The Size of a Graph Without Topological Complete Subgraphs

In this note we show a new upperbound for the function ex(n;TKp), i.e., the maximum number of edges of a graph of order n not containing a subgraph homeomorphic to the complete graph of order p. Further, for ${\left \lceil \frac{2n+5}{3}\right \rceil}\leq p < n$ we provide exact values for this function

The center of an infinite graph

In this note we extend the notion of the center of a graph to infinite graphs. Thus, a vertex is in the center of the infinite graph G if it is in the center of an increasing family of finite subgraphs covering G. We give different characterizations of when a vertex is in the center of an infinite graph and we prove that any infinite graph with at least two ends has a center

Extremal Graphs without Topological Complete Subgraphs

The exact values of the function $ex(n;TK_{p})$ are known for ${\lceil \frac{2n+5}{3}\rceil}\leq p < n$ (see [Cera, Diánez, and Márquez, SIAM J. Discrete Math., 13 (2000), pp. 295--301]), where $ex(n;TK_p)$ is the maximum number of edges of a graph of order n not containing a subgraph homeomorphic to the complete graph of order $p.$ In this paper, for ${\lceil \frac{2n+6}{3} \rceil}\leq p < n - 3,$ we characterize the family of extremal graphs $EX(n;TK_{p}),$ i.e., the family of graphs with n vertices and $ex(n;TK_{p})$ edges not containing a subgraph homeomorphic to the complete graph of order $p.

Attributions for success and failure in Spanish team sport players

[EN] This study examined the role of ability, effort, luck and difficulty of the task in attributions for successful and unsuccessful performances made by Spanish team sport players, and determined if differences exist between athletes at different competition levels. Participants were 143 young men (soccer: N = 64; indoor soccer: N = 37; basketball: N = 42), ages 17 to 25 years (M = 20, SD = 5). Winners perceived ability and effort as the reasons behind their success while losers made attributions mainly to luck and the difficulty of the task. Ability and effort were in the three sports the reasons given for successful outcomes at various competition levels. Difficulty of the task was the factor rated higher by those who lost competing at a national level, while luck and effort were rated higher for those who lost competing at a regional and local level.[ES] En este estudio se analizaron el papel de la habilidad, el esfuerzo, la suerte y la dificultad de la tarea en las atribuciones de éxito y fracaso realizadas por practicantes españoles de deportes colectivos y se determinó si existían diferencias entre jugadores con distintos niveles de competición. Participaron 143 chicos (fútbol: N = 64; fútbol sala: N = 37; baloncesto: N = 42), con edades comprendidas entre 17 y 25 años (M = 20, SD = 5). Los ganadores consideraban la habilidad y el esfuerzo como las razones responsables de su éxito, mientras que los perdedores realizaban sus atribuciones principalmente a la suerte y a la dificultad de la tarea. En los tres deportes estudiados, la habilidad y el esfuerzo se consideraban como las principales razones del éxito en los diferentes niveles competitivos. La dificultad de la tarea era la causa más puntuada por los perdedores que competían a nivel nacional, mientras que a nivel regional y local se otorgaba mayor importancia a la suerte y al esfuerzo.S

Production of highly bioactive resveratrol analogues pterostilbene and piceatannol in metabolically engineered grapevine cell cultures

Summary Grapevine stilbenes, particularly trans-resveratrol, have a demonstrated pharmacological activity. Other natural stilbenes derived from resveratrol such as pterostilbene or piceatannol, display higher oral bioavailability and bioactivity than the parent compound, but are far less abundant in natural sources. Thus, to efficiently obtain these bioactive resveratrol derivatives, there is a need to develop new bioproduction systems. Grapevine cell cultures are able to produce large amounts of easily recoverable extracellular resveratrol when elicited with methylated cyclodextrins and methyl jasmonate. We devised this system as an interesting starting point of a metabolic engineering-based strategy to produce resveratrol derivatives using resveratrolconverting enzymes. Constitutive expression of either Vitis vinifera resveratrol O-methyltransferase (VvROMT) or human cytochrome P450 hydroxylase 1B1 (HsCYP1B1) led to pterostilbene or piceatannol, respectively, after the engineered cell cultures were treated with the aforementioned elicitors. Functionality of both gene products was first assessed in planta by Nicotiana benthamiana agroinfiltration assays, in which tobacco cells transiently expressed stilbene synthase and VvROMT or HsCYP1B1. Grapevine cell cultures transformed with VvROMT produced pterostilbene, which was detected in both intra- and extracellular compartments, at a level of micrograms per litre. Grapevine cell cultures transformed with HsCYP1B1 produced about 20 mg/L culture of piceatannol, displaying a sevenfold increase in relation to wild-type cultures, and reaching an extracellular distribution of up to 45% of total production. The results obtained demonstrate the feasibility of this novel system for the bioproduction of natural and more bioactive resveratrol derivatives and suggest new ways for the improvement of production yield

