New type of dual-channel PAM chlorophyll fluorometer for highly sensitive water toxicity biotests

A new type of dual-channel PAM chlorophyll fluorometer has been developed, which is specialised in the detection of extremely small differences in photosynthetic activity in algae or thylakoids suspensions. In conjunction with standardised algae cultures or isolated thylakoids, the new device provides an ultrasensitive biotest system for detection of toxic substances in water samples. In this report, major features of the new device are outlined and examples of its performance are presented using suspensions of Phaeodactylum tricornutum (diatoms) and of freeze-dried thylakoids of Lactuca sativa (salad). Investigated and reference samples are exposed to the same actinic intensity of pulse-modulated measuring light. The quantum yields are assessed by the saturation pulse method. Clock-triggered repetitive measurements of quantum yield typically display a standard deviation of 0.1%, corresponding to the inhibition induced by 0.02 mug diuron l(-1). Hence, for diuron or compounds with similar toxicity, the detection limit is well below the 0.1 mug l(-1) defined as the limit for the presence of a single toxic substance in water by the European Commission drinking water regulation. The amounts of water and biotest material required for analysis are very small, as a single assay involves two 1 ml samples, each containing ca. 0.5 mug chlorophyll. Both with Phaeodactylum and thylakoids the relationship between inhibition and diuron concentration is strictly linear up to 10% inhibition, with very similar slopes. Apparent inhibition depends on the actinic effect of the measuring light, showing optima at 6 and 4 mumol quanta m(-2) s(-1) with Phaeodactylum and thylakoids, respectively.

Dual-channel electrospray microchip.

A dual-channel electrospray microchip has been developed for electrospray ionization mass spectrometry (ESI-MS) where aqueous samples are mixed at the Taylor cone with an organic buffer. Due to the fast and effective mixing in the Taylor cone, the aqueous sample can be well ionized with a high ion intensity. The influence of geometric parameters such as the distance between the two microchannels at their junction at the tip of the emitter has been investigated together with chemical parameters such as the organic buffer composition.

Simultaneous dual channel phase regeneration in SOAs

For the first time we demonstrate simultaneous suppression of phase distortion on two independent 10.7 Gbit/s DPSK modulated signal wavelengths using semiconductor optical amplifiers, realizing a compact phase sensitive amplifier with low power consumption.

Simultaneous dual channel phase regeneration in SOAs

For the first time we demonstrate simultaneous suppression of phase distortion on two independent 10.7 Gbit/s DPSK modulated signal wavelengths using semiconductor optical amplifiers, realizing a compact phase sensitive amplifier with low power consumption.

Verificación de los niveles de atenuación de dos tipos de protectores auditivos tipo copa en trabajadores de una empresa del sector Hidrocarburos. Barrancabermeja-Colombia 2013.

El presente estudio se realizó con el fin de verificar los niveles de atenuación de dos tipos de protectores auditivos tipo copa, bajo condiciones reales de operación en una empresa del sector hidrocarburos en Colombia. Esta verificación se llevó a cabo determinando las diferencias existentes entre los niveles de atenuación establecidos por el fabricante (NRR Teórico), los niveles de atenuación ajustados de acuerdo a la metodología propuesta por NIOSH (NRR Ajustado) y los niveles de atenuación obtenidos experimentalmente (NRR Experimental). El estudio se desarrolló sobre la evaluación de los niveles equivalentes de presión sonora, mediante dosimetrías de doble canal, valorando los niveles de ruido al interior y al exterior del pabellón auditivo de forma simultánea. Los valores obtenidos en las mediciones se ajustaron a los criterios establecidos en la norma ISO 11904 parte 1 considerando la estimación de niveles de inmisión sonora mediante técnica MIRE (Microphone In a Real Ear) corregidos a condiciones de campo libre. Los resultados obtenidos permitieron establecer diferencias significativas entre los niveles de atenuación teóricos, ajustados y experimentales, aspecto que brinda oportunidades de mejora cuando se trata de seleccionar y mantener elementos de protección personal auditiva como estrategia preventiva primaria en el propósito de la disminución de la probabilidad de ocurrencia de hipoacusia neurosensorial inducida por ruido en el lugar de trabajo.

Microfabricated dual sprayer for on-line mass tagging of phosphopeptides.

