Physiological features of Schizosaccharomyces pombe of interest in the making of white wines

This work studies the physiology of Schizosaccharomyces pombe strain 938 in the production of white wine with high malic acid levels as the sole fermentative yeast, as well as in mixed and sequential fermentations with Saccharomyces cerevisiae Cru Blanc. The induction of controlled maloalcoholic fermentation through the use of Schizosaccharomyces spp. is now being viewed with much interest. The acetic, malic and pyruvic acid concentrations, relative density and pH of the musts were measured over the entire fermentation period. In all fermentations in which Schizo. pombe 938 was involved, nearly all the malic acid was consumed and moderate acetic concentrations produced. The urea content and alcohol level of these wines were notably lower than in those made with Sacch. cerevisiae Cru Blanc alone. The pyruvic acid concentration was significantly higher in Schizo. pombe fermentations. The sensorial properties of the different final wines varied widely.

Influence of some oenological practices on the aromatic and sensorial characteristics of white Verdejo wines

Changes in the aromatic composition as well as sensory characteristics in Verdejo white wines were analysed based on two factors: the winemaking methodology and the storing time of wine in bottles. The volatile components were determined by GLC-MS, and the sensory profile was designed and assessed according to the ISO 11035 standard. The results showed that when wines were made in oak barrels, either completely or partially, which means the wines were in contact with the lees, the levels of 1-octanol, ethyl heptanoate and ethyl decanoate were significantly affected (P menor que 0.05); the softness sensation was also influenced (P menor que 0.05). However, the amount of time the wines were stored in bottles significantly affected (P menor que 0.05) the levels of 1-hexanol, ethyl heptanoate, ethyl octanoate, ethyl decanoate, hexyl acetate, isoamyl acetate and isoamyl lactate and also an odour note (tropical fruit). The compounds with higher OAV values belong to the groups of esters and fatty acids. For these reasons, the composition and the quality of the aroma of Verdejo white wines appear to be significantly affected both by use of oak barrels in winemaking and the time the wines are stored in bottles.

In vitro reconstitution of light-harvesting POR-protochlorophyllide complex with protochlorophyllides a and b

NADPH:protochlorophyllide oxidoreductase (POR; EC1.1.33.1) is a key enzyme for the light-induced greening of angiosperms. In barley, two POR proteins exist, termed PORA and PORB. These have previously been proposed to form higher molecular weight light-harvesting complexes in the prolamellar body of etioplasts (Reinbothe, C., Lebedev, N., and Reinbothe, S. (1999)Nature 397, 80–84). Here we report the in vitro reconstitution of such complexes from chemically synthesized protochlorophyllides (Pchlides) a andb and galacto- and sulfolipids. Low temperature (77 K) fluorescence measurements revealed that the reconstituted, lipid-containing complex displayed the same characteristics of photoactive Pchlide 650/657 as the presumed native complex in the prolamellar body. Moreover, Pchlide F650/657 was converted to chlorophyllide (Chlide) 684/690 upon illumination of the reconstituted complex with a 1-ms flash of white light. Identification and quantification of acetone-extractable pigments revealed that only the PORB-bound Pchlide a had been photoactive and was converted to Chlide a, whereas Pchlide b bound to the PORA remained photoinactive. Nondenaturing PAGE of the reconstituted Pchlide a/b-containing complex further demonstrated a size similar to that of the presumed native complexin vivo, suggesting that both complexes may be identical.

New data on the Pleistocene of Trani (Adriatic Coast, Southern Italy)

