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.     

Deposition thickness based high-throughput nano-imprint template

International Technology Roadmap for Semiconductors 2003 projected nano-imprint lithography has the potential of high throughput, sub-20 nm resolution, and low cost [S.Y. Chou, P.R. Krauss, P.J. Renstrom, Appl. Phys. Lett. 67 (1995) 3144; Science 272 (1996) 85, J.A. Rogers, C. Mirkin, Mater. Res. Bull. 26 (2001)]. For nano-imprint lithography, a template with 1X resolution is required. The existing industrial infrastructure for supporting deep ultra violet 4X photo masks by e-beam and/or a laser beam scanning writer does not offer pitch (center-to-center distance of an array of patterned lines) less than ∼60 nm [<http://public.itrs.net/2003ITRS>]. For nano-imprint lithography to be accepted across the industry, a reproducible simple fabrication process to make a high resolution, single emboss template is essential [L. Jay Guo, J. Phys. D: Appl. Phys. 37 (2004) R123-R141]. Here we show, a general fabrication method and fabricated nano-imprint templates with sub-15 nm template line width and 10 nm pitch length through out the entire 200 mm wafer, varying the deposition thickness of multiple alternate films, using atomic layer deposition. Although multilayer nano-imprint templates and their exciting use have been demonstrated, [W.J. Dauksher et al., J. Vac. Sci. Technol. B 22 (2004) 3306, B. Heidari, et al., The 49th international conference on electron, ion and photon beam technology and nanofabrication, Orlando, Florida, 2005, William M. Tong, et al., Proc. SPIE 5751 (2005) 46-55, N.A. Melosh, A. Boukai, F. Diana, B. Gerardot, A. Badolato, P.M. Petroff, J.R. Heath, Science 300 (2003) 112] such a small pitch was not shown and either complex lattice mismatch-based epitaxially grown films or unconventional etch chemistry was used. The bare necessity was a simple and economical fabrication process for a high throughput nano-imprint template. In that context, we have developed a template fabrication process using classical micro-fabrication techniques. Successful use of these techniques made the template fabrication process simple, economical, and expedient. Also a novel technique to provide flexible and accurate alignment for nanowire patterning has been described. In this technique, nanowire patterning is accomplished on the entire wafer with a single impression. Industry level batch-fabrication of our scheme illustrates its reproducibility and manufacturability. We anticipate, this simple, economical and time saving technique will help researchers and developers to perform their experiment on nano-scale feature patterned substrates easily and conveniently.     

A pseudojoint estimation of time delay and scale factor for M-wave analysis

A pseudojoint estimation of time scale and time delay between an unknown deterministic transient type signal and a reference signal is proposed. The method is based on the separation between the estimations of the two dependent parameters. The time autocorrelation function (TACF) preserves the time scale and is invariant with respect to the time delay between the signals. The time scale factor can, thus, be estimated independently from time delay using the TACFs of the two signals. After estimating the time scale factor, the signal can be scaled by the estimated amount. The time delay is then estimated without bias due to the time scale factor. To obtain high resolution joint estimates, the time scale factor is estimated in the scale domain from the scale transforms of the TACFs of the two signals. The proposed method has low computational cost. Moreover, the results on synthetic signals show good performance of the method with respect to the Cramér-Rao Lower Bound and the joint Maximum Likelihood Estimation. A possible application of the technique to the analysis of electromyogram (EMG) signals detected during electrically elicited contractions is also presented. In a few representative cases, it is shown that the time scale estimate reveals myoelectric manifestations of muscle fatigue and is less affected by M-wave truncation than spectral EMG attributes.     

GWO‐C: Grey wolf optimizer‐based clustering scheme for WSNs

A wireless sensor network (WSN) is a prominent technology that could assist in the fourth industrial revolution. Sensor nodes present in the WSNs are functioned by a battery. It is impossible to recharge or replace the battery, hence energy is the most important resource of WSNs. Many techniques have been devised and used over the years to conserve this scarce resource of WSNs. Clustering has turned out to be one of the most efficient methods for this purpose. This paper intends to propose an efficient technique for election of cluster heads in WSNs to increase the network lifespan. For the achievement of this task, grey wolf optimizer (GWO) has been employed. In this paper, the general GWO has been modified to cater to the specific purpose of cluster head selection in WSNs. The objective function for the proposed formulation considers average intra‐cluster distance, sink distance, residual energy, and CH balancing factor. The simulations are carried out in diverse conditions. On comparison of the proposed protocol, ie, GWO‐C protocol with some well‐known clustering protocols, the obtained results prove that the proposed protocol outperforms with respect to the consumption of the energy, throughput, and the lifespan of the network. The proposed protocol forms energy‐efficient and scalable clusters.     

Queue length‐based load balancing in data center networks

Traffic load balancing in data centers is an important requirement. Traffic dynamics and possibilities of changes in the topology (e.g., failures and asymmetries) make load balancing a challenging task. Existing end‐host–based schemes either employ the predominantly used ECN or combine it with RTT to get congestion information of paths. Both congestion signals, ECN and RTT, have limitations; ECN only tells whether the queue length is above or below a threshold value but does not inform about the extent of congestion; similarly, RTT in data center networks is on the scale of up to few hundreds of microseconds, and current data center operating systems lack fine‐grained microsecond‐level timers. Therefore, there is a need of a new congestion signal which should give accurate information of congestion along the path. Furthermore, in end‐host–based schemes, detecting asymmetries in the topology is challenging due to the inability to accurately measure RTT on the scale of microseconds. This paper presents QLLB, an end‐host–based, queue length–based load balancing scheme. QLLB employs a new queue length–based congestion signal that gives an exact measure of congestion along the paths. Furthermore, QLLB uses relative‐RTT to detect asymmetries in the topology. QLLB is implemented in ns‐3 and compared with ECMP, CONGA, and Hermes. The results show that QLLB significantly improves performance of short flows over the other schemes and performs within acceptable level, of CONGA and Hermes, for long flows. In addition, QLLB effectively detects asymmetric paths and performs better than Hermes under high loads.     

A Distributed and Elastic Application Processing Model for Mobile Cloud Computing

The latest developments in mobile computing technology have increased the computing capabilities of smart mobile devices (SMDs). However, SMDs are still constrained by low bandwidth, processing potential, storage capacity, and battery lifetime. To overcome these problems, the rich resources and powerful computational cloud is tapped for enabling intensive applications on SMDs. In Mobile Cloud Computing (MCC), application processing services of computational clouds are leveraged for alleviating resource limitations in SMDs. The particular deficiency of distributed architecture and runtime partitioning of the elastic mobile application are the challenging aspects of current offloading models. To address these issues of traditional models for computational offloading in MCC, this paper proposes a novel distributed and elastic applications processing (DEAP) model for intensive applications in MCC. We present an analytical model to evaluate the proposed DEAP model, and test a prototype application in the real MCC environment to demonstrate the usefulness of DEAP model. Computational offloading using the DEAP model minimizes resources utilization on SMD in the distributed processing of intensive mobile applications. Evaluation indicates a reduction of 74.6% in the overhead of runtime application partitioning and a 66.6% reduction in the CPU utilization for the execution of the application on SMD.