Testing of fluorescent DC lamps for Solar Home Systems

With the advent of the Universal Technical Standard for Solar Home Systems, procedures to test the compliance of SHS fluorescent lamps with the standard have been developed. Definition of the laboratory testing procedures is a necessary step in any lamp quality assurance procedure. Particular attention has been paid to test simplicity and to affordability, in order to facilitate local application of the testing procedures, for example by the organisations which carry out electrification programmes. The set of test procedures has been applied to a representative collection of 42 lamps from many different countries, directly acquired in the current photovoltaic rural electrification market. Tests apply to: lamp resistance under normal operating conditions; lamp reliability under extreme conditions; under abnormal conditions; and lamp luminosity. Results are discussed and some recommendations for updating the relevant standard are given. The selected technical standard, together with the proposed testing procedures, form the basis of a complete quality assurance tool that can be applied locally in normal electrical laboratories. Full testing of a lamp requires less than one month, which is very reasonable on the context of quality assurance programmes

The effect of acoustic screens on orchestra musicians

La Directiva 2003/10/CE del Parlamento Europeo y del Consejo, del 6 de febrero de 2003, específica con arreglo al apartado 1 del artículo 16 de la Directiva 89/391/CEE las disposiciones mínimas de seguridad y de salud relativas a la exposición de los trabajadores a los riesgos derivados de los agentes físicos (ruido). En la industria musical, y en concreto en los músicos de orquesta, una exposición de más de ocho horas al día a un nivel de presión sonora de 80dB(A) o más es algo muy común. Esta situación puede causar a los trabajadores daños auditivos como la hiperacusia, hipoacusia, tinitus o ruptura de la membrana basilar entre otros. Esto significa que deben tomarse medidas para implementar las regulaciones de la forma más razonable posible para que la interpretación del músico, la dinámica y el concepto musical que se quiere transmitir al público se vea lo menos afectada posible. Para reducir la carga auditiva de los músicos de orquesta frente a fuertes impactos sonoros provenientes de los instrumentos vecinos, se está investigando sobre el uso de unos paneles acústicos que colocados en puntos estratégicos de la orquesta pueden llegar a reducir el impacto sonoro sobre el oído hasta 20dB. Los instrumentos de viento metal y de percusión son los responsables de la mayor emisión de presión sonora. Para proteger el oído de los músicos frente a estos impactos, se colocan los paneles en forma de barrera entre dichos instrumentos y los músicos colocados frente a ellos. De esta forma se protege el oído de los músicos más afectados. Para ver el efecto práctico que producen estos paneles en un conjunto orquestal, se realizan varias grabaciones en los ensayos y conciertos de varias orquestas. Los micrófonos se sitúan a la altura del oído y a una distancia de no más de 10cm de la oreja de varios de los músicos más afectados y de los músicos responsables de la fuerte emisión sonora. De este modo se puede hacer una comparación de los niveles de presión sonora que percibe cada músico y evaluar las diferencias de nivel existentes entre ambos. Así mismo se utilizan configuraciones variables de los paneles para comparar las diferencias de presión sonora que existen entre las distintas posibilidades de colocarlos y decidir así sobre la mejor ubicación y configuración de los mismos. A continuación, una vez obtenidos las muestras de audio y los diferentes archivos de datos medidos con un analizador de audio en distintas posiciones de la orquesta, todo ello se calibra y analiza utilizando un programa desarrollado en Matlab, para evaluar el efecto de los paneles sobre la percepción auditiva de los músicos, haciendo especial hincapié en el análisis de las diferencias de nivel de presión sonora (SPL). Mediante el cálculo de la envolvente de las diferencias de nivel, se evalúa de un modo estadístico el efecto de atenuación de los paneles acústicos en los músicos de orquesta. El método está basado en la probabilidad estadística de varias muestras musicales ya que al tratarse de música tocada en directo, la dinámica y la sincronización entre los músicos varía según el momento en que se toque. Estos factores junto con el hecho de que la partitura de cada músico es diferente dificulta la comparación entre dos señales grabadas en diferentes puntos de la orquesta. Se necesita por lo tanto de varias muestras musicales para evaluar el efecto de atenuación de los paneles en las distintas configuraciones mencionadas anteriormente. El estudio completo del efecto de los paneles como entorno que influye en los músicos de orquesta cuando están sobre el escenario, tiene como objetivo la mejora de sus condiciones de trabajo. Abstract For several years, the European Union has been adopting many laws and regulations to protect and give more security to people who are exposed to some risk in their job. Being exposed to a loud sound pressure level during many hours in the job runs the risk of hearing damage. Particularly in the field of music, the ear is the most important working tool. Not taking care of the ear can cause some damage such as hearing loss, tinnitus, hyperacusis, diplacusis, etc. This could have an impact on the efficiency and satisfaction of the musicians when they are playing, which could also cause stress problems. Orchestra musicians, as many other workers in this sector, are usually exposed to a sound level of 80dB(A) or more during more than eight hours per day. It means that they must satisfy the law and their legal obligations to avoid health problems proceeding from their job. Putting into practice the new regulations is a challenge for orchestras. They must make sure that the repertoire, with its dynamic, balance and feeling, is not affected by the reduction of sound levels imposed by the law. This study tries to investigate the benefits and disadvantages of using shields as a hearing protector during rehearsals and orchestral concerts.

