Luciferase-transfected colon adenocarcinoma cell line (DLD-1) for use in Orthotopic Xenotransplantation studies

ABSTRACT: BACKGROUND: Renilla Luciferase reporter gene (rLuc) GL4.82 and GL4.13 promoter are key player in transfection, but precise knowledge of its targets in colon cancer remains limited. The aim of this study was to characterize the best transfection technique to produce a stable transfected colon DLD1 (colorectal adenocarcinoma cell line), therefore imaging based approaches were employed. RESULTS: DLD1 cells were transfected with a Plasmid (SV40-RLuc) carrying Renilla luciferase under the control of the SV-40 promoter, by using two different transfection techniques. Cells expressing the required DNA were isolated after antibiotic (Puramycin) selection. Clones of DLD-1/SV40-RLuc were produced using two different techniques (96 well plates and Petri dish) and their florescence intensity was recorded using IVIS machine (Calliper Life Sciences, Hopkinton, USA). Both techniques were characterized with the help of serial dilution technique. Results from this study substantiated that electroporation is the best. As expected, clones varied in their specific luciferase activity along with the dilutions. With the increase in cell concentration increase in intensity of florescence was recorded. CONCLUSIONS: Based on the results we are confident that this transfected cell line DLD-1/SV40-RLuc (colorectal adenocarcinoma cell line) is the best for further Orthotopic Xenotransplantation Studies and in-vivo experiments as well. Investigation shows that DLD1/SV-rLuc cells have gained little bit resistance against both drugs therefore further study is suggested to know the reasons.     

Microwave-promoted efficient conversion of acetophenones to 1,3,5-triarylbenzenes catalyzed by H3PW12O40 and nano-silica supported H3PW12O40 as reusable catalysts

H₃PW₁₂O₄₀ and nano-silica supported H₃PW₁₂O₄₀ were found to be efficient heterogeneous catalysts for the preparation of 1,3,5-triarylbenzenes via triple self condensation of acetophenones under microwave irradiation and solvent-free conditions. High yields, short reaction times, easy work-up, easy availability and handling, eco-friendly and reusability of the catalysts are the main aspects of the present method. The catalytic mechanism of 1,3,5-triarylbenzenes synthesis is also proposed.     

Performance improvement of spectral amplitude coding-optical code division multiple access systems using NAND detection with enhanced double weight code

The bit-error rate (BER) performance of the spectral amplitude coding-optical code division multiple access (SACOCDMA) system has been investigated by using NAND subtraction detection technique with enhanced double weight (EDW) code. The EDW code is the enhanced version of double weight (DW) code family where the code weight is any odd number and greater than one with ideal cross-correlation. In order to evaluate the performance of the system, we used mathematical analysis extensively along with the simulation experiment. The evaluation results obtained using the NAND subtraction detection technique was compared with those obtained using the complementary detection technique for the same number of active users. The comparison results revealed that the BER performance of the system using NAND subtraction detection technique has greatly been improved as compared to the complementary technique.     

Electrophoretic deposition of PVA coated hydroxyapatite on 316L stainless steel

Polyvinyl alcohol (PVA) coated hydroxyapatite was deposited onto a 316L stainless steel substrate by electrophoretic deposition. Deposition was carried out in a methanol suspension at pH 5.5 using a graphite rod as an anode. Parameters such as PVA concentration, deposition voltage and time were optimized to achieve a homogeneous, crack-free adhesive coating. Techniques such as X-ray diffraction and Fourier transform infrared spectroscopy were used to study the phase composition of the coated materials and the stability of hydroxyapatite in the presence of PVA.     

