Discovery of Five New Ethylene-Forming Enzymes for Clean Production of Ethylene in E. coli

Ethylene is an essential platform chemical with a conjugated double bond, which can produce many secondary chemical products through copolymerisation. At present, ethylene production is mainly from petroleum fractionation and cracking, which are unsustainable in the long term, and harmful to our environment. Therefore, a hot research field is seeking a cleaner method for ethylene production. Based on the model ethylene-forming enzyme (Efe) {"type":"entrez-protein","attrs":{"text":"AAD16440.1","term_id":"4323597","term_text":"AAD16440.1"}}AAD16440.1 (6vp4.1.A) from Pseudomonas syringae pv. phaseolicol, we evaluated five putative Efe protein sequences using the data derived from phylogenetic analyses and the conservation of their catalytic structures. Then, pBAD expression frameworks were constructed, and relevant enzymes were expressed in E. coli BL21. Finally, enzymatic activity in vitro and in vivo was detected to demonstrate their catalytic activity. Our results show that the activity in vitro measured by the conversion of α-ketoglutarate was from 0.21–0.72 μmol ethylene/mg/min, which varied across the temperatures. In cells, the activity of the new Efes was 12.28–147.43 μmol/gDCW/h (DCW, dry cellular weight). Both results prove that all the five putative Efes could produce ethylene.     

Blocking the PCNA/NKp44 Checkpoint to Stimulate NK Cell Responses to Multiple Myeloma

Simple SummaryMembrane-associated PCNA is expressed on the surface of human MM cell lines and primary MM cells. Mab 14-25-9 interacts with membrane-associated PCNA and blocks its binding to NK-expressed NKp44, thus activating NK function. We showed that mAb 14-25-9 can serve as an immune checkpoint blocker, enhancing the function of NK cells on target human MM cell lines and primary cells.AbstractMultiple Myeloma (MM) is a devastating malignancy that evades immune destruction using multiple mechanisms. The NKp44 receptor interacts with PCNA (Proliferating Cell Nuclear Antigen) and may inhibit NK cells’ functions. Here we studied in vitro the expression and function of PCNA on MM cells. First, we show that PCNA is present on the cell membrane of five out of six MM cell lines, using novel anti-PCNA mAb developed to recognize membrane-associated PCNA. Next, we stained primary bone marrow (BM) mononuclear cells from MM patients and showed significant staining of membrane-associated PCNA in the fraction of CD38⁺CD138⁺ BM cells that contain the MM cells. Importantly, blocking of the membrane PCNA on MM cells enhanced the activity of NK cells, including IFN-γ-secretion and degranulation. Our results highlight the possible blocking of the NKp44-PCNA immune checkpoint by the mAb 14-25-9 antibody to enhance NK cell responses against MM, providing a novel treatment option.     

Influence of Y~(3+),Yb~(3+),Gd~(3+) cations on structural and electromagnetic properties of CuFe_2O_4 nanoferrites prepared via one step sol-gel method

Rare earths(REs) play a key role in distorting spinel structure by creating some defects at the lattice sites and make them suitable for magnetodielectric applications.In the present study,the nanoferrites of CuRE_(0.02)Fe_(1.98)O_4,where REs=Y~(3+),Yb~(3+),Gd~(3+),were prepared using one step sol-gel method.The prepared samples are copper ferrite(CFO),yttrium doped copper ferrite(Y-CFO),ytterbium doped copper ferrite(Yb-CFO) and gadolinium doped copper ferrite(Gd-CFO),respectively.The single-phase structure of all the REs doped nanoferrites was determined by X-ray diffraction(XRD) analysis.The porosity,agglomerations and grain size of the REs doped copper ferrite were examined using field emission scanning electron microscopy(FESEM) analysis.Fourier transform infrared spectroscopy(FTIR)elaborates the phase formation and environmental effects on the REs doped nanoparticles(NPs).The recorded room temperature M-H loops from a vibrating sample magnetometer(VSM) elucidate the magnetic properties of the REs doped spinel nanoferrites.The magnetic saturation(M_s) was calculated in the range of 23.08 to 51.78 emu/g.The calculated coercivity values(272.6 to 705.60 Oe) confirm the soft magnetic behavior of REs doped copper ferrites.Furthermore,the electromagnetic and dielectric properties were assessed using a Vector network analyzer(VNA) from 1 to 6 GHz.The permeability,permittivity,dielectric tangent loss and electric modulus of the REs doped spinel ferrites illustrate that the prepared NPs may be suitable for microwave and high frequency applications.     

