Production and characterisation of monoclonal antibodies specific for bovine interleukin-4

Genetic immunisation is a simple method for producing polyclonal antibodies in mice. By this method, we produced antibodies against bovine interleukin-4 (BoIL-4). After a final injection with a recombinant BoIL-4 protein, nine stable hybridoma cell lines were established which secreted monoclonal antibodies (MAbs) against this cytokine. Specific binding of each of the MAbs to recombinant BoIL-4 produced by Escherichia coli, baculovirus, and Trypanosoma brucei was demonstrated in an indirect ELISA and/or in Western blotting. These MAbs recognise the same antigenic region localised in the first 47 amino acids of the mature protein. None of them was able to neutralise the biological activity of the BoIL-4 under the conditions tested but one allowed the detection of BoIL-4 by flow cytometry.

STUDIES ON THE BIOLOGICAL ACTIVITY OF MACROMOLECULES MICROINJECTED INTO MAMMALIAN SOMATIC CELLS

Red Blood cell mediated and glass needle mediated microinjection technology was used to introduce macromolecules into mammalian somatic cells. The biological activities of DNA synthesis inducing factor(s) (Chapter 1), mitotic factor(s) (Chapter 2), and DNA coding for ovalbumin and thymidine kinase (Chapter 3) were studied following injection into mammalian somatic cells.^ Chapter 1. A cell undergoing DNA replication (S phase) contains a factor(s) that induces DNA synthesis prematurely in a G(,1) nucleus when an S phase cell is fused to a G(,1) cell. An assay for the active factor(s) was developed in which a mixture of s phase extract loaded red blood cells (RBC) and synchronous G(,1) HeLa cells was centrifuged onto Concanavalin A (Con A) treated coverslips and fused by PEG. This technique is called "Centrifusion". The synchronous G(,1) HeLa cells injected with S phase extract initiated DNA synthesis earlier than the control G(,1) cells mock injected with RBC loaded with buffer.^ Chapter 2. It has been demonstrated that fusion between a mitotic and an interphase cell usually leads to breakdown of the interphase nucleus, followed by condensation of the interphase chromatin into discrete chromosomes, a process termed premature chromosome condensation. I wanted to develop an assay for the mitotic factor(s) that induces premature chromosome condensation. Experiments were performed utilizing glass needle mediated microinjection of HeLa cell mitotic extract into interphase somatic mammalian cells in an attempt to induce premature chromosome condensation. However, I was not able to induce premature chromosome condensation in the interphase cells, probably because of an inability to introduce sufficient mitotic factor(s) into the cells.^ Chapter 3. A recombinant plasmid containing the chicken ovalbumin gene and three copies of the Herpes thymidine Kinase gene (pOV12-TK) was introduced into mouse LMTK('-) cell nuclei using glass needle mediated gene transfer resulting in LMTK('+) clones that were selected for in HAT medium. Restriction enzyme analysis of the high molecular weight DNA from 6 HAT medium survivor cell clones revealed the presence of one or at best only a few copies of the 12kb ovalbumin gene per mouse genome. Further analysis showed the ovalbumin DNA was not rearranged and was associated with high molecular weight mouse cell DNA. Each of the analyzed cell clones produced ovalbumin demonstrating that the biological activity of the microinjected ovalbumin was retained. ^

