Computationally Guided Monomerization of Red Fluorescent Proteins of the Class Anthozoa

Red fluorescent proteins (RFPs) have attracted significant engineering focus because of the promise of near infrared fluorescent proteins, whose light penetrates biological tissue, and which would allow imaging inside of vertebrate animals. The RFP landscape, which numbers ~200 members, is mostly populated by engineered variants of four native RFPs, leaving the vast majority of native RFP biodiversity untouched. This is largely due to the fact that native RFPs are obligate tetramers, limiting their usefulness as fusion proteins. Monomerization has imposed critical costs on these evolved tetramers, however, as it has invariably led to loss of brightness, and often to many other adverse effects on the fluorescent properties of the derived monomeric variants. Here we have attempted to understand why monomerization has taken such a large toll on Anthozoa class RFPs, and to outline a clear strategy for their monomerization. We begin with a structural study of the far-red fluorescence of AQ143, one of the furthest red emitting RFPs. We then try to separate the problem of stable and bright fluorescence from the design of a soluble monomeric β-barrel surface by engineering a hybrid protein (DsRmCh) with an oligomeric parent that had been previously monomerized, DsRed, and a pre-stabilized monomeric core from mCherry. This allows us to use computational design to successfully design a stable, soluble, fluorescent monomer. Next we took HcRed, which is a previously unmonomerized RFP that has far-red fluorescence (λemission = 633 nm) and attempted to monomerize it making use of lessons learned from DsRmCh. We engineered two monomeric proteins by pre-stabilizing HcRed’s core, then monomerizing in stages, making use of computational design and directed evolution techniques such as error-prone mutagenesis and DNA shuffling. We call these proteins mGinger0.1 (λem = 637 nm / Φ = 0.02) and mGinger0.2 (λem = 631 nm Φ = 0.04). They are the furthest red first generation monomeric RFPs ever developed, are significantly thermostabilized, and add diversity to a small field of far-red monomeric FPs. We anticipate that the techniques we describe will be facilitate future RFP monomerization, and that further core optimization of the mGingers may allow significant improvements in brightness.

Control of polarization signal distortion by frequency domain phase conjugation in optical fiber systems

Optical frequency domain phase conjugation (FDPC) is based on phase conjugation of spectrum of an input signal. It is equivalent to the phase conjugation and the time reversal of the temporal envelope of an input signal. The use of FDPC to control polarization signal distortion in birefringent optical fiber systems is proposed. Evolution of polarization signals in the system using midway FDPC is analyzed theoretically and simulated numerically. It is shown that the distortion of polarization signals can be controlled effectively by FDPC. The impairments due to dispersion and nonlinear effects can be suppressed simultaneously.

Influence of voltage and frequency dimming on power losses in HF electronic ballasts for compact fluorescent lamps

Compact Fluorescent Lamps (CFL) incorporating electronic ballasts are widely used in lighting. In many cases the ability to dim the lamp is a requirement Dimming can be achieved by varying the voltage supplied to the inverter or by changing the switching frequency of the inverter. The effect of dimming by both approaches on the power losses in the inverter is studied in this work. The lamp and associated inverter has been modeled in PSPICE, using a behavioral model for the CFL. Predicted losses are in good agreement with experimental data obtained from calorimetry. The model was then used to determine the distribution of losses within the inverter, enabling a comparison of the effects of the two dimming methods to be made. © 2006 IEEE.

Losses in inductor-less piezoelectric transformer ballast for compact fluorescent lamps

In this study an inductor-less piezoelectric transformer (PT) based ballast for a 5 W CFL has been designed and simulated. The predictions of circuit currents and losses closely match experimentally measured values. The total simulated loss figure was confirmed against practically determined losses using a precision mini-calorimeter. Using simulation to disaggregate the total loss figure, it is seen that the PT makes the largest contribution to the total losses in such ballast.

Losses in self-oscillating ballasts for electrode-type compact fluorescent lamps

Commercially available integrated compact fluorescent lamps (CFLs) use self-resonant ballasts on grounds of simplicity and cost. To understand how to improve ballast efficiency, it is necessary to quantify the losses. The losses occurring in these ballasts have been directly measured using a precision mini-calorimeter. In addition, a Pspice model has been used to simulate the performance of an 18 W integrated CFL. The lamp has been represented by a behavioural model and Jiles-Atherton equations were used to model the current transformer core. The total loss is in close agreement with measurements from the mini-calorimeter, confirming the accuracy of the model. The total loss was then disaggregated into component losses by simulation, showing that the output inductor is the primary source of loss, followed by the inverter switches. © 2011 The Institution of Engineering and Technology.

Perylene-based fluorescent liquid crystal dye guest-host mixtures

We present a study on a series of dye guest-host mixtures using fluorescent perylene-based molecules as the guest dye in an organosiloxane host. These hosts have temperature-independent switching, at room temperature, through 90° for fields of the order of 10 Vrms/μm. Perylene molecules have been grafted onto the organosiloxane moiety via an alkyl spacer producing novel and rugged fluorescent dyes that are readily miscible in the host. Micro-separation of the low molar mass siloxane groups in the mesophases tend to form smectic phases. These planes produce an effective two-dimensional polymer backbonethat engenders the rugged mechanical properties of polymeric liquid crystals onto these low molar mass ferroelectric liquid crystals. In this study we show how the introduction of the dye molecules affects the electro-optic properties of the organosiloxane host. © 2001 OPA (Overseas Publishers Association) N.V. Published by license under the Gordon and Breach Science Publishers imprint, a member of the Taylor & Francis Group,.