Generation of Helper Plasmids Encoding Mutant Adeno-associated Virus Type 2 Capsid Proteins with Increased Resistance against Proteasomal Degradation

Cardiolog

H∞ drive train control for hardware-in-the-loop simulation with a scaled dynamometer test bench

This paper presents the design and implementation of a robust drive train torque control for rotor inertia emulation in a dynamometer test bench application. The controller is part of a hardware-in-the-loop (HiL) framework for a 50 kW small test rig that enables a real-time interaction between test rig drive train and a rotor model. The control objective is to compensate undesired dynamics of the test rig, in order to guarantee emulation of model dynamics through HiL simulation within a desired bandwidth. Therefore, a robust H ∞ controller has been implemented taking advantage of the design in frequency domain. Furthermore, a Kalman filter has been incorporated providing the unmeasured variables in the presence of measurement noise. For the purpose of control performance evaluation, simulation results as well as experimental tests on the hardware are presented

Development of a Novel Thermally Stable Inhibitor Based on Furfuryl Alcohol for Mild Steel Corrosion in a 15% HCl Medium for Acidizing Application

Although corrosion inhibitors are widely used as a practical method for inhibiting metal corrosion in a variety of industries, their efficiency decreases dramatically at high temperatures. We present a thorough experimental and theoretical investigation into the inhibitory power of a novel thermally stable corrosion inhibitor based on furfuryl alcohol (FACI). The open-circuit potential, linear polarization resistance, potentiodynamic polarization, and electrochemical impedance spectroscopy techniques were used for the first time to evaluate FACI inhibition performance at 293, 313, 333, and 353 K. All electrochemical methods clearly indicated that FACI effectively suppressed corrosion of mild steel (MS) in the 15% HCl environment via a mixed-type mechanism at all temperatures investigated. The highest inhibition efficiencies of 95.4 and 90.8 at 333 and 353 K, respectively, were obtained in the solution containing 136 × 10-5 M FACI. FACI inhibited both the anodic and cathodic corrosion reactions of MS, but its main effect was on the cathodic hydrogen evolutions. The results of the Langmuir adsorption isotherm confirmed that the predominantly chemical adsorption of FACI molecules occurred on the MS surface. The experimental results were also validated using a combined DFT

Development of a Lentivirus Vector-Based Assay for Non-Destructive Monitoring of Cell Fusion Activity

Cardiolog

Comparison of LV.GS.GpLuc.v1- and LV.GS.GpLuc.v6-based cell fusion assay systems.

<p>(<b>A</b>): GpLuc production by proliferating or differentiating co-cultures of myoblasts^GS.GLuc or myoblasts^GS.GLuc+ with myoblasts-FLPe^NLS+ at different times after culture initiation. Cells were seeded in different ratios (<i>i.e.</i> 100∶0%, 90∶10% and 75∶25%). At 72 h after cell seeding the culture fluid was replaced by fresh culture medium with (growth conditions, no differentiation) or without (differentiation conditions) serum. Ninety-six h and 120 h later culture medium collected for luciferase activity measurement. Bars represent mean ± standard error of the mean (n = 3). (<b>B</b>): Fold change in luciferase activity calculated on the basis of the data presented in (A). For each experimental condition the average light production under growth conditions was the denominator and the mean of the RLUs produced under differentiation conditions was the numerator. RLUs, relative light units; G1, LV.GS.GpLuc.v1-based cell fusion assay; G6, LV.GS.GpLuc.v6-based cell fusion assay; MBs^GS.GLuc(+), myoblasts^GS.GLuc or myoblasts^GS.GLuc+; MBs-FLPe^NLS+, myoblasts-FLPe^NLS+.</p

Comparison of LV.GS.GpLuc.v1- and LV.GS.PpLuc-based cell fusion assay systems.

<p>(<b>A</b>): GpLuc and PpLuc production by proliferating or differentiating co-cultures of myoblasts^GS.GLuc or myoblasts^GS.PLuc with myoblasts-FLPe^NLS+ at different times after culture initiation. Cells were seeded in different ratios (<i>i.e.</i> 100∶0%, 90∶10%, 75∶25% and 50∶50%). At 72 h after cell seeding the culture fluid was replaced by fresh culture medium with (growth conditions, no differentiation) or without (differentiation conditions) serum. Ninety-six h and 120 h later samples (culture fluid for cultures containing myoblasts^GS.GLuc and cell lysates for cultures containing myoblasts^GS.PLuc) were harvested for luciferase activity measurements. Bars represent mean ± standard error of the mean (n = 3). (<b>B</b>): Fold change in luciferase activity calculated on the basis of the data presented in (A). For each experimental condition the average light production under growth conditions was the denominator and the mean of the RLUs produced under differentiation conditions was the numerator. RLUs, relative light units; G, LV.GS.GpLuc.v1-based cell fusion assay; P, LV.GS.PpLuc-based cell fusion assay; MBs^GS.Luc, myoblasts^GS.GLuc or myoblasts^GS.PLuc; MBs-FLPe^NLS+, myoblasts-FLPe^NLS+.</p

Analysis of FLPe^NLS+/− level, intracellular localization and enzymatic activity.

<p>(<b>A</b>): Western blotting analysis of whole protein lysates, nuclear cell fractions and cytosolic cell fractions of myoblasts-FLPe^NLS+ (+) and of myoblasts-FLPe^NLS− (−) maintained in growth medium (no differentiation) or exposed to differentiation conditions for 96 h (differentiation). (<b>B</b>): Luciferase activity measurements in culture media of myoblasts^GS.GLuc transduced with LV.FLPe^NLS+.PurR, LV.FLPe^NLS−.PurR or LV.PurR (negative control vector) representing different intervals (<i>i.e.</i> 0–24 h and 24–48 h) post transduction. Bars show mean ± standard error of the mean (n = 3). (<b>C</b>): Fold change in luciferase activity calculated on the basis of the data presented in (B). The average light production by samples of LV.PurR-transduced myoblasts^GS.GLuc was the denominator and the mean of the RLUs produced by LV.FLPe^NLS+.PurR-transduced myoblasts^GS.GLuc (NLS⁺) or by LV.FLPe^NLS−.PurR-transduced myoblasts^GS.GLuc (NLS⁻) was the numerator. NLS, nuclear localization signal; FLPe, molecularly evolved flippase; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; RLUs, relative light units.</p