Isotope characterisation of Villars Cave stalagmites

We present a new mid-latitude speleothem record of millennial-scale climatic variability during OIS3 from the Villars Cave that, combined with former published contemporaneous samples from the same cave, gives a coherent image of the climate variability in SW-France between ~55 ka and ~30 ka. The 0.82 m long stalagmite Vil-stm27 was dated with 26 TIMS U-Th analyses and its growth curve displays variations that are linked with the stable isotopes, both controlled by the climatic conditions. It consists in a higher resolved replicate of the previously published Vil-stm9 and Vil-stm14 stalagmites where Dansgaard-Oeschger (DO) events have been observed. The good consistency between these three stalagmites and the comparison with other palaoeclimatic reconstructions, especially high resolution pollen records (ODP 976 from the Alboran Sea, Monticchio Lake record from southern Italy) and the nearby MD04-2845 Atlantic Ocean record, permits to draw a specific climatic pattern in SW-France during the OIS3 and to see regional differences between these sites. Main features of this period are: 1) warm events corresponding to Greenland Interstadials (GIS) that are characterized by low speleothem d13C, high temperate pollen percentages, warm temperatures and high humidity; among these events, GIS#12 is the most pronounced one at Villars characterized by an abrupt onset at ~46.6 ka and a duration of about 2.5 ka. The other well individualized warm event coincides with GIS#8 which is however much less pronounced and occurred during a cooler period as shown by a lower growth rate and a higher d13C; 2) cold events corresponding to Greenland Stadials (GS) that are clearly characterized by high speleothem d13C, low temperate pollen abundance, low temperature and enhanced dryness, particularly well expressed during GS coinciding with Heinrich events H5 and H4. The main feature of the Villars record is a general cooling trend between the DO#12 event ~45.5 ka and the synchronous stop of the three stalagmites at ~30 ka ±1, with a first well marked climatic threshold at ~41 ka after which the growth rate and the diameter of all stalagmites slows down significantly. This climatic evolution differs from that shown at southern Mediterranean sites where this trend is not observed. The ~30 ka age marks the second climatic threshold after which low temperatures and low rainfalls prevent speleothem growth in the Villars area until the Lateglacial warming that occurred at ~16.5 ± 0.5 ka. This 15 ka long hiatus, as the older Villars growth hiatus that occurred between 67.4 and 61 ka, are linked to low sea levels, reduced ocean circulation and a southward shift of the Polar Front that likely provoked local permafrost formation. These cold periods coincide with both low summer 65°N insolation, low atmospheric CO2 concentration and large ice sheets development (especially the Fennoscandian).

Mössbauer characterisation of sediments from ODP Sites 128-798 and 128-799

57Fe Mössbauer spectra for 26 sediment and 6 carbonate concretion samples from Sites 798 and 799 were recorded at 293 K. Most spectra were deconvolved to two quadrupole doublets without magnetic hyperfine structure. Typical Mössbauer parameters were: isomer shift (I.S.) = 0.34 mm/s and quadrupole splitting (Q.S.) = 0.64 mm/s for the paramagnetic Fe3+ component (partly, pyrite); I.S. = 1.13 mm/s and Q.S. = 2.64 mm/s for the high-spin Fe2+ component derived from iron-bearing aluminosilicates. A few spectra included other high-spin Fe2+ components ascribed to iron-bearing carbonate minerals (e.g., ferroan magnesite), according to the Mössbauer parameters for Fe2+ in the carbonate concretions. We present the distribution of iron among different chemical forms as a function of depth. These data might indicate changes of depositional and diagenetic conditions.

A characterisation of passive and active interactions and their influence on student's achievement using Moodle LMS Logs

During the last years, there has been much concern about learning management systems' (LMS) effectiveness when compared to traditional learning and about how to assess students' participation during the course. The tracking and monitoring capabilities of most recent LMS have made it possible to analyse every interaction in the system. The issues addressed on this study are: a) Is LMS student's interaction an indicator of academic performance?; b) Are different results in performance expected between distance and in-class LMS-supported education?; c) How can LMS interactions from logs be categorised?; d) May this categorisation detect 'learning witnesses'? To answer these questions, a set of interaction types from Moodle LMS activity record logs has been analysed during two years in online and in-class Master's degrees at the UPM. The results show partial or no evidence of influence between interaction indicators and academic performance, although the proposed categorisation may help detect learning witnesses.

Limiting properties in the characterisation of mixed recycled aggregates for use in the manufacture of concrete

El objetivo principal de este trabajo de investigación es estudiar las propiedades del árido reciclado mixto para la fabricación de hormigón reciclado en aplicaciones no estructurales. Se ha realizado la caracterización completa de 35 muestras de áridos reciclados mixtos gruesos de distinta calidad, procedentes de 13 plantas de tratamiento diferentes de la geografía española. Se han estudiado las correlaciones que existen entre las diferentes propiedades, en particular, con la absorción de agua, el contenido de sulfatos y la composición. Se propone una clasificación de los áridos reciclados y se limita de forma indicativa el contenido de yeso para que una muestra de árido reciclado mixto cumpla con la limitación del 0,8% de los sulfatos solubles en ácido de la Instrucción EHE-08. Recycling of construction and demolition waste (CDW) has become a widespread concern in Spain for the last years, as a way to preserve natural resources and achieve a better control of waste disposal sites.Specific applications which make use of mixed recycled aggregates are of great importance, as this types of aggregates constitute the majority of the total production. Structural and non-structural concrete is one of the possible applications, being this the main goal of our study. This paper presents a study on the physical and chemical characteristics of mixed recycled aggregates which have been obtained from different CDW treatment plants of Spain. Correlations between the different properties were investigated in order to find criterions of acceptance for recycled aggregates to be used in concrete. The comparison between the properties offers the possibility of pre-selecting a great quantity of mixed recycled aggregates, these being suitable for either structural and non-structural concrete. The determination of water absorption and the gypsum content are good indicators in order to evaluate the quality of the mixed recycled aggregates for its application in the production of concrete.

