Novel Redox-Dependent Esterase Activity (EC 3.1.1.2) for DJ-1: Implications for Parkinson’s Disease

Mutations the in human DJ-1 (hDJ-1) gene are associated with early-onset autosomal recessive forms of Parkinson’s disease (PD). hDJ-1/parkinsonism associated deglycase (PARK7) is a cytoprotective multi-functional protein that contains a conserved cysteine-protease domain. Given that cysteine-proteases can act on both amide and ester substrates, we surmised that hDJ-1 possessed cysteine-mediated esterase activity. To test this hypothesis, hDJ-1 was overexpressed, purified and tested for activity towards 4-nitrophenyl acetate (pNPA) as µmol of pNPA hydrolyzed/min/mg·protein (U/mg protein). hDJ-1 showed maximum reaction velocity esterase activity (V_max = 235.10 ± 12.00 U/mg protein), with a sigmoidal fit (S_0.5 = 0.55 ± 0.040 mM) and apparent positive cooperativity (Hill coefficient of 2.05 ± 0.28). A PD-associated mutant of DJ-1 (M26I) lacked activity. Unlike its protease activity which is inactivated by reactive oxygen species (ROS), esterase activity of hDJ-1 is enhanced upon exposure to low concentrations of hydrogen peroxide (<10 µM) and plateaus at elevated concentrations (>100 µM) suggesting that its activity is resistant to oxidative stress. Esterase activity of DJ-1 requires oxidation of catalytic cysteines, as chemically protecting cysteines blocked its activity whereas an oxido-mimetic mutant of DJ-1 (C106D) exhibited robust esterase activity. Molecular docking studies suggest that C106 and L126 within its catalytic site interact with esterase substrates. Overall, our data show that hDJ-1 contains intrinsic redox-sensitive esterase activity that is abolished in a PD-associated mutant form of the hDJ-1 protein.     

Pentadecane functionalized graphite oxide sheets as a tool for the preparation of electrical conductive polyethylene/graphite oxide composites

Covalent functionalization of pentadecane-decorated thermally reduced graphite oxide (GO) sheets has been studied as a tool for the preparation of polyethylene/GO composites exhibiting rheological and electrical percolation thresholds. It was accomplished through pentadecane based radical addition onto unsaturated bonds located on the GO sheets' surface using dicumyl peroxide as hydrogen abstractor. This chemical functionalization influences the affinity of the formed pentadecane grafted GO sheets for various solvents. Then, the compounding of the composites pentadecane grafted GO/PE was performed at a processing temperature of 140 °C with 25, 20, 15, 10, 8 and 5 wt% loadings. Rheological and electrical percolation thresholds were found between 10 and 15 wt% for polyethylene/pentadecane functionalized graphene oxide composites while the composite graphite/PE at the same loading percentage did not reach any percolation threshold.     

Defect structure evolution and electrical properties of BaTiO3-based ferroelectric ceramics

Relaxor perovskite ferroelectric 0.1Bi(Zn_1/2Zr_1/2)O₃-0.9BaTiO₃(0.1BZZ-0.9BT) ceramics were successfully prepared, whose powders synthesized by the sol-gel process, with average grain size about 1.29 μm. 1.75 J/cm³ discharge energy density and good dielectric stability were obtained over a wide temperature range from 25°C to 140°C. The pulse discharge capability of 0.1BZZ-0.9BT ceramics was tested under different electric fields. The discharge time was 2.13 μs, which proved its ability to charge and discharge quickly. Complex impedance analysis and thermally stimulated depolarization current tests were applied to investigate the defect types and activation of 0.1BZZ-0.9BT ceramics. The evolution process of composite defects and oxygen vacancies profoundly affects the dielectric temperature stability of 0.1BZZ-0.9BT ceramics’ energy storage property.     

Targeting Casein Kinase 1 (CK1) in Hematological Cancers

The casein kinase 1 enzymes (CK1) form a family of serine/threonine kinases with seven CK1 isoforms identified in humans. The most important substrates of CK1 kinases are proteins that act in the regulatory nodes essential for tumorigenesis of hematological malignancies. Among those, the most important are the functions of CK1s in the regulation of Wnt pathways, cell proliferation, apoptosis and autophagy. In this review we summarize the recent developments in the understanding of biology and therapeutic potential of the inhibition of CK1 isoforms in the pathogenesis of chronic lymphocytic leukemia (CLL), other non-Hodgkin lymphomas (NHL), myelodysplastic syndrome (MDS), acute myeloid leukemia (AML) and multiple myeloma (MM). CK1δ/ε inhibitors block CLL development in preclinical models via inhibition of WNT-5A/ROR1-driven non-canonical Wnt pathway. While no selective CK1 inhibitors have reached clinical stage to date, one dual PI3Kδ and CK1ε inhibitor, umbralisib, is currently in clinical trials for CLL and NHL patients. In MDS, AML and MM, inhibition of CK1α, acting via activation of p53 pathway, showed promising preclinical activities and the first CK1α inhibitor has now entered the clinical trials.     