Along the Apulian Adriatic coast, in a cliff south of Trani, a succession of three units (superimposed on one another) of marine and/or paralic environments has been recognised. The lowest unit I is characterised by calcareous/siliciclastic sands (css), micritic limestones (ml), stromatolitic and characean boundstones (scb), characean calcarenites (cc). The sedimentary environment merges from shallow marine, with low energy and temporary episodes of subaerial exposure, to lagoonal with a few exchanges with the sea. The lagoonal stromatolites (scb subunit) grew during a long period of relative stability of a high sea level in tropical climate. The unit I is truncated at the top by an erosion surface on which the unit II overlies; this consists of a basal pebble lag (bpl), silicicla - stic sands (ss), calcareous sands (cs), characean boundstones (cb), brown paleosol (bp). The sedimentary environment varies from beach to lagoon with salinity variations. Although there are indications of seismic events within the subunits cs, unit II deposition took place in a context of relative stability. The unit II is referable to a sea level highstand. Unit III, trangressive on the preceding, consists of white calcareous sands (wcs), calcareous sands and calcarenites (csc), phytoclastic calcirudite and phytohermal travertine (pcpt), mixed deposits (csl, m, k, c), sands (s) and red/brown paleosols (rbp). The sedimentation of this unit was affected by synsedimentary tectonic, attested by seismites found at several heights. Also the unit III is referable to a sea level highstand. The scientific literature has so far generally attributed to the Tyrrhenian (auct.) the deposits of Trani cliff. As part of this work some datings were performed on 10 samples, using the amino acid racemization method (AAR) applied to ostracod carapaces. Four of these samples have been rejected because they have shown in laboratory recent contamination. The numerical ages indicate that the deposits of the Trani cliff are older than MIS 5. The upper part of the unit I has been dated to 355±85 ka BP, thus allowing to assign the lowest stromatolitic subunit (scb) at the MIS 11 peak and the top of the unit I at the MIS 11-MIS 10 interval. The base of the unit II has been dated to 333±118 ka BP, thus attributing the erosion surface that bounds the units I and II to the MIS 10 lowstand and the lower part of the unit II to MIS 9.3. The upper part of the unit II has been dated to 234±35 ka BP, while three other numerical ages come from unit III: 303±35, 267±51, 247±61 ka BP. At present, the numerical ages cannot distinguish the sedimentation ages of units II and III, which are both related to the MIS 9.3- MIS 7.1 time range. However, the position of the units, superimposed one another, and their respective age, allows us to recognise a subsidence phase between MIS 11 and MIS 7, followed by an uplift phase between the MIS 7 and the present day, which led the deposits in their current position. This tectonic pattern is not in full agreement with what is described in the literature for the Apulian foreland.

White matter damage disorganizes brain functional networks in amnestic mild cognitive impairment

Although progressive functional brain network disruption has been one of the hallmarks of Alzheimer?s Dis- ease, little is known about the origin of this functional impairment that underlies cognitive symptoms. We in- vestigated how the loss of white matter (WM) integrity disrupts the organization of the functional networks at different frequency bands. The analyses were performed in a sample of healthy elders and mild cognitive im- pairment (MCI) subjects. Spontaneous brain magnetic activity (measured with magnetoencephalography) was characterized with phase synchronization analysis, and graph theory was applied to the functional networks. We identified WM areas (using diffusion weighted magnetic resonance imaging) that showed a statistical de- pendence between the fractional anisotropy and the graph metrics. These regions are part of an episodic mem- ory network and were also related to cognitive functions. Our data support the hypothesis that disruption of the anatomical networks influences the organization at the functional level resulting in the prodromal dementia syndrome of MCI.

Influence of castration on growth performance and carcass and meat quality of heavy female pigs