Emissions and economic costs of cycling compact fluorescent lamps with integrated ballasts

This paper proposes a way to quantify the emissions of mercury (Hg) and CO2 associated with the manufacture and operation of compact fluorescent lamps with integrated ballasts (CFLis), as well as the economic cost of using them under different operating cycles. The main purpose of this paper is to find simple criteria for reducing the polluting emissions under consideration and the economic cost of CFLi to a minimum. A lifetime model is proposed that allows the emissions and costs to be described as a function of degradation from turning CFLi on and their continuous operation. An idealized model of a CFLi is defined that combines characteristics stated by different manufacturers. In addition, two CFLi models representing poor-quality products are analyzed. It was found that the emissions and costs per unit of time of operation of the CFLi depend linearly on the number of times per unit of time it is turned on and the time of continuous operation. The optimal conditions (lowest emissions and costs) depend on the place of manufacture, the place of operation and the quality of the components of the lamp/ballast. Finally, it was also found that for each lamp, there are intervals when it is turned off during which emissions of pollutants and costs are identical regardless of how often the lamp is turned on or the time it remains on. For CO2 emissions, the lamp must be off up to 5 minutes; for the cost, up to 7 minutes and for Hg emissions, up to 43 minutes. It is advisable not to turn on a CFLi sooner than 43 minutes from the last time it was turned off.