Operational characterisation of a Micro-SME producing farmhouse cheese in the south of Venezuela

A piece of research is presented that was conducted on the Guayanes Farmhouse Telita Cheese Producers Network located in the Piar and Padre Chien rural municipalities of Bolivar state in Venezuela. Guayanes telita cheese is a regional dairy product. The producers are to be found in a rural area with a high potential for marketing the label in the Southern Common Market (MERCOSUR). This market is the focal point of the strategic importance of this study for the Region and the Country. The research is of a descriptive scope conducted in the field. A questionnaire based on good food production practice was used as a data gathering technique. The final sample comprised 30 production units. Statistical processing was performed with version 15.2 of the STATGRAPHICS Centurion computational tool. The results would appear to confirm previous studies that point to the existence of factors that prevent these Micro-SMEs from guaranteeing the food safety of the product. The results indicate that new lines of research need to be opened up. These are oriented towards formulating strategies for the continuous improvement of these micro-SMEs, including quality control indicators.

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.

Non-destructive optical characterisation of fruits with time-resolved reflectance spectroscopy.

Increasing attention is being paid to the possible development of non-invasive tests for the assessment of the quality of fruits We propose a novel non-destructive method for the measurement of the internal optical properties of fruits and vegetables by means of time resolved reflectance spectroscopy in the visible and NIR range. A fully automated instrumentation for time-resolved reflectance measurements was developed It is based on mode-locked laser sources and electronics for time-correlated single photon counting, and provides a time-resolution of 120-160 ps The system was used to probe the optical properties of several species and varieties of fruits and vegetables in the red and NIR range (650-1000 nm). In most fruits, the absorption line shape is dominated by the absorption peak of water, centred around 970 nm Generally, the absorption spectra also show the spectral features typical of chlorophyll, with maximum at 675 nm In particular, for what concerns apples, variations in peak intensity are observed depending on the variety, the degree of ripeness as well as the position on the apple. For all the species and varieties considered, the transport scattering coefficient decreases progressively upon increasing the wavelength.

Automated characterisation of intergrowth textures in mineral particles. A case study

The characterisation of mineral texture has been a major concern for process mineralogists, as liberation characteristics of the ores are intimately related to the mineralogical texture. While a great effort has been done to automatically characterise texture in unbroken ores, the characterisation of textural attributes in mineral particles is usually descriptive. However, the quantitative characterisation of texture in mineral particles is essential to improve and predict the performance of minerallurgical processes (i.e. all the processes involved in the liberation and separation of the mineral of interest) and to achieve a more accurate geometallurgical model. Driven by this necessity of achieving a more complete characterisation of textural attributes in mineral particles, a methodology has been recently developed to automatically characterise the type of intergrowth between mineral phases within particles by means of digital image analysis. In this methodology, a set ofminerallurgical indices has been developed to quantify different mineralogical features and to identify the intergrowth pattern by discriminant analysis. The paper shows the application of the methodology to the textural characterisation of chalcopyrite in the rougher concentrate of the Kansanshi copper mine (Zambia). In this sample, the variety of intergrowth patterns of chalcopyrite with the other minerals has been used to illustrate the methodology. The results obtained show that the method identifies the intergrowth type and provides quantitative information to achieve a complete and detailed mineralogical characterisation. Therefore, the use of this methodology as a routinely tool in automated mineralogy would contribute to a better understanding of the ore behaviour during liberation and separation processes.

Characterisation of laser hybrid welded dissimilar joints

The investigation addresses the over-all performance of dissimilar joints of low carbon steel and stainless steel thin sheets achieved by laser hybrid welding. First, the technological de-velopment of dissimilar laser hybrid welding of thin sheets is briefly pre-sented. Joint characterisation by means of macro and microstructural examination and hardness tests is fur-ther described. Microhardness testing was used as an alternative and effi-cient mean of assessing the changes in mechanical properties of difficult to characterize areas, like HAZ and fu-sion zone of these thin sheets Laser-GMA dissimilar welded joints. The overall tensile performance of the joint is discussed together with the weld metal strength overmatching. The ten-sile tests results indicate that in case of transversally loaded joints, the po-sitive difference in yield strength between the weld metal and the base materials (overmatching welds) may reduce the weight of the structure, without diminishing its strength.