The main objetive of this Doctoral Thesis was to study the influence of female castration and pig sex on growth performance and carcass and meat quality of white pigs slaughtered at different final weights. Three experiments (Exp.) were conducted. In Exp. 1, a total of 200 (Landrace * Large White dam x Pietrain * Large White sire) gilts of 50 ± 3 days of age (23.3 ± 1.47 kg BW) was used to investigate the effects of castration (intact females, IF vs. castrated feamles, CF) and slaughter weight (106 vs. 122 kg BW) on productive performance and carcass and meat quality. There were four experimental treatments arranged as a 2 x 2 factorial and 5 replicates of 10 pigs each per treatment. Half of the gilts were ovariectomized at 58 d of age (8 days after the beginning of the trial; 29.8 ± 1.64 kg BW) whereas the other half remained intact. Meat samples were taken at m. Longissimus thoracis at the level of the last rib and subcutaneous fat samples were taken at the tail insertion. For the entire experiment period, CF had higher BW gain (P<0.05) and backfat and m. Gluteus medius (GM) fat thickness (P<0.001) than IF. However, IF had higher loin and trimmed primal cut yields (P<0.05) than CF. Meat quality was similar for IF and CF but the proportion of linoleic acid in subcutaneous fat was higher (P<0.001) for IF. Pigs slaughtered at 122 kg BW had higher (P<0.001) feed intake and poorer feed efficiency than pigs slaughtered at 106 kg BW. An increase in slaughter weight (SW) improved (P<0.001) carcass yield but decreased (P<0.05) trimmed primal cut yield. Meat from females slaughtered at the heavier BW was redder (a*; P<0.001) and had more (P<0.01) intramuscular fat and less thawing (P<0.05) and cooking (P<0.10) loss than meat from females slaughtered at the lighter BW. Also, females slaughtered at 122 kg BW had less (P<0.01) linoleic acid content in the subcutaneous fat than pigs slaughtered at 106 kg BW. Castration of gilts and slaughtering at heavier BW might be useful practices for the production of heavy pigs destined to the dry cured industry in which a certain amount of fat in the carcass is required. In contrast, when the carcasses are destined to fresh meat production, IF slaughtered at 106 kg BW are a more efficient alternative. In Exp. 2, crossbred pigs (n=240) from Pietrain*Large White sires mated to Landrace*Large White dams with an average of 100 d of age (60.5 ± 2.3 kg) were used to investigate the effects of gender and slaughter weight (SW) on growth performance and carcass and meat quality characteristics. There were 6 treatments arranged factorially with 3 genders (IF vs. CF vs.castrated males, CM) and 2 SW (114 vs. 122 kg BW). Each of the 6 combinations of treatments was replicated 4 times and the experimental unit was a pen with 10 pigs. Castrated males and CF ate more feed, grew faster and had more carcass backfat depth and fat thickness at the GM muscle, but lower loin yield than IF (P<0.05). In addition, CF and CM had more intramuscular fat (P<0.05) and less linoleic acid content in the subcutaneous fat (P<0.01) than IF. Pigs slaughtered at 122 kg BW had lower ADG (P<0.05), poor gain-to-feed ratio (P<0.05), and more GM fat than pigs slaughtered at 114 kg BW (P < 0.05). It is concluded that CF and CM had similar productive performance and meat quality characteristics when slaughtered at the same age, and that the castration of females improved daily gains and increased weight and fat content of primal cuts with respect to IF. Therefore, castration of females is recommended in pigs destined to the dry-cured industry because of the beneficial effects on the quality of the primal cuts. In Exp. 3, the effects of gender and castration of females (IF vs. CF vs. CM) on performance and carcass and meat quality were studied in crossbred pigs (Landrace x Large White dams x Duroc sires) slaughtered at 119.2 (trial 1) or 131.6 (trial 2) kg BW. Intact females had better feed conversion and less carcass fat than CF and CM. Trimmed shoulder yield was higher for CM than for CF with IF being intermediate. Primal cut yield and meat quality, however were similar for all treatments. Proportion of linoleic acid in backfat was lower for CF than for IF or CM, and the differences were significant in pigs slaughtered witn 131.6 kg BW. The higher fat content and the fatty acid profile favour the use of CF and CM over IF for the production of heavy pigs destined to the dry-cured industry.

Light regulates motility, attachment and virulence in the plant pathogen Pseudomonas syringae pv tomato DC3000.

Pseudomonas syringae pv tomato DC3000 (Pto) is the causal agent of the bacterial speck of tomato, which leads to significant economic losses in this crop. Pto inhabits the tomato phyllosphere, where the pathogen is highly exposed to light, among other environmental factors. Light represents a stressful condition and acts as a source of information associated with different plant defence levels. Here, we analysed the presence of both blue and red light photoreceptors in a group of Pseudomonas. In addition, we studied the effect of white, blue and red light on Pto features related to epiphytic fitness. While white and blue light inhibit motility, bacterial attachment to plant leaves is promoted. Moreover, these phenotypes are altered in a blue-light receptor mutant. These light-controlled changes during the epiphytic stage cause a reduction in virulence, highlighting the relevance of motility during the entry process to the plant apoplast. This study demonstrated the key role of light perception in the Pto phenotype switching and its effect on virulence.