Acoustic characterisation of cinema screens

As a consequence of cinema screens being placed in front of screen-speakers, a reduction in sound quality has been noticed. Cinema screens not only let the sound go through them, but also absorb a small amount of it and reflect the sound which impacts on the screen to the back, coming forward again in case it impacts on the loudspeaker. This backwards reflection in addition to the signal coming from the loudspeaker can lead to constructive or destructive interference at certain frequencies which usually results in comb filtering. In this project, this effect has been studied through researching amongst various data sheet provided by different manufacturers, acoustical measurements completed in the large anechoic chamber of the ISVR and some theoretical models developed with MatLab software. If results obtained with MatLab are accurate enough in comparison to the real measurements taken in the anechoic chamber this would lead to a good way to predict which would be the attenuation added to the system at each frequency, given that not all manufacturers provide an attenuation curve, but only an average attenuation. This average attenuation might be useless as sound waves have different wavelengths and its propagation through partitions varies. In fact, sound is composed by high and low frequencies, where high frequencies are characterised by a small wavelength which is usually easier to attenuate than low frequencies that characterised by bigger wavelengths. Furthermore, this information would be of great value to both screen manufacturers, who could offer a much more precise data in their data sheets; and customers, who would have a great amount of information to their disposal before purchasing and installing anything in their cinemas, being able to know by themselves which screen or loudspeaker should be best to meet their expectative. RESUMEN. La aparición de la digitalización de las bandas sonoras para las películas hace posible la mejora en la calidad de sonido de los cines. Sin embargo, un aspecto a tener en cuenta en esta calidad del sonido es la transmisión de éste a través de la pantalla, ya que normalmente tras ella se encuentran situados los altavoces. Las propiedades acústicas varían dependiendo del tipo de pantalla que se utilice, además de haber poca información a la que acceder para poder valorar su comportamiento. A lo largo de este proyecto, se analizan tres muestras de pantallas distintas donadas por distintos fabricantes para poder llegar a la conclusión de dependiendo del tipo de pantalla cuál es la distancia óptima a la que localizar la pantalla respecto al altavoz y con qué inclinación. Dicho análisis se realizó en la cámara anecoica del ISVR (University of Southampton) mediante la construcción de un marco de madera de 2x2 m en el que tensar las pantallas de cine, y un altavoz cuyo comportamiento sea el más similar al de los altavoces de pantalla reales. Los datos se captaron mediante cuatro micrófonos colocados en posiciones distintas y conectados al software Pulse de Brüel & Kjær, a través del cual se obtuvieron las respuestas en frecuencia del altavoz sin pantalla y con ella a diferentes distancias del altavoz. Posteriormente, los datos se analizaron con MatLab donde se calculó la atenuación, el factor de transmisión de la presión (PTF) y el análisis cepstrum. Finalmente, se realizó un modelo teórico del comportamiento de las pantallas perforadas basado en las placas perforadas utilizadas para atenuar el sonido entre distintas habitaciones. Como conclusión se llegó a que las pantallas curvadas son acústicamente más transparentes que las pantallas perforadas que a partir de 6 kHz son más acústicamente opacas. En las pantallas perforadas la atenuación depende del número de perforaciones por unidad de área y el diámetro de éstas. Dicha atenuación se reducirá si se reduce el diámetro de las perforaciones de la pantalla, o si se incrementa la cantidad de perforaciones. Acerca del efecto filtro peine, para obtener la mínima amplitud de éste la pantalla se deberá situar a una distancia entre 15 y 30 cm del altavoz, encontrando a la distancia de 30 cm que la última reflexión analizada a través de Cepstrum llega 5 ms más tarde que la señal directa, por lo cual no debería dañar el sonido ni la claridad del habla.

Reconfiguración de un espacio con pantallas de teja = Reconfiguring a space using roofing tile screens

Artículo a partir de extracto de la memoria de proyecto de la rehabilitación de la nave 8B de Matadero Madrid

Measurement of cytosolic, mitochondrial, and Golgi pH in single living cells with green fluorescent proteins

Many cellular events depend on a tightly compartmentalized distribution of H+ ions across membrane-bound organelles. However, measurements of organelle pH in living cells have been scarce. Several mutants of the Aequorea victoria green fluorescent protein (GFP) displayed a pH-dependent absorbance and fluorescent emission, with apparent pKa values ranging from 6.15 (mutations F64L/S65T/H231L) and 6.4 (K26R/F64L/S65T/Y66W/N146I/M153T/V163A/N164H/H231L) to a remarkable 7.1 (S65G/S72A/T203Y/H231L). We have targeted these GFPs to the cytosol plus nucleus, the medial/trans-Golgi by fusion with galactosyltransferase, and the mitochondrial matrix by using the targeting signal from subunit IV of cytochrome c oxidase. Cells in culture transfected with these cDNAs displayed the expected subcellular localization by light and electron microscopy and reported local pH that was calibrated in situ with ionophores. We monitored cytosolic and nuclear pH of HeLa cells, and mitochondrial matrix pH in HeLa cells and in rat neonatal cardiomyocytes. The pH of the medial/trans-Golgi was measured at steady-state (calibrated to be 6.58 in HeLa cells) and after various manipulations. These demonstrated that the Golgi membrane in intact cells is relatively permeable to H+, and that Cl− serves as a counter-ion for H+ transport and likely helps to maintain electroneutrality. The amenability to engineer GFPs to specific subcellular locations or tissue targets using gene fusion and transfer techniques should allow us to examine pH at sites previously inaccessible.