Phosphopeptides tagging reactions by dinuclear zinc(II) complexes (1,3-bis[bis(2-pyridylmethyl)amino]-propan-2-olato dizinc(II)3+, called tag) were performed with a dual-channel microsprayer in electrospray ionization mass spectrometry. The reaction is first studied ex situ and analyzed with a commercial electrospray source. In situ reactions (i.e., inside the Taylor cone) were achieved with a dual-channel microsprayer both with the tag synthesized chemically before the experiments and with the tag electrogenerated by in situ oxidation of a zinc electrode, also used to apply the electrospray current. The device consists of a polyimide microchip with two microchannels (20 microm x 50 microm x 1 cm) etched on each side of the structure and connecting only at the tip of the microchip. We demonstrate here that mixing two solutions with different physicochemical properties inside the Taylor cone can be used to selectively tag target molecules.

Small molecule dual-inhibitors of TRPV4 and TRPA1 for attenuation of inflammation and pain.

TRPV4 ion channels represent osmo-mechano-TRP channels with pleiotropic function and wide-spread expression. One of the critical functions of TRPV4 in this spectrum is its involvement in pain and inflammation. However, few small-molecule inhibitors of TRPV4 are available. Here we developed TRPV4-inhibitory molecules based on modifications of a known TRPV4-selective tool-compound, GSK205. We not only increased TRPV4-inhibitory potency, but surprisingly also generated two compounds that potently co-inhibit TRPA1, known to function as chemical sensor of noxious and irritant signaling. We demonstrate TRPV4 inhibition by these compounds in primary cells with known TRPV4 expression - articular chondrocytes and astrocytes. Importantly, our novel compounds attenuate pain behavior in a trigeminal irritant pain model that is known to rely on TRPV4 and TRPA1. Furthermore, our novel dual-channel blocker inhibited inflammation and pain-associated behavior in a model of acute pancreatitis - known to also rely on TRPV4 and TRPA1. Our results illustrate proof of a novel concept inherent in our prototype compounds of a drug that targets two functionally-related TRP channels, and thus can be used to combat isoforms of pain and inflammation in-vivo that involve more than one TRP channel. This approach could provide a novel paradigm for treating other relevant health conditions.

Chlormequat selective electrodes: construction, evaluation and application at FIA systems

A flow injection analysis (FIA) system having a chlormequat selective electrode is proposed. Several electrodes with poly(vinyl chloride) based membranes were constructed for this purpose. Comparative characterization suggestedthe use of membrane with chlormequat tetraphenylborate and dibutylphthalate. On a single-line FIA set-up, operating with 1x10-2 mol L-1 ionic strength and 6.3 pH, calibration curves presented slopes of 53.6±0.4mV decade-1 within 5.0x10-6 and1.0x10-3 mol L-1, andsquaredcorrelation coefficients >0.9953. The detection limit was 2.2x10-6 mol L-1 and the repeatability equal to ±0.68mV (0.7%). A dual-channel FIA manifold was therefore constructed, enabling automatic attainment of previous ionic strength andpH conditions and thus eliminating sample preparation steps. Slopes of 45.5±0.2mV decade -1 along a concentration range of 8.0x10-6 to 1.0x10-3 mol L-1 with a repeatability ±0.4mV (0.69%) were obtained. Analyses of real samples were performed, and recovery gave results ranging from 96.6 to 101.1%.

Use of limitations of radiolabeled anti-CEA antibodies and their fragments for photoscanning detection of human colorectal carcinomas.

Fifty-three patients with histologically proven carcinoma were injected with highly purified [131I]-labeled goat antibodies or fragments of antibodies against carcinoembryonic antigen (CEA). Each patient was tested by external photoscanning 4, 24, 36 and 48 h after injection. In 22 patients (16 of 38 injected with intact antibodies, 5 of 13 with F(ab')2 fragments and 1 of 2 with Fab' fragments), an increased concentration of 131I radioactivity corresponding to the previously known tumor location was detected by photoscanning 36-48 h after injection. Blood pool and secreted radioactivity was determined in all patients by injecting 15 min before scanning, [99mTc]-labeled normal serum albumin and free 99mTc04-. The computerized subtraction of 99mTc from 131I radioactivity enhanced the definition of tumor localization in the 22 positive patients. However, in spite of the computerized subtraction, interpretation of the scans remained doubtful for 12 patients and was entirely negative for 19 additional patients. In order to provide a more objective evaluation for the specificity of the tumor localization of antibodies, 14 patients scheduled for tumor resection were injected simultaneously with [131I]-labeled antibodies or fragments and with [125I]-labeled normal goat IgG or fragments. After surgery, the radioactivity of the two isotopes present either in tumor or adjacent normal tissues was measured in a dual channel scintillation counter. The results showed that the antibodies or their fragments were 2-4 times more concentrated in the tumor than in the normal tissues. In addition, it was shown that the injected antibodies formed immune complexes with circulating CEA and that the amount of immune complexes detectable in serum was roughly proportional to the level of circulating CEA.