Nitrogen fixation by native Bradyrhizobia in symbiosis with Lupinus mariae-josephae requires a T3SS encoding a NopE-like effector

Several bradyrhizobial isolates from L. mariae-josephae root nodules [1] contain a type III secretion system (T3SS) within a cluster of about 30 genes. Among those genes, ttsI codes for the transcriptional activator of the system. Mutation of ttsI resulted in the formation of white, non-fixing nodules with the natural legume host, L. mariae-josephae. The T3SS cluster also contains a gene coding for a NopE-like protein. NopE proteins have been demonstrated to be effectors in the Bradyrhizobium-soybean symbiosis [2] and belong to a small group of poorly characterized proteins from plant-associated bacteria that contain one or two autocleavage motifs known as DUF1521 (Schirrmeister et al. 2011). The amino acid sequence of a NopE-like protein in the L. mariae-josephae strain LmjC contains just one autocatalytic motif. This is unlike NopE1 and NopE2 proteins secreted by the T3SS of B. japonicum, that contain two motifs [3]. The autocleavage of LmjC NopE protein was analyzed after expression in E. coli and purification. Two protein fragments of the predicted sizes appeared in the presence of Ca2+, Cu2+, Cd2+, Zn2+ and Mn2+ cations. In contrast, autocleavage did not take place in the presence of Ni2+, Co2+ or Mg2+. Site-directed mutagenesis of the DUF1521 motif in LmjC NopE abolished self-cleavage in vitro. Symbiotic competence of a NopE- mutant with the L. mariae-josephae host was not affected. Possible roles of NopE are discussed.

Native bradyrhizobia isolated from Lupinus mariae-josephae possess an essential T3SS for symbiosis

Analysis of the genome sequence of bradyrhizobia strains isolated from root nodules of Lupinus mariae-josephae revealed the presence of a type III secretion system (T3SS). Mutagenesis of ttsI gene that codes for the transcriptional activator (TtsI) resulted in the formation of white, non-fixing nodules in L. mariae-josephae. The T3SS cluster includes a gene coding for a NopE-like protein with an autocleavage motif. The NopE protein is an effector in the Bradyrhizobium-soybean symbiosis (Wenzel et al., 2010). The autocatalytic properties of the purified NopE-like protein have been studied.

Noise-induced attractor annihilation in the delayed feedback logistic map

We study dynamics of the bistable logistic map with delayed feedback, under the influence of white Gaussian noise and periodic modulation applied to the variable. This system may serve as a model to describe population dynamics under finite resources in noisy environment with seasonal fluctuations. While a very small amount of noise has no effect on the global structure of the coexisting attractors in phase space, an intermediate noise totally eliminates one of the attractors. Slow periodic modulation enhances the attractor annihilation.

Diseño de un modelo para la identificación y análisis de tramos de carreteras sin accidentes : una nueva visión de la seguridad vial