A database and digital signal processing framework for the perceptual analysis of voice quality

Bermúdez de Alvear RM, Corral J, Tardón LJ, Barbancho AM, Fernández Contreras E, Rando Márquez S, Martínez-Arquero AG, Barbancho I. A database and digital signal processing framework for the perceptual analysis of voice quality. Pan European Voice Conferenc: PEVOC 11 Abstract Book. Aug. 31-Sept.2, 2015.Introduction. Clinical assessment of dysphonia relies on perceptual as much as instrumental methods of analysis [1]. The perceptual auditory analysis is potentially subject to several internal and external sources of bias [2]. Furthermore acoustic analyses which have been used to objectively characterize pathological voices are likely to be affected by confusion variables such as the signal processing or the hardware and software specifications [3]. For these reasons the poor correlation between perceptual ratings and acoustic measures remains to be a controversial matter [4]. The availability of annotated databases of voice samples is therefore of main importance for clinical and research purposes. Databases to perform digital processing of the vocal signal are usually built from English speaking subjects’ sustained vowels [5]. However phonemes vary from one language to another and to the best of our knowledge there are no annotated databases with Spanish sustained vowels from healthy or dysphonic voices. This work shows our first steps to fill in this gap. For the aim of aiding clinicians and researchers in the perceptual assessment of voice quality a two-fold objective was attained. On the one hand a database of healthy and disordered Spanish voices was developed; on the other an automatic analysis scheme was accomplished on the basis of signal processing algorithms and supervised learning machine techniques. Material and methods. A preliminary annotated database was created with 119 recordings of the sustained Spanish /a/; they were perceptually labeled by three experienced experts in vocal quality analysis. It is freely available under Links in the ATIC website (www.atic.uma.es). Voice signals were recorded using a headset condenser cardioid microphone (AKG C-544 L) positioned at 5 cm from the speaker’s mouth commissure. Speakers were instructed to sustain the Spanish vowel /a/ for 4 seconds. The microphone was connected to a digital recorder Edirol R-09HR. Voice signals were digitized at 16 bits with 44100 Hz sampling rate. Afterwards the initial and last 0.5 second segments were cut and the 3 sec. mid portion was selected for acoustic analysis. Sennheiser HD219 headphones were used by judges to perceptually evaluate voice samples. To label these recordings raters used the Grade-Roughness-Breathiness (GRB) perceptual scale which is a modified version of the original Hirano’s GRBAS scale, posteriorly modified by Dejonckere et al., [6]. In order to improve intra- and inter-raters’ agreement two types of modifications were introduced in the rating procedure, i.e. the 0-3 points scale resolution was increased by adding subintervals to the standard 0-3 intervals, and judges were provided with a written protocol with explicit definitions about the subintervals boundaries. By this way judges could compensate for the potential instability that might occur in their internal representations due to the perceptual context influence [7]. Raters’ perceptual evaluations were simultaneously performed by means of connecting the Sennheiser HD219 headphones to a multi-channel headphone preamp Behringer HA4700 Powerplay Pro-XL. The Yin algorithm [8] was selected as initial front-end to identify voiced frames and extract their fundamental frequency. For the digital processing of voice signals some conventional acoustic parameters [6] were selected. To complete the analysis the Mel-Frequency Cepstral Coefficients (MFCC) were further calculated because they are based on the auditory model and they are thus closer to the auditory system response than conventional features. Results. In the perceptual evaluation excellent intra-raters agreement and very good inter-raters agreement were achieved. During the supervised machine learning stage some conventional features were found to attain unexpected low performance in the classification scheme selected. Mel Frequency Cepstral Coefficients were promising for assorting samples with normal or quasi-normal voice quality. Discussion and conclusions. Despite it is still small and unbalanced the present annotated data base of voice samples can provide a basis for the development of other databases and automatic classification tools. Other authors [9, 10, 11] also found that modeling the auditory non-linear response during signal processing can help develop objective measures that better correspond with perceptual data. However highly disordered voices classification remains to be a challenge for this set of features since they cannot be correctly assorted by either conventional variables or the auditory model based measures. Current results warrant further research in order to find out the usability of other types of voice samples and features for the automatic classification schemes. Different digital processing steps could be used to improve the classifiers performance. Additionally other types of classifiers could be taken into account in future studies. Acknowledgment. This work was funded by the Spanish Ministerio de Economía y Competitividad, Project No. TIN2013-47276-C6-2-R has been done in the Campus de Excelencia Internacional Andalucía Tech, Universidad de Málaga. References [1] Carding PN, Wilson JA, MacKenzie K, Deary IJ. Measuring voice outcomes: state of the science review. The Journal of Laryngology and Otology 2009;123,8:823-829. [2] Oates J. Auditory-perceptual evaluation of disordered voice quality: pros, cons and future directions. Folia Phoniatrica et Logopaedica 2009;61,1:49-56. [3] Maryn et al. Meta-analysis on acoustic voice quality measures. J Acoust Soc Am 2009; 126, 5: 2619-2634. [4] Vaz Freitas et al. Correlation Between Acoustic and Audio-Perceptual Measures. J Voice 2015;29,3:390.e1 [5] “Multi-Dimensional Voice Program (MDVP) Model 5105. Software Instruction Manual”, Kay PENTAX, A Division of PENTAX Medical Company, 2 Bridgewater Lane, Lincoln Park, NJ 07035-1488 USA, November 2007. [6] Dejonckere PH, Bradley P, Clemente P, Cornut G, Crevier-Buchman L, Friedrich G, Van De Heyning P, Remacle M, Woisard V. A basic protocol for functional assessment of voice pathology, especially for investigating the efficacy of (phonosurgical) treatments and evaluating new assessment techniques. Guideline elaborated by the Comm. on Phoniatrics of the European Laryngological Society (ELS). Eur Arch Otorhinolaryngol 2001;258:77–82. [7] Kreiman et al. Voice Quality Perception. J Speech Hear Res 1993;36:21-4 [8] De Cheveigné A, Kawahara H. YIN, a fundamental frequency estimator for speech and music. J. Acoust. Soc. Amer. 202; 111,4:1917. [9] Shrivastav et al. Measuring breathiness. J Acoust Soc Am 2003;114,4:2217-2224. [10] Saenz-Lechon et al. Automatic Assessment of voice quality according to the GRBAS scale. Eng Med Biol Soc Ann 2006;1:2478-2481. [11] Fredouille et al. Back-and-forth methodology for objective voice quality assessment: from/to expert knowledge to/from automatic classification of dysphonia. EURASIP J Appl Si Pr 2009.Campus de Excelencia Internacional Andalucía Tech, Universidad de Málaga. Ministerio de Economía y Competitividad, Projecto No. TIN2013-47276-C6-2-R

