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.

连续工作的体布拉格光栅外腔半导体激光器的温度特性

对体布拉格光栅(VBG)作为波长选择元件的外腔半导体激光器的波长锁定进行了实验研究,报道了连续运转输出功率达43.5 W的半导体激光器阵列的体布拉格光栅波长锁定实验结果,给出了不同热沉温度下的稳定的波长锁定结果,说明采用体布拉格光栅外腔将减小半导体激光器的温控压力。实验中发现,随着注入电流的增大,输出激光功率逐渐增强,锁定的激射波长向长波长方向偏移。在输出功率为34.5 W时,波长红移约0.56 nm。这一移动与实验测量的体布拉格光栅的温度特性相吻合。连续和高占空比运行、高输出功率情况下,在器件的设计和使用时应该考虑这一效应。

Ce：Mn：LiNbO3晶体紫外记录研究

采用紫外光作记录光在Ce：Mn：LiNbO3晶体中实现非挥发全息记录，灵敏度可达0．0803cm／J，衍射效率（固定）为5．07％，比采用红光为记录光，紫外光为敏化光的非挥发双中心记录方案均提高了50多倍。分析表明，采用紫外光作为记录光，深能级电子被激发比例极大提高，参与光折变过程的电子平均运动周期变短．提高了衍射效率和灵敏度；深浅能级电子光栅的同相位，使得固定空间电荷场变强。文中还研究了退火对记录性能的影响。

Mechanism of red colouration in Tovel-See (”Conclusions” only) [Translation from: Archiv fur Hydrobiologie 38 299-302, 1941]

Cases of red colouration in small lake basins, due to the abundant appearance of microorganisms have long been known. Usually it is caused by a fast, sudden, intensive propagation (so called ”bloom”) of Cyanophycae and bacteria. (e.g. Oscillatoracae, thiobacteria etc.). An exception to this is the red colouration of Tovel-See, an alpine lake basin in the Dolomites of the Brenta group (Trentino), lying at a height of 1178 m and hidden in the woodland of a valley. Here the red bloom has a double rhythm: a daily and a yearly rhythm. The colouration of one part of the lake takes place in the warmest months of the year (i.e. July, August, September) and in the middle hours of the day. The immediate origin of the bloom has been known for a long time: it is caused by the Peridinacae Glenodinium sanguineum. This paper describes the phenomenon of red colouration of the lake and discusses its conditions.

Nonvolatile holograms in LiNbO3 : Fe : Cu by use of the bleaching effect

We report our observation of a bleaching effect under an ultraviolet exposure in LiNbO3:Fe:Cu crystals. Two three-step recording-transferring-fixing schemes are proposed to record nonvolatile photorefractive holograms in such crystals. In the schemes two red laser beams and an ultraviolet illumination are used selectively to write the charge grating in the shallow-level Fe centers, to develop the charge grating in the deep-level Cu centers by transferring the charge grating in the Fe centers, and to fix only the charge grating in the Cu centers for unerasable read-out. Experimental results, verifications, and an optimal recording scheme are given. A comparison of the lithium niobate crystals of the same double-doping system of Fe:Mn, Ce:Mn, Ce:Cu, and Fe:Cu is outlined. (C) 2002 Optical Society of America.

Prescription for optimizing holograms in LiNbO3 : Fe : Mn

The photorefractive holographic dynamics of grating formation in photochromic doubly doped LiNbO3:Fe:Mn crystal is studied numerically and analytically in terms of the two-center model of Kukhtarev Et al. [Ferroelectrics 22, 949 (1979)]. The relations among the recorded and fixed space-charge fields and the doping densities, the oxidation-reduction states of the fields, and the intensities of UV-sensitizing and red recording beams are studied. Important conditions and effects are feued, and an optimal prescription for material doping and oxidation-reduction processing is suggested in which the crystal can be strongly oxidized and the Mn-doping density is smaller than the Fe-doping density. (C) 2000 Optical Society of America. OCIS codes: 050.7330, 190.5330, 090.2900.

Nonvolatile photorefractive holograms in LiNbO3 : Cu : Ce crystals

We report experimental and theoretical studies of nonvolatile photorefractive holographic recording in LiNbO3:Cu:Ce crystals with two illumination schemes: (1) UV light for sensitization and a red interfering pattern for recording and (2) blue light for sensitization and a red pattern for recording. The results show that the oxidized LiNbO3:Cu:Ce crystals can provide high, persistent refractive-index modulation with weak lightinduced scattering. The optimal working conditions and the prescription for doping and oxidation-reduction processing that yields the maximum refractive-index modulation are discussed. (C) 2000 Optical Society of America OCIS codes: 050.7330, 190.5330, 090.2900.

Frequency upconversion properties of Er3+/Yb3+-codoped lead-germanium-bismuth oxide glasses

The frequency upconversion properties of Er3+/Yb3+-codoped heavy metal oxide lead-germanium-bismuth oxide glasses under 975 mn excitation are investigated. Intense green and red emission bands centered at 536, 556 and 672 run, corresponding to the H-2(1/2) --> I-4(15/2), S-4(3/2) --> I-4(15/2) and F-4(9/2) -->I-4(15/2) transitions of Er3+, respectively, were simultaneously observed at room temperature. The influences of PbO on upconversion intensity for the green (536 and 556 nm) and red (672 nm) emissions were compared and discussed. The optimized rare earth doping ratio of Er3+ and Yb3+, is 1:5 for these glasses, which results in the stronger upconversion fluorescence intensities. The dependence of intensities of upconversion emission on excitation power and possible upconversion mechanisms were evaluated and analyzed. The structure of glass has been investigated by means of infrared (IR) spectral analysis. The results indicate that the Er3+/Yb3+-codoped heavy metal oxide lead-germanium-bismuth oxide glasses may be a potential materials for developing upconversion fiber optic devices. (C) 2006 Published by Elsevier Ltd.

