Rapid discovery of potent siRNA-containing lipid nanoparticles enabled by controlled microfluidic formulation

The discovery of potent new materials for in vivo delivery of nucleic acids depends upon successful formulation of the active molecules into a dosage form suitable for the physiological environment. Because of the inefficiencies of current formulation methods, materials are usually first evaluated for in vitro delivery efficacy as simple ionic complexes with the nucleic acids (lipoplexes). The predictive value of such assays, however, has never been systematically studied. Here, for the first time, by developing a microfluidic method that allowed the rapid preparation of high-quality siRNA-containing lipid nanoparticles (LNPs) for a large number of materials, we have shown that gene silencing assays employing lipoplexes result in a high rate of false negatives (~90%) that can largely be avoided through formulation. Seven novel materials with in vivo gene silencing potencies of >90% at a dose of 1.0 mg/kg in mice were discovered. This method will facilitate the discovery of next-generation reagents for LNP-mediated nucleic acid delivery.     

A novel sensor of cysteine self-assembled monolayers over gold nanoparticles for the selective determination of epinephrine in presence of sodium dodecyl sulfate

A novel sensor of cysteine self-assembled monolayers over gold nanoparticles modified gold electrode has been constructed for the determination of epinephrine in presence of sodium dodecyl sulfate (Au/Au(nano)-CysSDS). Electrochemical investigation and characterization of the modified electrode are achieved using cyclic voltammetry, linear sweep voltammetry, and scanning electron microscopy. The Au/Au(nano)-CysSDS electrode current signal is remarkably stable via repeated cycles and long term stability, due to the strong Au-S bond, compared to the Au/Au(nano) electrode. The catalytic oxidation peak currents obtained from linear sweep voltammetry (LSV) increased linearly with increasing epinephrine concentrations in the range of 2 to 30 μmol L(-1) and 35 to 200 μmol L(-1) with correlation coefficients of 0.9981 and 0.9999 and a limit of detection of 0.294 nmol L(-1) and 1.49 nmol L(-1), respectively. The results showed that Au/Au(nano)-CysSDS can selectively determine epinephrine in the coexistence of a large amount of uric acid and glucose. In addition, a highly selective and simultaneous determination of tertiary mixture of ascorbic acid, epinephrine, and acetaminophen is explored at this modified electrode. Excellent recovery results were obtained for determination of epinephrine in spiked urine samples at the modified electrode. Au/Au(nano)-CysSDS can be used as a sensor with excellent reproducibility, sensitivity, and long term stability.     

In vitro selection and characterization of deoxyribonucleic acid aptamers for digoxin

The low therapeutic index of digoxin necessitates careful monitoring of its serum levels. Most of digoxin immunoassays suffer from interferences with digoxin-like immunoreactive substances. Since aptamers have been shown to be highly specific for their targets, the aim of this study was to develop DNA aptamers for this widely used cardiac glycoside. Digoxin was coated onto the surface of streptavidin magnetic beads. DNA aptamers against digoxin were designed using Systematic Evolution of Ligands by Exponential enrichment method (SELEX) by 11 iterative rounds of incubation of digoxin-coated streptavidin magnetic beads with synthetic DNA library, DNA elution, electrophoresis and PCR amplification. The PCR product was cloned and sequenced. Binding affinity was determined using digoxin–BSA conjugate, coated onto ELISA plate. Inhibitory effect of anti-digoxin aptamer was conducted using isolated guinea-pig atrium. Three aptamers (D1, D2 and D3) were identified. Binding studies of fluorescein-labeled truncated (without primer binding region) D1 and D2 and full length D1 anti-digoxin aptamers were performed and their corresponding dissociation constants values were 8.2 × 10⁻⁹, 44.0 × 10⁻⁹ and 17.8 × 10⁻⁹ M, respectively. This is comparable to what other workers have obtained for interaction of monoclonal antibodies raised against digoxin. There was little difference in binding affinity between full length and truncated anti-digoxin D1 aptamer. D1 anti-digoxin aptamer also inhibited the effects of digoxin on the isolated guinea-pig atrium. D1 anti-digoxin aptamer distinguished between digoxin and ouabain in both tissue study and binding experiments. Our finding indicated that D1 anti-digoxin aptamer can selectively bind to digoxin. Further studies might show its suitability for use in digoxin assays and as a therapeutic agent in life-threatening digoxin toxicity.     