Radio-holografía de microondas para la optimización de la superficie de grandes antenas reflectoras

En el presente trabajo de tesis se afronta el problema de la optimización de la superficie de grandes antenas reflectoras. Es sabido que los grandes reflectores, formados por una superficie panelada, sufren deformaciones debidas al impacto del viento, a los cambios de temperatura y a los efectos gravitacionales derivados del gran peso de la estructura. Estos efectos hacen que los reflectores pierdan su forma ideal, generalmente de paraboloide, y se reduzca su eficiencia de apertura y, por tanto, se limite la máxima frecuencia de uso de los mismos. Es necesario, por tanto, disponer de técnicas que permitan medir el estado de la superficie de grandes reflectores, y derivar los ajustes necesarios a aplicar sobre los tornillos de soporte de cada uno de los paneles que conforman dicha superficie. De esta manera, se devolvería al reflector su forma óptima y aumentaría la eficiencia de apertura y el rango de frecuencias de uso. Hay que resaltar que el aumento de la eficiencia de un radiotelescopio supone una reducción en el tiempo de integración necesario para la detección de las debilísimas señales generadas por las radiofuentes naturales, ahorrando así valioso tiempo de observación. Además, el incremento en el rango de frecuencias permite la detección de nuevas líneas o especies moleculares en dichas radiofuentes. Tras un primer capítulo introductorio, se presenta, en el capítulo segundo, la geometría de estos grandes reflectores y la influencia de los distintos factores que afectan a la calidad de la superficie de los mismos, como la gravedad, el viento y la temperatura, particularizando para el caso del radiotelescopio de 40 metros del Centro Astronómico de Yebes. En el tercer capítulo, se presentan las diferentes técnicas metrológicas empleadas actualmente para abordar la determinación de estos ajustes, mostrándose las ventajas e inconvenientes de cada una de ellas. Actualmente, la técnica metrológica más precisa y rápida para llevar a cabo esta tarea de caracterización de la superficie de un gran reflector, es la radio-holografía de microondas presentada en el capítulo cuarto. A partir de las medidas proporcionadas por esta técnica, realizadas con la ayuda de un transmisor, y mediante transformaciones de campo, se calculan los errores de la superficie del reflector, respecto al paraboloide ideal, y se derivan los ajustes necesarios. En los capítulos quinto y sexto se presentan los resultados de la aplicación de esta técnica a dos radiotelescopios: el de 30 metros de IRAM en Pico de Veleta (Granada) y los prototipos de 12 metros de las antenas del proyecto ALMA. Por su parte, el capítulo séptimo contiene el núcleo fundamental de esta tesis y presenta el desarrollo de la técnica de radio-holografía de microondas para optimizar la superficie del radiotelescopio de 40 metros del Centro Astronómico de Yebes. Para ello, ha sido necesario diseñar, construir e instalar un receptor de doble canal en banda Ku en foco primario, y la instrumentación asociada para hacer las medidas de amplitud y fase del diagrama de radiación. Además, ha sido necesario desarrollar el software para llevar a cabo las transformaciones de campo y derivar los ajustes de los paneles. De las medidas holográficas iniciales resultó un error de la superficie del radiotelescopio de 485 μm WRMS, respecto al paraboloide ideal en dirección normal. Tras varias iteraciones del proceso de medida y ajuste, se consiguió reducir dicho error a 194 μm WRMS. Esta notable mejora de la calidad de la superficie ha supuesto aumentar la eficiencia de apertura desde 2,6% al 38,2% a 86 GHz, para un receptor a esta frecuencia situado en el foco primario que produjese la misma iluminación que el receptor de holografía. In this thesis the problem of large reflector antenna surface optimization is faced. It is well known that large reflectors, which are made of a panelled surface, suffer from deformations due to the impact of wind, temperature gradients and gravity loads coming from the high weigth of the structure. These effects distort the ideal reflector shape, which is a paraboloid in most cases, hence reducing the aperture efficiency of the reflector and limiting the maximum frequency of operation. Therefore, it is necessary to have some techniques to measure the status of large reflector surfaces and to derive the adjustment values to be applied to the screws that connect the surface panels to the reflector back-up structure. In this way, the reflector would recover its optimum shape and the aperture efficiency and frequency range would increase. It has to be stated that an increment in the radiotelescope aperture efficiency would imply a reduction in the integration time needed to detect such weak signals coming from natural radiosources in space and, hence, an important saving in observation time. In addition, the increase in the frequency range of operation would allow the detection of new molecular lines in those radiosources. After the introduction, the second chapter shows the geometry of large reflector antennas and the impact on its surface quality of different factors like gravity, wind and temperature, particularly for the case of the Centro Astronómico de Yebes 40 meter radiotelescope. The third chapter deals with the different metrology techniques used to determine the panel adjustments, including the advantages and drawbacks of each one Currently, the most accurate and fast metrologic technique to carry out the characterization of large reflector surfaces is microwave radio-holography2, which is shown in chapter four. From the measurements provided by microwave radio-holography, performed with the help of a transmitter, and with the use of field transformations, the reflector surface errors are computed and the panel adjustments are derived. Chapters five and six show the results of holographic measurements applied to two first class radiotelescopes: the IRAM 30 meter radiotelescope and the 12 meter prototype antennas for the ALMA project. Chapter seven contains the main work of this thesis. It presents the development of the microwave radio-holography technique for the optimization of the Centro Astronómico de Yebes 40m radiotelescope. The work implied the design, construction and instalation of a prime focus Ku-band dual channel receiver, together with the associated instrumentation to measure the amplitude and phase of the radiotelescope radiation pattern. In addition, the software to carry out field transformations and screw settings computations was developed too. Initial holography measurements came up with an surface error of 485 μmWRMS in normal direction with respect to the best-fit paraboloid. After a few iterations of the measurementadjustment cycle, the surface error was reduced to 194 μm WRMS. This remarkable improvement in surface quality means an increment in aperture efficiency from 2,6% to 38,2% at 86 GHz, assuming a receiver at this frequency in prime focus position which produces the same illumination as the holography receiver.