El planteamiento tradicional de análisis de la accidentalidad en carretera pasa por la consideración de herramientas paliativas, como son la identificación y gestión de los puntos negros o tramos de concentración de accidentes, o preventivas, como las auditorías e inspecciones de seguridad vial. En esta tesis doctoral se presenta un planteamiento complementario a estas herramientas, desde una perspectiva novedosa: la consideración de los tramos donde no se producen accidentes; son los denominados Tramos Blancos. La tesis persigue demostrar que existen determinados parámetros del diseño de las carreteras y del tráfico que, bajo características generales similares de las vías, tienen influencia en el hecho de que se produzcan o no accidentes, adicionalmente a la exposición al riesgo, como factor principal, y a otros factores. La propia definición de los Tramos Blancos, entendidos como tramos de carreteras de longitud representativa donde no se han producido accidentes con víctimas mortales o heridos graves durante un periodo largo de tiempo, garantiza que esta situación no se produzca como consecuencia de la aleatoriedad de los accidentes, sino que pudiera deberse a una confluencia específica de determinados parámetros de la geometría de la vía y del tráfico total y de vehículos pesados. Para el desarrollo de esta investigación se han considerado la red de autopistas de peaje y las carreteras convencionales de la Red del Estado de España, que supone un total de 17.000 kilómetros, y los datos de accidentes con víctimas mortales y heridos graves en el periodo 2006-2010, ambos incluidos, en estas redes (un total de 10.000 accidentes). La red viaria objeto de análisis supone el 65% de la longitud de la Red de Carreteras del Estado, por la que circula el 33% de su tráfico; en ella se produjeron en el año 2013 el 47% de los accidentes con víctimas y el 60% de las víctimas mortales de la Red de Carreteras del Estado. Durante la investigación se ha desarrollado una base de datos de 250.130 registros y más de 3.5 millones de datos en el caso de las autopistas de peaje de la Red de Carreteras del Estado y de 935.402 registros y más de 14 millones de datos en el caso de la red convencional del Estado analizada. Tanto las autopistas de peaje como las carreteras convencionales han sido clasificadas según sus características de tráfico, de manera que se valoren vías con nivel de exposición al riesgo similar. Para cada tipología de vía, se ha definido como longitud de referencia para que un tramo se considere Tramo Blanco la longitud igual al percentil 95 de las longitudes de tramos sin accidentes con heridos graves o víctimas mortales durante el periodo 2006-2010. En el caso de las autopistas de peaje, en la tipología que ha sido considerada para la definición del modelo, esta longitud de referencia se estableció en 14.5 kilómetros, mientras que en el caso de las carreteras convencionales, se estableció en 7.75 kilómetros. Para cada uno de los tipos de vía considerados se han construido una base de datos en la que se han incluido las variables de existencia o no de Tramo Blanco, así como las variables de tráfico (intensidad media diaria total, intensidad de vehículos pesados y porcentaje de vehículos pesados ), la velocidad media y las variables de geometría (número de carriles, ancho de carril, ancho de arcén derecho e izquierdo, ancho de calzada y plataforma, radio, peralte, pendiente y visibilidad directa e inversa en los casos disponibles); como variables adicionales, se han incluido el número de accidentes con víctimas, los fallecidos y heridos graves, índices de peligrosidad, índices de mortalidad y exposición al riesgo. Los trabajos desarrollados para explicar la presencia de Tramos Blancos en la red de autopistas de peaje han permitido establecer las diferencias entre los valores medios de las variables de tráfico y diseño geométrico en Tramos Blancos respecto a tramos no blancos y comprobar que estas diferencias son significativas. Así mismo, se ha podido calibrar un modelo de regresión logística que explica parcialmente la existencia de Tramos Blancos, para rangos de tráfico inferiores a 10.000 vehículos diarios y para tráficos entre 10.000 y 15.000 vehículos diarios. Para el primer grupo (menos de 10.000 vehículos al día), las variables que han demostrado tener una mayor influencia en la existencia de Tramo Blanco son la velocidad media de circulación, el ancho de carril, el ancho de arcén izquierdo y el porcentaje de vehículos pesados. Para el segundo grupo (entre 10.000 y 15.000 vehículos al día), las variables independientes más influyentes en la existencia de Tramo Blanco han sido la velocidad de circulación, el ancho de calzada y el porcentaje de vehículos pesados. En el caso de las carreteras convencionales, los diferentes análisis realizados no han permitido identificar un modelo que consiga una buena clasificación de los Tramos Blancos. Aun así, se puede afirmar que los valores medios de las variables de intensidad de tráfico, radio, visibilidad, peralte y pendiente presentan diferencias significativas en los Tramos Blancos respecto a los no blancos, que varían en función de la intensidad de tráfico. Los resultados obtenidos deben considerarse como la conclusión de un análisis preliminar, dado que existen otros parámetros, tanto de diseño de la vía como de la circulación, el entorno, el factor humano o el vehículo que podrían tener una influencia en el hecho que se analiza, y no se han considerado por no disponer de esta información. En esta misma línea, el