Identification and characterisation of ncRNAs in dormant bacteria

Recent years have led to increasing interest and appreciation of the possible importance of single cell heterogeneity in various biological processes. One of the examples of phenotypic heterogeneity in bacterial populations is antibiotic tolerant persister cells. Such an antibiotic tolerance phenotype is of considerable clinical relevance since dormant bacteria can re-establish infections rapidly after the antibiotic treatment has been terminated. Up to now mechanisms for establishing the persistence phenomenon in bacteria have remained largely enigmatic. Persisters are cells considered to be in a dormant state with down regulated gene expression. Only recently small regulatory RNAs (sRNAs) have been appreciated as important regulators of gene expression in response to environmental stimuli and several theoretical studies have suggested a possible involvement of sRNAs in the mechanisms of regulated heterogeneity in bacteria. We have experimentally addressed this potential link between sRNAs and persistence/dormancy in E. coli as an example of heterogeneity. Beside classical sRNAs we are focusing also on sRNAs directly associating with and possibly regulating the ribosome, the central enzyme of gene expression. The persister and dormant cell specific sRNA profile is studied by the comparative analysis of sRNA profile changes of the whole bacterial population after antibiotic killing. From RNA-Seq data ~ 25 000 potentially stable RNA fragments were identified and initial analysis predicted ~300 of them to be dormant/persister cell specific. After further evaluation the most prominent dormant/persister cell specific sRNAs are functionally characterized and their potential role in the persistence/dormancy will be evaluated by applying genetic, molecular and biochemical tools. The potential results of this project will provide a better understanding on the molecular mechanism of bacterial persistence/dormancy and on the role of ribosome-bound sRNA molecules in fine-tuning gene expression.

Identification and characterisation of ncRNAs in dormant bacteria

Recent years have led to increasing interest and appreciation of the possible importance of single cell heterogeneity in various biological processes. One of the examples of phenotypic heterogeneity in bacterial populations is antibiotic tolerant persister cells. Such an antibiotic tolerance phenotype is of considerable clinical relevance since dormant bacteria can re-establish infections rapidly after the antibiotic treatment has been terminated. Up to now mechanisms for establishing the persistence phenomenon in bacteria have remained largely enigmatic. Persisters are cells considered to be in a dormant state with down regulated gene expression. Only recently small regulatory RNAs (sRNAs) have been appreciated as important regulators of gene expression in response to environmental stimuli and several theoretical studies have suggested a possible involvement of sRNAs in the mechanisms of regulated heterogeneity in bacteria. We have experimentally addressed this potential link between sRNAs and persistence/dormancy in E. coli as an example of heterogeneity. Beside classical sRNAs we are focusing also on sRNAs directly associating with and possibly regulating the ribosome, the central enzyme of gene expression. The persister and dormant cell specific sRNA profile is studied by the comparative analysis of sRNA profile changes of the whole bacterial population after antibiotic killing. From RNA-Seq data ~ 25 000 potentially stable RNA fragments were identified and initial analysis predicted ~300 of them to be dormant/persister cell specific. After further evaluation the most prominent dormant/persister cell specific sRNAs are functionally characterized and their potential role in the persistence/dormancy will be evaluated by applying genetic, molecular and biochemical tools. The potential results of this project will provide a better understanding on the molecular mechanism of bacterial persistence/dormancy and on the role of ribosome-bound sRNA molecules in fine-tuning gene expression.

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.

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.

Characterisation factors for life cycle impact assessment of sound emissions

abstract to be added by authors

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.

Image characterisation based on the mammalian visual cortex

A proposal for a model of the primary visual cortex is reported. It is structured with the basis of a simple unit cell able to perform fourteen pairs of different boolean functions corresponding to the two possible inputs. As a first step, a model of the retina is presented. Different types of responses, according to the different possibilities of interconnecting the building blocks, have been obtained. These responses constitute the basis for an initial configuration of the mammalian primary visual cortex. Some qualitative functions, as symmetry or size of an optical input, have been obtained. A proposal to extend this model to some higher functions, concludes the paper.

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.