The selection of a model microalgal species as biomaterial for a novel aquatic phytotoxicity assay

A phytotoxicity assay based on the ToxY-PAM dual-channel yield analyser has been developed and successfully incorporated into field assessments for the detection of phytotoxicants in water. As a means of further exploring the scope of the assay application and of selecting a model biomaterial to complement the instrument design, nine algal species were exposed to four chemical substances deemed of priority for water quality monitoring purposes (chlorpyrifos, copper, diuron and nonylphenol ethoxylate). Inter-species differences in sensitivity to the four toxicants varied by a factor of 1.9-100. Measurements of photosystem-II quantum yield using these nine single-celled microalgae as biomaterial corroborated previous studies which have shown that the ToxY-PAM dual-channel yield analyser is a highly sensitive method for the detection of PS-II impacting herbicides. Besides Phaeodactylum tricornutum, the previously applied biomaterial, three other species consistently performed well (Nitzschia closterium, Chlorella vulgaris and Dunaliella tertiolecta) and will be used in further test optimisation experiments. In addition to sensitivity, response time was evaluated and revealed a high degree of variation between species and toxicants. While most species displayed relatively weak and slow responses to copper, C. vulgaris demonstrated an IC10 of 51 μ g L-1, with maximum response measured within 25 minutes and inhibition being accompanied by a large decrease in fluorescence yield. The potential for this C vulgaris-based bioassay to be used for the detection of copper is discussed. There was no evidence that the standard ToxY-PAM protocol, using these unicellular algae species, could be used for the detection of chlorpyrifos or nonylphenol ethoxylate at environmentally relevant levels. © 2005 Elsevier B.V. All rights reserved.