Mitochondrial localization of a NADR-dependent isocitrate dehydrogenase isoenzyme by using the green fluorescent protein as a marker

In this work, we describe the isolation of a new cDNA encoding an NADP-dependent isocitrate dehydrogenase (ICDH). The nucleotide sequence in its 5′ region gives a deduced amino acid sequence indicative of a targeting peptide. However, even if this cDNA clearly encodes a noncytosolic ICDH, it is not possible to say from the targeting peptide sequence to which subcellular compartment the protein is addressed. To respond to this question, we have transformed tobacco plants with a construct containing the entire targeting signal-encoding sequence in front of a modified green fluorescent protein (GFP) gene. This construct was placed under the control of the cauliflower mosaic virus 35S promoter, and transgenic tobacco plants were regenerated. At the same time, and as a control, we also have transformed tobacco plants with the same construct but lacking the nucleotide sequence corresponding to the ICDH-targeting peptide, in which the GFP is retained in the cytoplasm. By optical and confocal microscopy of leaf epiderm and Western blot analyses, we show that the putative-targeting sequence encoded by the cDNA addresses the GFP exclusively into the mitochondria of plant cells. Therefore, we conclude that this cDNA encodes a mitochondrial ICDH.

PRIM: Proximity imaging of green fluorescent protein-tagged polypeptides

We report a serendipitous discovery that extends the impressive catalog of reporter functions performed by green fluorescent protein (GFP) or its derivatives. When two GFP molecules are brought into proximity, changes in the relative intensities of green fluorescence emitted upon excitation at 395 vs. 475 nm result. These spectral changes provide a sensitive ratiometric index of the extent of self-association that can be exploited to quantitatively image homo-oligomerization or clustering processes of GFP-tagged proteins in vivo. The method, which we term proximity imaging (PRIM), complements fluorescence resonance energy transfer between a blue fluorescent protein donor and a GFP acceptor, a powerful method for imaging proximity relationships between different proteins. However, unlike fluorescence resonance energy transfer (which is a spectral interaction), PRIM depends on direct contact between two GFP modules, which can lead to structural perturbations and concomitant spectral changes within a module. Moreover, the precise spatial arrangement of the GFP molecules within a given dimer determines the magnitude and direction of the spectral change. We have used PRIM to detect FK1012-induced dimerization of GFP fused to FK506-binding protein and clustering of glycosylphosphatidylinositol-anchored GFP at cell surfaces.

Colocalization of cell division proteins FtsZ and FtsA to cytoskeletal structures in living Escherichia coli cells by using green fluorescent protein

In the current model for bacterial cell division, FtsZ protein forms a ring that marks the division plane, creating a cytoskeletal framework for the subsequent action of other proteins such as FtsA. This putative protein complex ultimately generates the division septum. Herein we report that FtsZ and FtsA proteins tagged with green fluorescent protein (GFP) colocalize to division-site ring-like structures in living bacterial cells in a visible space between the segregated nucleoids. Cells with higher levels of FtsZ–GFP or with FtsA–GFP plus excess wild-type FtsZ were inhibited for cell division and often exhibited bright fluorescent spiral tubules that spanned the length of the filamentous cells. This suggests that FtsZ may switch from a septation-competent localized ring to an unlocalized spiral under some conditions and that FtsA can bind to FtsZ in both conformations. FtsZ–GFP also formed nonproductive but localized aggregates at a higher concentration that could represent FtsZ nucleation sites. The general domain structure of FtsZ–GFP resembles that of tubulin, since the C terminus of FtsZ is not required for polymerization but may regulate polymerization state. The N-terminal portion of Rhizobium FtsZ polymerized in Escherichia coli and appeared to copolymerize with E. coli FtsZ, suggesting a degree of interspecies functional conservation. Analysis of several deletions of FtsA–GFP suggests that multiple segments of FtsA are important for its localization to the FtsZ ring.

