Development of Anti-LRRC15 Small Fragments for Imaging Purposes Using a Phage-Display ScFv Approach

The human leucine-rich repeat-containing protein 15 (LRRC15) is a membrane protein identified as a marker of CAF (cancer-associated fibroblast) cells whose overexpression is positively correlated with cancer grade and outcome. Nuclear molecular imaging (i.e., SPECT and PET) to track LRRC15 expression could be very useful in guiding further therapeutic strategies. In this study, we developed an ScFv mouse phage-display library to obtain small fragment antibodies against human LRRC15 for molecular imaging purposes. Mice were immunized with recombinant human LRRC15 (hLRRC15), and lymph node cells were harvested for ScFv (single-chain variable fragment) phage-display analysis. The built library was used for panning on cell lines with constitutive or induced expression after transfection. The choice of best candidates was performed by screening various other cell lines, using flow cytometry. The selected candidates were reformatted into Cys-ScFv or Cys-diabody by addition of cysteine, and cloned in mammalian expression vectors to obtain batches of small fragments that were further used in site-specific radiolabeling tests. The obtained library was 1.2 × 10⁷ cfu/µg with an insertion rate >95%. The two panning rounds performed on cells permittedenrichment of 2 × 10⁻³. Screening with flow cytometry allowed us to identify 28 specific hLRRC15 candidates. Among these, two also recognized murine LRCC15 and were reformatted into Cys-ScFv and Cys-diabody. They were expressed transiently in a mammalian system to obtain 1.0 to 4.5 mg of Cys fragments ready for bioconjugation and radiolabeling. Thus, in this paper, we demonstrate the relevance of the phage-display ScFv library approach for the fast-track development of small antibodies for imaging and/or immunotherapy purposes.     

Development and Verification of Simulation Model Based on Real MANET Experiments for Transport Layer Protocols （UDP and TCP）

There is a lack of appropriate guidelines for realistic user traces, mobility models, routing protocols, considerations of real-life challenges, etc. for general-purpose mobile ad hoc networks (MANET). In this paper, four laptops are used in an open field environment in four scenarios to evaluate the performances of Internet control message protocol (ICMP) based ping and transmission control protocol (TCP) based streaming video applications using optimised link state routing (OLSR) implementation in an IEEE 802.11g wireless network. Corresponding simulations are developed in Network Simulator ns-2 by setting simulation parameters according to the real experiments. Difficulties faced to regenerate real-life scenarios have been discussed and the gaps between reality and simulation are identified. A setup guideline to produce realistic simulation results has been established.     

Silver-Decorated Silicon Nanostructures: Fabrication and Characterization of Nanoscale Terraces as an Efficient SERS-Active Substrate

Rich and highly dense surface-enhanced Raman (SERS) hotspots available in the SERS-active platform are highly anticipated in SERS measurements. In this work, conventional silicon wafer was treated to have wide exposure to terraces available within the silicon nanostructures (Si-NSs). High-resolution field emission scanning electron microscopic (FESEM) investigations confirmed that the terraces were several microns wide and spread over different steps. These terraces were further decorated with silver nanoparticles (Ag-NPs) of different shapes and sizes to achieve SERS-active hotspots. Based on more than 150 events, a histogram of the size distribution of Ag-NPs indicated a relatively narrow size distribution, 29.64 ± 4.66 nm. The coverage density was estimated to be ~4 × 10¹⁰ cm⁻². The SERS-activity of Ag-NPs -decorated Si-NSs was found to be enhanced with reference to those obtained in pristine Si-NSs. Finite difference time domain models were developed to support experimental observations in view of electromagnetic (EM) near-field distributions. Three archetype models; (i) dimer of same constituent Ag-NPs, (ii) dimer of different constituent Ag-NPs, and (iii) linear trimer of different constituent Ag-NPs were developed. EM near-field distributions were extracted at different incident polarizations. Si-NSs are well-known to facilitate light confinement, and such confinement can be cascaded within different Ag-NPs-decorated terraces of Si-NSs.     

Dielectric relaxation in complex perovskite Ba(Bi1/2Ta1/2)O3

A new lead-free perovskite Ba(Bi_1/2Ta_1/2)O₃ was prepared by conventional ceramic fabrication technique at 1,200 °C/4 h in air atmosphere. The crystal symmetry, space group and unit cell dimensions were determined from the experimental results using FullProf software. The crystallite size and lattice strain were estimated from Williamson-Hall approach. XRD analysis of the compound indicated the formation of a single-phase cubic structure with the space group Pm3m. EDAX and SEM studies were carried out in order to evaluate the quality and purity of the compound. Permittivity data showed a low temperature coefficient of capacitance (T _CC  < 4%) up to +125 °C. Complex impedance analyses suggested the dielectric relaxation to be of non-Debye type. Electric modulus studies supported the hopping type of conduction in Ba(Bi_1/2Ta_1/2)O₃.     