Effect of aspect ratio on large deformation and necking of polyethylene

Objective of the study is to examine influence of aspect ratio of rectangular cross-section on the tri-axial stress state developed by necking in tensile specimens of polyethylene. The first part of the paper presents an experimental study that used two types of high-density polyethylene (HDPE) as sample material to identify the thickness-dependent relationship between engineering stress and elongation from tensile tests. The experimental study also shows that thinner specimens, i.e. higher aspect ratio, have lower neck propagation speed and higher flow stress, thus higher rate of energy consumption for the neck propagation. The second part of the paper presents finite element simulation of large deformation and necking in HDPE when subjected to uni-axial tension. True stress–strain relationship and governing equation for visco-plastic deformation are determined from the finite element simulation based on experimental data for the two HDPEs, which reveals influence of aspect ratio of cross-section on the stress state during the necking process. Results from the study indicate that plane-stress condition prevails when the aspect ratio increases, i.e. by decreasing the specimen thickness. The finite element simulation also supports the observation that necking in specimens with higher aspect ratio, i.e. thinner specimens, generates higher percentage of reduction in the thickness direction but lower in the width direction. The overall capability for the deformation endurance was found to improve by reducing the specimen thickness. The paper concludes that finite element simulation has successfully demonstrated the influence of aspect ratio of the cross-section on the stress state in the necking process. The paper also concludes that by combining experimental testing and finite element simulation, time-dependent deformation behaviour can be separated from the time-independent deformation behaviour, which is almost impossible to achieve based on the experimental techniques that rely purely on measurement only.     

Asymptotic stability, ℓ2 gain,boundness analysis,and control synthesis for switched systems: a switching frequency approach

In this paper, several concepts of switching frequency are introduced to analyze the properties and performance of switched systems in infinite as well as finite‐time intervals. The observation is very motivating that different system properties and performances depend on different switching frequencies. Sufficient conditions ensuring asymptotic stability, ?₂ gain performance, and state boundness are derived on the basis of the notions of switching frequency, respectively. Then, on the basis of the analysis results, the control synthesis problems are addressed. LMI‐based design algorithms are proposed to meet different control synthesis requirements. Numerical design examples are provided to demonstrate our results. Copyright © 2011 John Wiley & Sons, Ltd.     

New two CFOA-based sinusoidal RC oscillators with buffered outlet

Using a unified representation for a class of buffered-outlet two current-feedback operational amplifier (CFOAs)-based sinusoidal oscillators, new circuits of this type can be systematically discovered. A catalogue of four circuit structures, each structure realizing nine oscillator circuits, is presented. Moreover, using the RC:CR transformation, additional nine oscillator circuits can be obtained from each structure. While each circuit requires five passive elements, some of the circuits enjoy one or more of the following attractive features: use of grounded capacitors, feasibility of absorbing the parasitic components of the CFOAs and orthogonal tuning of the frequency and the startup condition of oscillation.     

Analysis of the decoupled uplink downlink technique for varying path loss exponent in multi-tier HetNet

In this paper, the impact of varying path loss exponent (PLE) on user association probability, decoupled uplink coverage probability as well as decoupled uplink average spectral efficiency in downlink uplink decoupled (DUDe) multi-tier heterogeneous networks, is investigated. We investigate the effect of the difference in path loss exponents in both macro and small cell environments over uplink network performance. It is assumed that the mobile user connected to the macro base station experience different path loss exponent as compared to when connected to small base station. It is observed that the difference of path loss exponents in both cases has significant effect on the user association probability, decoupled uplink coverage probability as well as decoupled uplink average spectral efficiency. Moreover, in order to further support key findings and make sound comparison between coupled and DUDe performance in varying PLE environment, generalized analytical expressions for coupled association probabilities, along with coupled uplink coverage probability and coupled uplink average spectral efficiency have been derived. The analytical results evaluated in this paper are compared with the computer simulation and found in good agreement. Our analysis shows that decoupling technique performs suboptimal for cases where the environments around macro and small base stations are different with respect to each other. The work explained in this paper highlights the limitation of applying DUDe technique in realistic conditions where the PLEs of cellular tiers are not exactly equal to one another.

     

Road Oriented Traffic Information System for Vehicular Ad hoc Networks

Over the last few years, vehicular ad hoc networks (VANETs) have gained popularity for their interesting applications. To make efficient routing decisions, VANET routing protocols require road traffic density information for which they use density estimation schemes. This paper presents a distributed mechanism for road vehicular density estimation that considers multiple road factors, such as road length and junctions. Extensive simulations are carried out to analyze the effectiveness of the proposed technique. Simulation results suggested that, the proposed technique is more accurate compared to the existing technique. Moreover, it facilitate VANET routing protocols to increase packet delivery ratio and reduce end-to-end delay.