Optical characterisation of semi-transparent PV modules for building integration

A complete characterisation of PV modules for building integration is needed in order to know their influence on the building’s global energy balance. Specifically, certain characteristic parameters should be obtained for each different PV module suitable for building integrated photovoltaics (BIPV), some by direct or indirect measurements at the laboratory, and others by monitoring the element performance mounted in real operating conditions. In the case of transparent building envelopes it is particularly important to perform an optical and thermal characterization of the PV modules that would be integrated in them. This paper addresses the optical characterization of some commercial thin-film PV modules having different degrees of transparency, suitable for building integration in façades. The approach is based on the measurement of the spectral UV/Vis/NIR reflectance and transmittance of the different considered samples, both at normal incidence and as a function of the angle of incidence. With the obtained results, the total and zoned UV, visible and NIR transmission and reflection values are calculated, enabling the correct characterization of the PV modules integrated in façades and the subsequent evaluation of their impact over the electrical, thermal and lighting performance in a building.

Mechanical characterisation of tungsten-1wt.% yttrium oxide as a function of temperature and atmosphere

This study evaluates the mechanical behaviour of an Y2O3-dispersed tungsten (W) alloy and compares it to a pure W reference material. Both materials were processed via mechanical alloying (MA) and subsequent hot isostatic pressing (HIP). We performed non-standard three-point bending (TPB) tests in both an oxidising atmosphere and vacuum across a temperature range from 77 K, obtained via immersion in liquid nitrogen, to 1473 K to determine the mechanical strength, yield strength and fracture toughness. This research aims to evaluate how the mechanical behaviour of the alloy is affected by oxides formed within the material at high temperatures, primarily from 873 K, when the materials undergo a massive thermal degradation. The results indicate that the alloy is brittle to a high temperature (1473 K) under both atmospheres and that the mechanical properties degrade significantly above 873 K. We also used Vickers microhardness tests and the dynamic modulus by impulse excitation technique (IET) to determine the elastic modulus at room temperature. Moreover, we performed nanoindentation tests to determine the effect of size on the hardness and elastic modulus; however, no significant differences were found. Additionally, we calculated the relative density of the samples to assess the porosity of the alloy. Finally, we analysed the microstructure and fracture surfaces of the tested materials via field emission scanning electron microscopy (FE-SEM) and transmission electron microscopy (TEM). In this way, the relationship between the macroscopic mechanical properties and micromechanisms of failure could be determined based on the temperature and oxides formed

Short communication. Collection and characterisation of a population of Triticum boeoticum Boiss., a wild wheat species not previously found in the Mediterranean western region

En julio del 2010 se recolectó una población de trigo silvestre en una zona abandonada cerca de Madrid, España. Esta zona posee una biodiversidad botánica elevada y un tipo de suelo muy peculiar denominado “arcillas verdes”. Se recogió una muestra de trigo y se multiplicó y caracterizó para varios caracteres agro-morfológicos y subunidades de gluteninas. El número cromosómico 2n de las semillas demostró que es una especie diploide de trigo y los datos de caracterización indicaron que es Triticum boeoticum Boiss. Esta especie llegó probablemente como mala hierba del cultivo de escaña que se producía en la zona hasta al menos la primera mitad del s. xix. Las características edáficas y climáticas del lugar y el hecho de que no haya referencias hasta ahora de esta especie en la zona oeste de la región Mediterránea aumentan el valor de esta adquisición para la mejora del trigo. La nueva accesión se conserva en el Centro Nacional de Recursos Fitogenéticos y se ha depositado una hoja de herbario en el Real Jardín Botánico de Madrid

Advanced Characterisation of a Coffee Fermenting Tank by Multi-distributed Wireless Sensors: Spatial Interpolation and Phase Space Graphs

The fermentation stage is considered to be one of the critical steps in coffee processing due to its impact on the final quality of the product. The objective of this work is to characterise the temperature gradients in a fermentation tank by multi-distributed, low-cost and autonomous wireless sensors (23 semi-passive TurboTag® radio-frequency identifier (RFID) temperature loggers). Spatial interpolation in polar coordinates and an innovative methodology based on phase space diagrams are used. A real coffee fermentation process was supervised in the Cauca region (Colombia) with sensors submerged directly in the fermenting mass, leading to a 4.6 °C temperature range within the fermentation process. Spatial interpolation shows a maximum instant radial temperature gradient of 0.1 °C/cm from the centre to the perimeter of the tank and a vertical temperature gradient of 0.25 °C/cm for sensors with equal polar coordinates. The combination of spatial interpolation and phase space graphs consistently enables the identification of five local behaviours during fermentation (hot and cold spots).