Controlled manipulation of enzyme specificity through immobilization-induced flexibility constraints

It is thought that during immobilization enzymes, as dynamic biomolecules, may become distorted and this may alter their catalytic properties. However, the effects of different immobilization strategies on enzyme rigidity or flexibility and their consequences in specificity and stereochemistry at large scale has not been yet clearly evaluated and understood. This was here investigated by using as model an ester hydrolase, isolated from a bacterium inhabiting a karstic lake, with broad substrate spectrum (72 esters being converted; 61.5 U mg−1 for glyceryl tripropionate) but initially non-enantiospecific. We found that the enzyme (7 nm × 4.4 nm × 4.2 nm) could be efficiently ionic exchanged inside the pores (9.3 nm under dry conditions) of amino-functionalized ordered mesoporous material (NH2-SBA-15), achieving a protein load of 48 mg g−1, and a specific activity of 4.5 ± 0.1 U mg−1. When the enzyme was site-directed immobilized through His interaction with an immobilized cationon the surface of two types of magnetic micro-particles through hexahistidine-tags, protein loads up to 10.2 μg g−1 and specific activities of up to 29.9 ± 0.3 U mg−1, were obtained. We found that ionically exchanged enzyme inside pores of NH2-SBA-15 drastically narrowed the substrate range (17 esters), to an extent much higher than ionically exchanged enzyme on the surface of magnetic micro-particles (up to 61 esters). This is attributed to differences in surface chemistry, particle size, and substrate accessibility to the active site tunnel. Our results also suggested, for the first time, that immobilization of enzymes in pores of similar size may alter the enzyme structures and produce enzyme active centers with different configuration which promote stereochemical conversions in a manner different to those arising from surface immobilization, where the strength of the ionic exchange also has an influence. This was shown by demonstrating that when the enzyme was introduced inside pores with a diameter (under dry conditions) slightly higher than that of the enzyme crystal structure a biocatalyst enantiospecific for ethyl (R)-4-chloro-3-hydroxybutyrate was produced, a feature not found when using wider pores. By contrast, immobilization on the surface of ferromagnetic microparticles produced selective biocatalysts for methyl (S)-(+)-mandelate or methyl (S)-lactate depending on the functionalization. This study illustrates the benefits of extensive analysis of the substrate spectra to better understand the effects of different immobilization strategies on enzyme flexibility/rigidity, as well as substrate specificity and stereochemistry. Our results will help to design tunable materials and interfaces for a controlled manipulation of specificity and to transform non-enantiospecific enzymes into stereo-chemically substrate promiscuous biocatalysts capable of converting multiple chiral molecules.This project received funding from the European Union’s Horizon 2020 research and innovation program Blue Growth: Unlockingthe potential of Seas and Oceans under grant agreement no. 634486 (project acronym INMARE). This research was also supported by the grants PCIN-2014-107 (within ERA NET IB2 grant nr. ERA-IB-14-030 - MetaCat), PCIN-2017-078 (within the ERA-MarineBiotech grant ProBone), BIO2014-54494-R, MAT2016-77496-R, BIO2017-85522-R, and CTQ2016-79138-R from the Spanish Ministry of Economy, Industry and Competitiveness. A.B. acknowledges the support from the Spanish Ministry of Economy, Industry and Competitiveness (MAT2017-88808-R grant), María de Maeztu Units of Excellence Programme (MDM-2016-0618), and the Diputación de Guipúzcoa for current funding in the frame of Gipuzkoa Fellows program. G.D. thanks the German Federal Ministry of Education and Research (BMBF, Grant No. 031A095C) for funding in the frame of the Molecular Interaction Engineering program (Biotechnologie 2020+). The authors gratefully acknowledge financial support provided by the European Regional Development Fund (ERDF). C.C. thanks the Spanish Ministry of Economy, Industry and Competitiveness for a PhD fellowship (Grant BES-2015-073829).Peer ReviewedPostprint (published version