Optimization strategies for radiation induced grafting of 4-vinylpyridine onto poly(ethylene-co-tetraflouroethene) film using Box-Behnken design

The radiation induced grafting of 4-vinylpyridine (4-VP) onto poly(ethylene-co-tetrafluoroethene) (ETFE) was optimized using the Box-Behnken factorial design available in the response surface method (RSM). The optimized grafting parameters; absorbed dose, monomer concentration, grafting time and reaction temperature were varied in four levels to quantify their effect on the grafting yield (GY). The validity of the statistical model was supported by the small deviation between the predicted (GY=61%) and experimental (GY=57%) values. The optimum conditions for enhancing GY were determined at the following values: monomer concentration of 48 vol%, absorbed dose of 64 kGy, reaction time of 4 h and temperature of 68 °C. A comparison was made between the optimization model developed for the present grafting system and that for grafting of 1-vinylimidazole (1-VIm) onto ETFE to confirm the validly and reliability of the Box-Behnken for the optimization of various radiation induced grafting reactions. Fourier transform infrared (FTIR), thermogravimetric analysis (TGA) and X-ray diffraction (XRD) were used to investigate the properties of the obtained films and provide evidence for grafting.     

UV photodetector behavior of 2D ZnO plates prepared by electrochemical deposition

Zinc oxide (ZnO) with 2D hexagonal structure was successfully prepared using electrochemical deposition (ECD) method on a quartz substrate pre-coated with indium-doped ZnO layer. The X-ray diffraction of the prepared sample confirmed that the hexagonal structure had a dominant c-axis orientation. The scanning electron microscopy revealed the 2D hexagonal structure of the prepared ZnO.     

Preparation of chemically deposited thin films of CdS/PbS solar cell

The present paper reports the preparation of a solar cell which has a cross-sectional scheme: ITO/CdS/PbS, containing a commercially transparent conductive ITO; chemically deposited n-type CdS (340 nm) and absorbed layer of p-type PbS (1400 nm). The structural and optical properties of the constituent films are presented. X-ray diffraction showed that all of the thin films are polycrystalline. Using scanning electron microscopy, the present study revealed that the films have uniform surface morphology over the substrate. The solar cell was characterized by determining the open circuit voltage, short-circuit current density, and J–V under 40 mW/cm² solar radiation. The efficiency of the solar cells was 1.35%, which is much higher (0.041, 0.5 and 0.1–0.4%) and slightly smaller (1.65%) than some solar cells reported in the literature.     

D3→FJ emission of Tb doped sol–gel silica

Amorphous silica samples doped with 0.1 and 1 mol% of terbium (Tb) were synthesized by the sol–gel method. In addition to the green light associated with ⁵D₄→⁷F_J transitions of Tb³⁺, the sample containing 0.1 mol% also emitted blue light as a result of ⁵D₃→⁷F_J transitions during photoluminescence (PL) measurements. As a result of concentration quenching this blue emission was not observed for the samples doped with the higher concentration (1 mol%). However the blue ⁵D₃ →⁷F_J emission was observed in the 1 mol% doped samples during cathodoluminescence (CL) measurements. Since a rough calculation indicated that the excitation rate in the CL system where the blue emission is observed may be similar to a laser PL system under conditions where the blue emission is not observed, the difference is attributed to the nature of the excitation sources. It is suggested that during the CL excitation incident electrons can reduce non-luminescent Tb⁴⁺ ions in the silica, substituting for Si⁴⁺ ions, to the excited (Tb³⁺)? state and that these are responsible for the blue emission, which does not occur during PL excitation.