Absorption characteristic and nonvolatile holographic recording in LiNbO3 : Cr : Cu crystals

The absorption characteristic of lithium niobate crystals doped with chromium and copper (Cr and Cu) is investigated. We find that there are two apparent absorption bands for LiNbO3:Cr:Cu crystal doped with 0.14 wt.% Cr2O3 and 0.011 wt.% CuO; one is around 480 nm, and the other is around 660 nm. With a decrease in the doping composition of Cr and an increase in the doping composition of Cu, no apparent absorption band in the shorter wavelength range exists. The higher the doping level of Cr, the larger the absorbance around 660 nm. Although a 633 nm red light is located in the absorption band around 660 nm, the absorption at 633 nm does not help the photorefractive process; i.e., unlike other doubly doped crystals, for example, LiNbO3:Fe:Mn crystal, a nonvolatile holographic recording can be realized by a 633 nm red light as the recording light and a 390 nm UV light as the sensitizing light. For LiNbO3:Cr:Cu crystals, by changing the recording light from a 633 nm red light to a 514 nm green light, sensitizing with a 390 nm UV light and a 488 nm blue light, respectively, a nonvolatile holographic recording can be realized. Doping the appropriate Cr (for example, N-Cr = 2.795 X 10(25)m(-3) and N-Cr/N-Cu = 1) benefits the improvement of holographic recording properties. (c) 2005 Optical Society of America.

Improvement of sensitivity and refractive-index changes in LiNbO3 : Ce : Cu crystals with UV light

A nonvolatile recording scheme is proposed using LiNbO3:Ce:Cu crystals and modulated UV light to record gratings simultaneously in two centres and using red light to bleach the grating in the shallow centre to realize persistent photorefractive holographic storage. Compared with the normal UV-sensitized nonvolatile holographic system, the amplitude of refractive-index changes is greatly increased and the recording sensitivity is significantly enhanced by recording with UV light in the LiNbO3:Ce:Cu crystals. Based on jointly solving the two-centre material equations and the coupled-wave equations, temporal evolutions of the photorefractive grating and the diffraction effciency are effectively described and numerically analysed. Roles of doping levels and recording-beam intensity are discussed in detail. Theoretical results confirm and predict experimental results.

Refractive-index change and sensitivity improvement in holographic recording in LiNbO3:Ce:Cu crystals with green light

Nonvolatile holographic recording is performed with green light in LiNbO3:Ce:Cu crystals. The refractive-index change and the recording sensitivity are times better than those obtained by recording with red light, and higher optical fixing efficiency is obtained. Correspondingly, theoretical investigations are given.

Diseño de un detector para una red de vigilancia radiológica ambiental

[ES]El objetivo de este trabajo es analizar el montaje de un contador proporcional de radiación para su uso en una red de vigilancia radiológica ambiental desde su diseño básico hasta su puesta en marcha. Se realizará un análisis de las razones que nos han llevado a elegir este tipo de contadores de ionización gaseosa (frente a otros tipos), para llegar a la conclusión de que son los más adecuados para la vigilancia radiológica ambiental. Este análisis describirá la solución a la que se ha llegado para construir un contador con los medios al alcance y el porqué de su diseño concreto, así como la metodología de trabajo que se ha seguido para la obtención del mismo. Basándonos en suposiciones teóricas derivadas del análisis geométrico de la instrumentación para la obtención de resultados previsibles, se realizará una verificación mediante ensayos técnicos en el laboratorio. Se presupuestará el material utilizado y las horas requeridas, para llevar una cuenta de lo que le ha costado al departamento y lo que le habría costado en caso de no poseer la instrumentación utilizada. Después de realizar diversos test relacionados con la instrumentación electrónica, se procederá a obtener unas conclusiones derivadas de los ensayos en el laboratorio para su posterior comparación con las estimaciones teóricas. En el anexo I se encuentran los planos de diseño que se han realizado y en el anexo II los resultados finales obtenidos de los ensayos.

Diseño y desarrollo de una interfaz de configuración de un generador de tráfico de red

[ES]Se trata de un software de gobierno para un inyector de tráfico de altas prestaciones capaz de adaptar el tráfico de red a distintos parámetros. Además, se deberán integrar el conjunto en la arquitectura de pruebas del laboratorio.

Diseño electrónico de una red de comunicaciones para monitorización de estructuras

[ES]El Trabajo Fin de Grado “Diseño electrónico de una red de comunicaciones para monitorización de estructuras” consiste en el desarrollo de un sistema de comunicación orientado al despliegue y gestión de una red con un alto número de sensores para monitorización de estructuras a bordo de una aeronave. El objetivo principal es hacer viable la utilización de dicha red, partiendo de la premisa de crear una alternativa con mayor eficiencia energética que los sistemas actualmente disponibles para tal fin.

Reposición de la Red Básica Municipal de Donostia/San Sebastián

En el presente Trabajo Fin de Grado, se trabajará sobre la Red Básica Municipal de Donostia/San Sebastián, donde el autor ha sido partícipe en la campaña de reposición de la Red. Nos remontaremos a los orígenes de la Red, explicando cómo nació la Red Básica Municipal de Donostia/San Sebastián y la metodología que se usó para los trabajos previos a su implantación. A lo largo del trabajo, se explicará la metodología de observación y cálculo que se ha empleado en la actual reposición de la Red, el cual ha sido desarrollado por la empresa Geograma S.L. para el Ayuntamiento de San Sebastián entre enero y mayo de 2014. Finalmente, se estudiará como metodología alternativa las observaciones RTK en Red (NRTK), con el fin de validar el método para trabajos de observación de Redes Urbanas de Referencias Topográficas.