Chemical synthesis of the precursor molecule of the Aequorea green fluorescent protein, subsequent folding, and development of fluorescence

The present paper describes the total chemical synthesis of the precursor molecule of the Aequorea green fluorescent protein (GFP). The molecule is made up of 238 amino acid residues in a single polypeptide chain and is nonfluorescent. To carry out the synthesis, a procedure, first described in 1981 for the synthesis of complex peptides, was used. The procedure is based on performing segment condensation reactions in solution while providing maximum protection to the segment. The effectiveness of the procedure has been demonstrated by the synthesis of various biologically active peptides and small proteins, such as human angiogenin, a 123-residue protein analogue of ribonuclease A, human midkine, a 121-residue protein, and pleiotrophin, a 136-residue protein analogue of midkine. The GFP precursor molecule was synthesized from 26 fully protected segments in solution, and the final 238-residue peptide was treated with anhydrous hydrogen fluoride to obtain the precursor molecule of GFP containing two Cys(acetamidomethyl) residues. After removal of the acetamidomethyl groups, the product was dissolved in 0.1 M Tris⋅HCl buffer (pH 8.0) in the presence of DTT. After several hours at room temperature, the solution began to emit a green fluorescence (λmax = 509 nm) under near-UV light. Both fluorescence excitation and fluorescence emission spectra were measured and were found to have the same shape and maxima as those reported for native GFP. The present results demonstrate the utility of the segment condensation procedure in synthesizing large protein molecules such as GFP. The result also provides evidence that the formation of the chromophore in GFP is not dependent on any external cofactor.

Dynamics of fluorescence fluctuations in green fluorescent protein observed by fluorescence correlation spectroscopy

We have investigated the pH dependence of the dynamics of conformational fluctuations of green fluorescent protein mutants EGFP (F64L/S65T) and GFP-S65T in small ensembles of molecules in solution by using fluorescence correlation spectroscopy (FCS). FCS utilizes time-resolved measurements of fluctuations in the molecular fluorescence emission for determination of the intrinsic dynamics and thermodynamics of all processes that affect the fluorescence. Fluorescence excitation of a bulk solution of EGFP decreases to zero at low pH (pKa = 5.8) paralleled by a decrease of the absorption at 488 nm and an increase at 400 nm. Protonation of the hydroxyl group of Tyr-66, which is part of the chromophore, induces these changes. When FCS is used the fluctuations in the protonation state of the chromophore are time resolved. The autocorrelation function of fluorescence emission shows contributions from two chemical relaxation processes as well as diffusional concentration fluctuations. The time constant of the fast, pH-dependent chemical process decreases with pH from 300 μs at pH 7 to 45 μs at pH 5, while the time-average fraction of molecules in a nonfluorescent state increases to 80% in the same range. A second, pH-independent, process with a time constant of 340 μs and an associated fraction of 13% nonfluorescent molecules is observed between pH 8 and 11, possibly representing an internal proton transfer process and associated conformational rearrangements. The FCS data provide direct measures of the dynamics and the equilibrium properties of the protonation processes. Thus FCS is a convenient, intrinsically calibrated method for pH measurements in subfemtoliter volumes with nanomolar concentrations of EGFP.