Llegint un àlbum il·lustrat per expressar el model Terra

Aquesta experiència es basa en la reflexió sobre les idees que poden emergir del model Terra a partir de la lectura de l'àlbum il·lustrat "En Jan i en Pep fan un forat" amb alumnes de primària. L'estratègia de confrontar les idees dels infants des de la realitat i la ficció ens ha permès indagar en el paper de les representacions a l'hora d'estructurar idees científique

Production of highly bioactive resveratrol analogues pterostilbene and piceatannol in metabolically engineered grapevine cell cultures

Grapevine stilbenes, particularly trans-resveratrol, have a demonstrated pharmacological activity. Other natural stilbenes derived from resveratrol such as pterostilbene or piceatannol, display higher oral bioavailability and bioactivity than the parent compound, but are far less abundant in natural sources. Thus, to efficiently obtain these bioactive resveratrol derivatives, there is a need to develop new bioproduction systems. Grapevine cell cultures are able to produce large amounts of easily recoverable extracellular resveratrol when elicited with methylated cyclodextrins and methyl jasmonate. We devised this system as an interesting starting point of a metabolic engineering-based strategy to produce resveratrol derivatives using resveratrol-converting enzymes. Constitutive expression of either Vitis vinifera resveratrol O-methyltransferase (VvROMT) or human cytochrome P450 hydroxylase 1B1 (HsCYP1B1) led to pterostilbene or piceatannol, respectively, after the engineered cell cultures were treated with the aforementioned elicitors. Functionality of both gene products was first assessed in planta by Nicotiana benthamiana agroinfiltration assays, in which tobacco cells transiently expressed stilbene synthase and VvROMT or HsCYP1B1. Grapevine cell cultures transformed with VvROMT produced pterostilbene, which was detected in both intra- and extracellular compartments, at a level of micrograms per litre. Grapevine cell cultures transformed with HsCYP1B1 produced about 20 mg/L culture of piceatannol, displaying a sevenfold increase in relation to wild-type cultures, and reaching an extracellular distribution of up to 45% of total production. The results obtained demonstrate the feasibility of this novel system for the bioproduction of natural and more bioactive resveratrol derivatives and suggest new ways for the improvement of production yields.This work has been supported by grants from the Spanish Ministry of Science and Innovation (BIO2011-29856-C02-01, BIO2011-29856-C02-02 and BIO2014-51861-R), Generalitat de Catalunya (2014SGR215) and European Funds for Regional Development (FEDER) and Conselleria d’Educacio, Cultura i Sport de la Generalitat Valenciana (FPA/2013/A/074). J.M.C. acknowledges a postdoctoral and research grants from SENESCYT GOVERNMENT OF ECUADOR (006-IECE-SMG5-GPLR-2012 and Programa1-Senescyt-2014) and a grant from UTEQ (UTEQAmbiental-9-FCAmb-IFOR-2014-FOCICYT002)

Activos, productos y mercados financieros: entre todos y para todos

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