Properties of PET/clay nanocomposite films

Polyethylene terephthalate (PET) nanocomposite films were prepared by cast extrusion followed by uniaxial stretching, using chill rolls. Transmission electron microscopy (TEM) and wide angle X‐ray diffraction (WAXD) showed that the clay layers were aligned in the machine direction (MD) in the PET/clay nanocomposite (PCN) films. Differential scanning calorimetry (DSC) showed that PCN films have higher crystallinity than the neat PET films, possibly due to the nucleating role of the silicate layers. The PCN films became hazier as the clay content increased, but the film transparency remained in the acceptable range. Oxygen permeability of the PCN films decreased by 23% compared to the neat PET film. This is comparable with predictions of models proposed in the literature. Silicate incorporation brought about 20% increase in the tensile modulus, while the puncture and tear propagation resistance were reduced, due to brittleness of the PCN films. The measured modulus (1.7 GPa) was somewhat smaller than the values predicted using the Pseudoinclusion model (2.1 GPa). POLYM. ENG. SCI., 2012. © 2011 Society of Plastics Engineers     

Investigation on the effect of spinning conditions on the properties of hollow fiber membrane for hemodialysis application

Polyethersulfone (PES) hollow fiber membranes were fabricated via the dry‐wet phase inversion spinning technique, aiming to produce an asymmetric, micro porous ultrafiltration hollow‐fiber specifically for hemodialysis membrane. The objective of this study is to investigate the effect of spinning conditions on the morphological and permeation properties of the fabricated membrane. Among the parameters that were studied in this work are air gap distance, dope extrusion rate, bore fluid flow rate, and the take‐up speed. The contact angle was measured to determine the hydrophilicity of the fibers. Membrane with sufficient hydrophilicity properties is desired for hemodialysis application to avoid fouling and increase its biocompatibility. The influences of the hollow fiber's morphology (i.e., diameter and wall thickness) on the performance of the membranes were evaluated by pure water flux and BSA rejection. The experimental results showed that the dope extrusion rate to bore fluid flow rate ratio should be maintained at 1:1 ratio to produce a perfectly rounded asymmetric hollow fiber membrane. Moreover, the flux of the hollow fiber spun at higher air gap distance had better flux than the one spun at lower air gap distance. Furthermore, spinning asymmetric hollow fiber membranes at high air gap distance helps to produce a thin and porous skin layer, leading to a better flux but a relatively low percentage of rejection for BSA separation. Findings from this study would serve as primary data which will be a useful guide for fabricating a high performance hemodialysis hollow fiber membrane. © 2016 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2016 , 133, 43633.     

Synthesis,Characterization and Surface Properties of Amidosulfobetaine Surfactants Bearing Odd-Number Hydrophobic Tail

Three amidosulfobetaine surfactants were synthesized namely: 3-(N-pentadecanamidopropyl-N,N-dimethyl ammonium) propanesulfonate (2a); 3-(N-heptadecanamidopropyl-N,N-dimethyl ammonium) propanesulfonate (2b), and 3-(N-nonadecanamidopropyl-N,N-dimethyl ammonium) propanesulfonate (2c). These surfactants were prepared by direct amidation of commercially available fatty acids with 3-(dimethylamino)-1-propylamine and subsequent reaction with 1,3-propanesultone to obtain quaternary ammonium salts. The synthesized surfactants were characterized by IR, NMR and mass spectrometry. Thermogravimetric analysis (TGA) results showed that the synthesized surfactants have excellent thermal stability with no major thermal degradation below 300 °C. The critical micelle concentration (CMC) values of the surfactants 2a and 2b were found to be 2.2 × 10^?4 and 1.04 × 10^?4 mol/L, and the corresponding surface tension (γ_CMC) values were 33.14 and 34.89 mN m^?1, respectively. The surfactants exhibit excellent surface properties, which are comparable with conventional surfactants. The intrinsic viscosity of surfactant (2b) was studied at various temperatures and concentrations of multi-component brine solution. The plot of natural logarithm of relative viscosity versus surfactant concentration obtained from Higiro et al. model best fit the surfactant behavior. Due to good salt resistance, excellent surface properties and thermal stability, the synthesized surfactant has potential to be used in various oil field applications such as enhanced oil recovery, fracturing, acid diversion, and well stimulation.     

A comparative study of thermal and hydraulic autofrettage

Thermal autofrettage is a potential process to generate beneficial compressive residual stresses at and around the inner wall of a thickwalled cylinder for increasing its pressure carrying capacity. Due to its simplicity and inexpensive arrangement, it can compete with the conventional hydraulic autofrettage process. In this work, a comparative study of the thermal autofrettage and the hydraulic autofrettage is carried out. As the thermal autofrettage does not require hydraulic power pack, the process is more economical than the hydraulic autofrettage. The thermal autofrettage process is also studied for the thick-walled cylindrical vessels subjected to high thermal gradient with or without pressure and is compared with the hydraulic autofrettage. Comparison shows that for cylinders subjected to high thermal gradient without pressure, the thermal autofrettage is superior to the hydraulic autofrettage.