análisis de las circunstancias que rodean al viaje que el usuario de la vía realiza, su tipología y motivación es una fuente de información de interés de la que no se tienen datos y que permitiría mejorar el análisis de accidentalidad en general, y en particular el de esta investigación. Adicionalmente, se reconocen limitaciones en el desarrollo de esta investigación, en las que sería preciso profundizar en el futuro, reconociendo así nuevas líneas de investigación de interés. The traditional approach to road accidents analysis has been based in the use of palliative tools, such as black spot (or road sections) identification and management, or preventive tools, such as road safety audits and inspections. This thesis shows a complementary approach to the existing tools, from a new perspective: the consideration of road sections where no accidents have occurred; these are the so-called White Road Sections. The aim of this thesis is to show that there are certain design parameters and traffic characteristics which, under similar circumstances for roads, have influence in the fact that accidents occur, in addition to the main factor, which is the risk exposure, and others. White Road Sections, defined as road sections of a representative length, where no fatal accidents or accidents involving serious injured have happened during a long period of time, should not be a product of randomness of accidents; on the contrary, they might be the consequence of a confluence of specific parameters of road geometry, traffic volumes and heavy vehicles traffic volumes. For this research, the toll motorway network and single-carriageway network of the Spanish National Road Network have been considered, which is a total of 17.000 kilometers; fatal accidents and those involving serious injured from the period 2006-2010 have been considered (a total number of 10.000 accidents). The road network covered means 65% of the total length of the National Road Network, which allocates 33% of traffic volume; 47% of accidents with victims and 60% of fatalities happened in these road networks during 2013. During the research, a database of 250.130 registers and more than 3.5 million data for toll motorways and 935.042 registers and more than 14 million data for single carriageways of the National Road Network was developed. Both toll motorways and single-carriageways have been classified according to their traffic characteristics, so that the analysis is performed over roads with similar risk exposure. For each road type, a reference length for White Road Section has been defined, as the 95 percentile of all road sections lengths without accidents (with fatalities or serious injured) for 2006-2010. For toll motorways, this reference length concluded to be 14.5 kilometers, while for single-carriageways, it was defined as 7.75 kilometers. A detailed database was developed for each type of road, including the variable “existence of White Road Section”, as well as variables of traffic (average daily traffic volume, heavy vehicles average daily traffic and percentage of heavy vehicles from the total traffic volume), average speed and geometry variables (number of lanes, width of lane, width of shoulders, carriageway width, platform width, radius, superelevation, slope and visibility); additional variables, such as number of accidents with victims, number of fatalities or serious injured, risk and fatality rates and risk exposure, have also been included. Research conducted for the explanation of the presence of White Road Sections in the toll motorway network have shown statistically significant differences in the average values of variables of traffic and geometric design in White Road Sections compared with other road sections. In addition, a binary logistic model for the partial explanation of the presence of White Road Sections was developed, for traffic volumes lower than 10.000 daily vehicles and for those running from 10.000 to 15.000 daily vehicles. For the first group, the most influent variables for the presence of White Road Sections were the average speed, width of lane, width of left shoulder and percentage of heavy vehicles. For the second group, the most influent variables were found to be average speed, carriageway width and percentage of heavy vehicles. For single-carriageways, the different analysis developed did not reach a proper model for the explanation of White Road Sections. However, it can be assumed that the average values of the variables of traffic volume, radius, visibility, superelevation and slope show significant differences in White Road Sections if compared with others, which also vary with traffic volumes. Results obtained should be considered as a conclusion of a preliminary analysis, as there are other parameters, not only design-related, but also regarding traffic, environment, human factor and vehicle which could have an influence in the fact under research, but this information has not been considered in the analysis, as it was not available. In parallel, the analysis of the circumstances around the trip, including its typology and motivation is an interesting source of information, from which data are not available; the availability of this information would be useful for the improvement of accident analysis, in general, and for this research work, in particular. In addition, there are some limitations in the development of the research work; it would be necessary to develop an in-depth analysis in the future, thus assuming new research lines of interest.