The combined SPE : ToxY-PAM phytotoxicity assay; application and appraisal of a novel biomonitoring tool for the aquatic environment

Mounting concerns regarding the environmental impact of herbicides has meant a growing requirement for accurate, timely information regarding herbicide residue contamination of, in particular, aquatic systems. Conventional methods of detection remain limited in terms of practicality due to high costs of operation and the specialised information that analysis provides. A new phytotoxicity bioassay was trialled for the detection of herbicide residues in filter-purified (Milli-Q) as well as natural waters. The performance of the system, which combines solid-phase extraction (SPE) with the ToxY-PAM dual-channel yield analyser (Heinz Walz GmbH), was tested alongside the traditional method of liquid chromatography-mass spectrometry (LC-MS). The assay methodology was found to be highly sensitive (LOD 0.1 ng L-1 diuron) with good reproducibility. The study showed that the assay protocol is time effective and can be employed for the aquatic screening of herbicide residues in purified as well as natural waters.

Comparative performance of analytical techniques for polyhexamethylene biguanide (PHMB) in ophthalmic solutions

Purpose: The use of PHMB as a disinfectant in contact lens multipurpose solutions has been at the centre of much debate in recent times, particularly in relation to the issue of solution induced corneal staining. Clinical studies have been carried out which suggest different effects with individual contact lens materials used in combination with specific PHMB containing care regimes. There does not appear to be, however, a reliable analytical technique that would detect and quantify with any degree of accuracy the specific levels of PHMB that are taken up and released from individual solutions by the various contact lens materials. Methods: PHMB is a mixture of positively charged polymer units of varying molecular weight that has maximum absorbance wavelength of 236 nm. On the basis of these properties a range of assays including capillary electrophoresis, HPLC, a nickelnioxime colorimetric technique, mass spectrophotometry, UV spectroscopy and ion chromatography were assessed paying particular attention to each of their constraints and detection levels. Particular interest was focused on the relative advantage of contactless conductivity compared to UV and mass spectrometry detection in capillary electrophoresis (CE). This study provides an overview of the comparative performance of these techniques. Results: The UV absorbance of PHMB solutions, ranging from 0.0625 to 50 ppm was measured at 236 nm. Within this range the calibration curve appears to be linear however, absorption values below 1 ppm (0.0001%) were extremely difficult to reproduce. The concentration of PHMB in solutions is in the range of 0.0002–0.00005% and our investigations suggest that levels of PHMB below 0.0001% (levels encountered in uptake and release studies) can not be accurately estimated, in particular when analysing complex lens care solutions which can contain competitively absorbing, and thus interfering, species in the solution. The use of separative methodologies, such as CE using UV detection alone is similarly limited. Alternative techniques including contactless conductivity detection offer greater discrimination in complex solutions together with the opportunity for dual channel detection. Preliminary results achieved by TraceDec1 contactless conductivity detection, (Gain 150%, Offset 150) in conjunction with the Agilent capillary electrophoresis system using a bare fused silica capillary (extended light path, 50 mid, total length 64.5 cm, effective length 56 cm) and a cationic buffer at pH 3.2, exhibit great potential with reproducible PHMB split peaks. Conclusions: PHMB-based solutions are commonly associated with the potential to invoke corneal staining in combination with certain contact lens materials. However this terminology ‘PHMBbased solution’ is used primarily because PHMB itself has yet to be adequately implicated as the causative agent of the staining and compromised corneal cell integrity. The lack of well characterised adequately sensitive assays, coupled with the range of additional components that characterise individual care solutions pose a major barrier to the investigation of PHMB interactions in the lenswearing eye.

Blind adaptive channel estimation for dual-rate DS/CDMA signals

This letter proposes the subspace-based blind adaptive channel estimation algorithm for dual-rate quasi-synchronous DS/CDMA systems, which can operate at the low-rate (LR) or high-rate (HR) mode. Simulation results show that the proposed blind adaptive algorithm at the LR mode has a better performance than that at the HR mode, with the cost of an increasing computational complexity.

Blind channel estimation for dual-rate DS/CDMA

This paper proposes a subspace based blind adaptive channel estimation algorithm for dual-rate DS-CDMA systems, which can operate at the low-rate (LR) or high-rate (HR) mode. Simulation results show that the proposed blind adaptive algorithm at the LR mode has a better performance than that at the HR mode, with the cost of an increased computational complexity.