Polysilazane derived micro/nano Si3N4/SiC composites

The pyrolised polysilazanes poly(hydridomethyl)silazane NCP 200 and poly(urea)silazane CERASET derived Si–C–N amorphous powders were used for preparation of micro/nano Si₃N₄/SiC composites by hot pressing. Y₂O₃–Al₂O₃ and Y₂O₃–Yb₂O₃ were used, as sintering aids. The resulting ceramic composites of all compositions were dense and polycrystalline with fine microstructure of average grain size <1 μm of both Si₃N₄ and SiC phases. The fine SiC nano-inclusions were identified within the Si₃N₄ micrograins. Phase composition of both composites consist of , β modifications of Si₃N₄ and SiC. High weight loss was observed during the hot pressing cycle, 12 and 19 wt.% for NCP 200 and CERASET precursors, respectively. The fracture toughness of both nanocomposites (NCP 2000 and CERASET derived) was not different. Indentation method measured values are from 5 to 6 MPa m^1/2, with respect to the sintering additive system. Fracture toughness is slightly sensitive to the SiC content of the nanocomposite. Hardness increases with the content of SiC in the nanocomposite. The highest hardness was achieved for pyrolysed CERASET precursor with 2 wt.% Y₂O₃ and 6 wt.% Yb₂O₃, HV 23 GPa. This is a consequence of the highest SiC content as well as the chemical composition of additives.     

Generation of spatio-temporal concentration profiles for cell culture systems: A case study in ammonia

Cellular behavior is influenced by micro-concentration gradients existing in their immediate spatio-environment. A practical real-time measurement system, composed of miniaturized long-lasting sensors placed within a two-dimensional bioreactor and a 32-channel data acquisition system for ammonia, is presented in this paper aiming to generate the spatio-temporal ammonia concentration profiles for the whole culture process. Our results demonstrate the feasibility of the system and its applicability in elucidating the cell growth micro-environment within a bioreactor, which can potentially facilitate the intelligent control of ammonia levels in cell culture systems. The presented system is flexible enough to interface other sensors for generating their spatio-temporal profiles of the sensed cell culture parameters.     

Development and Characterization of Continuous SiC Fiber‐Reinforced HfB2‐Based UHTC Matrix Composites Using Polymer Impregnation and Slurry Infiltration Techniques

This paper discusses the development of continuous SiC fiber‐reinforced HfB₂‐SiC composite laminates. A range of techniques, based on resin‐based precursors and slurries, for infiltrating porous SiC preforms with HfB₂ powder were developed. While resin‐based precursors proved to be ineffective due to low HfB₂ yield and poor adhesion, the slurry infiltration techniques were effective to varying degrees. The greatest pore filling and composite densities were achieved using pressure and vibration‐assisted pressure infiltration techniques. SiC_f/HfB₂‐SiC laminates were subsequently developed via lamination, cure and pyrolysis of fabrics using a HfB₂‐loaded polymeric SiC precursor, followed by HfB₂ slurry infiltration and preceramic polymer infiltration and pyrolysis (PIP). Repeated PIP processing, for 6–10 cycles, resulted in density increases, from the 3.03–3.22 g/cm³ range after HfB₂ slurry infiltration, to 3.97–4.03 g/cm³ after PIP processing. Correspondingly, there was a decrease in open porosity from approximately 52% to less than 11%. The matrix consisted of discreet, lightly sintered HfB₂ particles dispersed in SiC. The PIP SiC matrix was primarily nanocrystalline after 1300°C pyrolysis, but experienced grain growth with further heat treatment at 1600°C.     

Extraction of oleic acid from soybean oil and jojoba oil-phase diagrams

Formamide,N-methylformamide,N-dimethylformamide, dimethylsulfoxide, 1,2-butanediol, and 2-butane-1,4-diol were considered as potential extractants of fatty acids from soybean and jojoba oils. Ternary liquid-liquid phase diagrams at 298.15 K, distribution, and selectivity coefficients of oleic acid are reported. Of the investigated solvents, onlyN-methylformamide and 1,2-butanediol have desirable extraction characteristics.     

An end‐to‐end alternative to TCP PEPs: Initial Spreading,a TCP fast start‐up mechanism

While Transmission Control Protocol (TCP) Performance Enhancing Proxy (PEP) solutions have long been undisputed to solve the inherent satellite problems, the improvement of the regular end‐to‐end TCP congestion avoidance algorithms and the recent emphasis on the PEPs drawbacks have opened the question of the PEPs sustainability. Nevertheless, with a vast majority of Internet connections shorter than ten segments, TCP PEPs continue to be required to counter the poor efficiency of the end‐to‐end TCP start‐up mechanisms. To reduce the PEPs dependency, designing a new fast start‐up TCP mechanism is therefore a major concern. But, while enlarging the Initial Window (IW) up to ten segments is, without any doubt, the fastest solution to deal with a short‐lived connection in an uncongested network, numerous researchers are concerned about the impact of the large initial burst on an already congested network. Based on traffic observations and real experiments, Initial Spreading has been designed to remove those concerns whatever the load and type of networks. It offers performance similar to a large IW in uncongested network and outperforms existing end‐to‐end solutions in congested networks. In this paper, we show that Initial Spreading, taking care of the satellite specificities, is an efficient end‐to‐end alternative to the TCP PEPs. Copyright © 2015 John Wiley & Sons, Ltd.