A useful scaffold based on acenaphthene exhibiting Cu2+ induced excimer fluorescence and sensing cyanide via Cu2+ displacement approach

A new simple organic scaffold based on acenaphthene 4 was designed and synthesized. The chromogenic and fluorogenic properties of 4 toward different metal ions and anions were investigated in H₂O/MeCN (8:2, v/v) solution. The probe 4 in the presence of Cu²⁺ exhibited strong static excimer emission at 507 nm along with a decrease in monomer emission at ～400 nm ratiometrically, attributed to a complexation through aldimine and amide groups of 4. Additionally, 4 upon interaction with different anions illustrated significant fluorescence enhancement with cyanide. However, interaction of complex, 4-Cu²⁺ with CN^? revealed fluorescence quenching attributed to formation of stable [Cu(CN)_x]^1?x species in the medium. A naked-eye sensitive fluorescent green color of solution was changed to blue. The mechanism of interaction between 4 and Cu²⁺ and sensing of cyanide through Cu²⁺ displacement approach was confirmed by the change in optical behaviors and ¹H NMR and ESI-MS spectral data analysis.     

The regularity of edge ideals of graphs

In this paper, we introduce some reduction processes on graphs which preserve the regularity of related edge ideals. Using these, an alternative proof for the theorem of R. Fröberg on the linearity of the resolutions of edge ideals is given.     

Dynamic characteristics of nanoindentation in Ni:A molecular dynamics simulation study

In this work,three-dimensional molecular dynamics simulation is carried out to elucidate the nanoindentation behaviour of single crystal Ni.The substrate indenter system is modelled using hybrid interatomic potentials including the manybody potential(embedded atom method) and two-body Morse potential.The spherical indenter is chosen,and the simulation is performed for different loading rates from 10 m/s to 200 m/s.Results show that the maximum indentation load and hardness of the system increase with the increase of velocity.The effect of indenter size on the nanoindentation response is also analysed.It is found that the maximum indentation load is higher for the large indenter whereas the hardness is higher for the smaller indenter.Dynamic nanoindentation is carried out to investigate the behaviour of Ni substrate to multiple loading-unloading cycles.It is observed from the results that the increase in the number of loading unloading cycles reduces the maximum load and hardness of the Ni substrate.This is attributed to the decrease in recovery force due to defects and dislocations produced after each indentation cycle.     

Modeling the interfacial effect on the yield strength and flow stress of thin metal films on substrates

It is shown in this paper that interfacial effects have a profound impact on the scale-dependent yield strength and strain hardening rates (flow stress) of metallic thin films on elastic substrates. This is achieved by developing a higher-order strain gradient plasticity theory based on the principle of virtual power and the laws of thermodynamics. This theory enforces microscopic boundary conditions at interfaces which relate a microtraction stress to the interfacial energy at the interface. It is shown that the film bulk length scale controls the size effect if a rigid interface is assumed whereas the interfacial length scale dominates if a compliant interface is assumed.     

Micromechanical theoretical and computational modeling of energy dissipation due to nonlinear vibration of hard ceramic coatings with microstructural recursive faults

Engine failures due to high-cycle fatigue during severe dynamic vibration have cost the US Air Force an estimated $400 million dollars per year over the past two decades. Therefore, structural materials that exhibit high damping capacities are desirable for mechanical vibration suppression and acoustic noise attenuation. Few experimental studies suggested that hard ceramic coatings, which are commonly used as thermal barrier coatings (TBCs) to protect engine components from high temperatures and corrosion, can also serve as passive dampers due to their unique microstructure which consists of several layers of splats with inter- and intra-microstructural recursive faults (micro-cracks). Therefore, the focus of this study is on the development of a fundamental understanding of the unique microstructural features and mechanisms responsible for this observed energy dissipation in ceramic coatings under nonlinear vibration through the development of a micromechanical computational framework. Inter- and intra-fatigue damage and internal friction is simulated through the development of thermodynamic-based nonlinear cohesive laws that consider interfacial degradation, debonding, plastic sliding, and Coulomb/contact friction between the interfaces of microstructural faults. Representative volume element-based micromechanical simulations are conducted in order to assess the main micromechanical mechanisms responsible for the experimentally observed nonlinear (amplitude- and frequency-dependent) damping in plasma sprayed hard ceramic coatings. It is concluded that the major part of energy dissipation is achieved through contact friction which results from sliding of the splat interfaces along the microstructural recursive faults. Energy dissipation due to progressive decohesion and evolution of new micro-cracks is not that significant as compared to energy dissipated due to increased friction from existing and new created faults. Therefore, internal friction is the main mechanism that makes TBCs effective dampers.