Trafficking, Assembly, and Function of a Connexin43-Green Fluorescent Protein Chimera in Live Mammalian CellsV⃞

To examine the trafficking, assembly, and turnover of connexin43 (Cx43) in living cells, we used an enhanced red-shifted mutant of green fluorescent protein (GFP) to construct a Cx43-GFP chimera. When cDNA encoding Cx43-GFP was transfected into communication-competent normal rat kidney cells, Cx43-negative Madin–Darby canine kidney (MDCK) cells, or communication-deficient Neuro2A or HeLa cells, the fusion protein of predicted length was expressed, transported, and assembled into gap junctions that exhibited the classical pentalaminar profile. Dye transfer studies showed that Cx43-GFP formed functional gap junction channels when transfected into otherwise communication-deficient HeLa or Neuro2A cells. Live imaging of Cx43-GFP in MDCK cells revealed that many gap junction plaques remained relatively immobile, whereas others coalesced laterally within the plasma membrane. Time-lapse imaging of live MDCK cells also revealed that Cx43-GFP was transported via highly mobile transport intermediates that could be divided into two size classes of <0.5 μm and 0.5–1.5 μm. In some cases, the larger intracellular Cx43-GFP transport intermediates were observed to form from the internalization of gap junctions, whereas the smaller transport intermediates may represent other routes of trafficking to or from the plasma membrane. The localization of Cx43-GFP in two transport compartments suggests that the dynamic formation and turnover of connexins may involve at least two distinct pathways.

The Movement of Coiled Bodies Visualized in Living Plant Cells by the Green Fluorescent Protein

Coiled bodies are nuclear organelles that contain components of at least three RNA-processing pathways: pre-mRNA splicing, histone mRNA 3′- maturation, and pre-rRNA processing. Their function remains unknown. However, it has been speculated that coiled bodies may be sites of splicing factor assembly and/or recycling, play a role in histone mRNA 3′-processing, or act as nuclear transport or sorting structures. To study the dynamics of coiled bodies in living cells, we have stably expressed a U2B"–green fluorescent protein fusion in tobacco BY-2 cells and in Arabidopsis plants. Time-lapse confocal microscopy has shown that coiled bodies are mobile organelles in plant cells. We have observed movements of coiled bodies in the nucleolus, in the nucleoplasm, and from the periphery of the nucleus into the nucleolus, which suggests a transport function for coiled bodies. Furthermore, we have observed coalescence of coiled bodies, which suggests a mechanism for the decrease in coiled body number during the cell cycle. Deletion analysis of the U2B" gene construct has shown that the first RNP-80 motif is sufficient for localization to the coiled body.

Real-Time Imaging of the Axonal Transport of Granules Containing a Tissue Plasminogen Activator/Green Fluorescent Protein Hybrid

A hybrid protein, tPA/GFP, consisting of rat tissue plasminogen activator (tPA) and green fluorescent protein (GFP) was expressed in PC12 cells and used to study the distribution, secretory behavior, and dynamics of secretory granules containing tPA in living cells with a neuronal phenotype. High-resolution images demonstrate that tPA/GFP has a growth cone-biased distribution in differentiated cells and that tPA/GFP is transported in granules of the regulated secretory pathway that colocalize with granules containing secretogranin II. Time-lapse images of secretion reveal that secretagogues induce substantial loss of cellular tPA/GFP fluorescence, most importantly from growth cones. Time-lapse images of the axonal transport of granules containing tPA/GFP reveal a surprising complexity to granule dynamics. Some granules undergo canonical fast axonal transport; others move somewhat more slowly, especially in highly fluorescent neurites. Most strikingly, granules traffic bidirectionally along neurites to an extent that depends on granule accumulation, and individual granules can reverse their direction of motion. The retrograde component of this bidirectional transport may help to maintain cellular homeostasis by transporting excess tPA/GFP back toward the cell body. The results presented here provide a novel view of the axonal transport of secretory granules. In addition, the results suggest that tPA is targeted for regulated secretion from growth cones of differentiated cells, strategically positioning tPA to degrade extracellular barriers or to activate other barrier-degrading proteases during axonal elongation.