramic displayed a small curvature radius at the notch tip. When the radius was large, it did not maintain the square root singularity of the notch tip. Tests were carried out to determine K_IC values in atmospheric air and water. K_IC results were lower in water than air, with a decrease ranging between 2.56% and 11.26%. The observations indicated a direct grain size correlation between K_IC values and fracture strength of the SiC ceramics.     

A Constitutive Equation of Thermoelasticity for Solid Materials

A new constitutive equation of thermoelasticity for crystals is presented based on the interatomic potential and solid mechanics at finite temperature. Using the new constitutive equation, the calculations for crystal copper and graphene are carried out under different loading paths at different temperatures. The calculated results are in good agreement with those of the previous thermoelasticity constitutive equation based on quantum mechanics, which clearly indicates that our new constitutive equation of thermoelasticity is correct. A lot of comparisons also show that the present theory is more concise and efficient than the previous thermal stress theory in the practical application.     

Nutrient digestibility and endogenous protein losses in the foregut and small intestine of weaned dairy calves fed calf starters with conventional or enzyme-treated soybean meal

The aims of this experiment were (1) to compare the effects of a soybean meal with an enzymatic treatment (ESBM) to reduce the concentration of antinutritional factors versus a standard soybean meal (SBM) on foregut and small intestine digestion in weaned dairy calves and (2) to estimate the endogenous losses of crude protein (CP) in the small intestine. Our hypothesis was that a diet containing ESBM instead of SBM would improve ruminal and small intestine digestion and absorption of nutrients. A T-cannula was placed in the duodenum, and a second T-cannula was installed in the distal ileum of 12 Holstein calves at approximately 3 wk of age. Calves were weaned on d 42, and on d 50 they were assigned randomly to a quadruplicated 3 × 3 Latin square with 10-d periods. Digesta samples were collected on d 7 and 8 from the ileum and d 9 and 10 from the duodenum. The diets were fed for ad libitum intake and consisted of a calf starter (CS) of 20% CP with SBM as the main source of protein (CTRL), and an isonitrogenous CS with an ESBM instead of SBM (ENZT). A third diet with a low content of CP (10%) and no soy protein was fed to estimate endogenous N losses and digestibilities of test ingredients. Flows and digestibilities of nutrients were compared between CTRL and ENZT and their test ingredients (SBM vs. ESBM, respectively). Duodenal net flows of CP and total AA as well as ruminal microbial protein synthesis per kilogram of digested CP were greater, and flow of nonprotein N and CP true (corrected by endogenous and microbial flows) foregut digestibility were lower with ENZT than CTRL. The apparent small intestine digestibilities of CP and total AA were greater for ESBM than SBM, but there were no differences between the CTRL and ENZT diets. We observed no differences in digestibilities at the duodenum or ileum of starch or NDF, but true small intestine digestibilities of CP and all AA were greater with ENZT than CTRL. Total endogenous protein losses in the small intestine estimated from calves fed the low-CP with no soy protein diet were 37 ± 1.5 g of CP and 29 ± 1.4 g of AA/kg of DMI. These values may be considered the basal endogenous losses as they are similar to values obtained with the regression method, which estimates N losses when dietary N is null. Our results indicated that the inclusion of an ESBM improved the efficiency of ruminal microbial protein synthesis per digested kilogram of organic matter and CP, and increased CP and AA absorption in the small intestine despite a greater proportion of undigested dietary protein entering the duodenum.     

Thermal Stability of Polymer Additives: Comparison of Decomposition Models Including Oxidative Pyrolysis

The thermo‐oxidative stability of widely used polymer additives has been investigated. A comparative analytical approach with classic and innovative decomposition models for polymer additives was conducted and the results supported using quantum‐chemical calculations. Unique pyrolysis products of the analytes were compiled utilizing pyrolysis online coupled to gas chromatography followed by mass spectrometric detection (Pyr‐GC–MS). The pyrolysis was either performed under inert conditions or in an oxygen‐containing atmosphere. Squalane was applied as polymer‐mimicking liquid next to low density polyethylene (LDPE) and polyamide 6 (PA 6) as matrices for 10 selected additives. The additives included in this study range from antioxidants and plasticizers to processing aids. These were selected to address a range of application in consumer products and to cover different chemical classes. The toxicological relevance of additives and potential breakdown products was considered. Consequently, degradation of sterically hindered antioxidants, diarylamines, and a trimellitic acid derivative was investigated. The findings were used to predict the behavior of consumer products made of polymeric materials entailing additives. The level of Antioxidant 2246 [2‐tert‐butyl‐6‐[(3‐tert‐butyl‐2‐hydroxy‐5‐methylphenyl)methyl]‐4‐methylphenol] and one of its predicted decomposition products was determined in baby bottle nipples made of natural rubber [2‐tert‐butyl‐4‐methylphenol] utilizing the complementary technique of gas chromatography coupled to tandem mass spectrometry (GC–MS/MS). This study provides a comprehensive characterization of important polymer additives and enables the prioritization of degradation products for further risk assessment. J. VINYL ADDIT. TECHNOL., 25:E12–E27, 2019. © 2018 The Authors. Journal of Vinyl and Additive Technology published by Wiley Periodicals, Inc. on behalf of Society of Plastics Engineers     

Calcium phosphates grown on bacterial cellulose template

Bacterial cellulose membranes were employed as templates for calcium phosphates deposition by successive immersion in solutions of Ca(NO₃)₂·4H₂O and (NH₄)₂HPO₄, under ultrasonication. During the wet chemical reaction, mineral phases were loaded on bacterial cellulose fibrils, leading to precursor hybrid composites. These were subjected to a lyophilisation procedure in order to preserve the 3D porous aspect and afterwards to a thermal treatment with the aim of removing the polymeric phase and generating well crystallized structures. Different types of morphologies were achieved by varying the heating rate, as well as the calcination temperature and period. The as-prepared samples and the final ones were investigated from compositional and structural point of view through X-ray diffraction and Fourier-transform infrared spectroscopy and morphologically concerning by scanning electron microscopy. The magnetic properties were also evaluated in order to demonstrate the suitability of the obtained materials for the development of magnetic scaffolds dedicated to hard tissue applications.