Purification of Starch Granules from Arabidopsis Leaves and Determination of Granule-Bound Starch Synthase Activity

Starch constitutes the most important carbon reserve in plants and is composed of branched amylopectin and linear amylose. The latter is synthesized exclusively by the Granule-Bound Starch Synthase (GBSS, EC 2.4.1.21). Here we report a readily reproducible, specific and highly sensitive protocol, which includes the isolation of intact starch granules from Arabidopsis thaliana leaves and the subsequent determination of GBSS activity. We have applied this method to study GBSS activity in diurnal cycles in vegetative growth and during the photoperiodic transition to flowering in Arabidopsis (Tenorio et al., 2003; Ortiz-Marchena et al., 2014).España,MINECO CSD2007-00057, BIO2008-02292, and BIO2011-28847-C02-00España, Junta de Andalucía P06-CVI-01450 and P08-AGR-0358

Determination of Soluble Sugars in Arabidopsis thaliana Leaves by Anion Exchange Chromatography

Determination of soluble sugars is basic for the study of carbon metabolism in plants. Soluble sugar quantitation can be achieved by enzymatic methods implying different coupled reactions. Here we describe a simple method that allows rapid determination of the most abundant soluble sugars (glucose, fructose and sucrose) in Arabidopsis leaves by anion exchange chromatography. We have applied this method to study the levels of soluble sugars during the photoperiodic transition to flowering (Ortiz-Marchena et al., 2014).España, MINECO projects CSD2007-00057, BIO2008-02292, and BIO2011-28847-C02-00España, Junta de Andalucía P06-CVI-01450 and P08-AGR-0358

Mössbauer study of iron gall inks on historical documents

AL designed and coordinated the practical and scientific work, was involved in the evaluation and interpretation of the data and wrote various drafts of the paper. FEW designed and coordinated the practical and scientific work, performed the Mössbauer spectroscopy, evaluated and interpreted the data and wrote the final version of the manuscript. MD carried out the pH measurements of the papers and determined the nature of the papers and gave valuable hints to research on historical documents. TE provided the fragments of the two Spanish documents. JLPP proposed this investigation and gave valuable hints on the documents. All authors read and approved the final manuscript.Iron gall ink was used in the Western world as a permanent writing material already in late Roman times and throughout the Middle Ages, until it became obsolete in the twentieth century. There is much interest in experimental methods to determine the state of the ink and its degradation products on historical documents. Mossbauer spectroscopy with Fe-57 is such a method, and it has the particular advantage to be sensitive to the chemical bonding of iron, but this method has only rarely been applied to historical documents. In this paper we present Mossbauer data for two damaged documents from a Library in Granada and a handwritten German book from the eighteenth century. In addition to the inked parts of the manuscripts, ink-free parts were studied to determine the amount and chemical state of the iron in the papers. These new results are discussed in the context of previously published Mossbauer data. In one of the investigated documents Fe(II)-oxalate, FeC2O4 center dot 2H(2)O, was observed. The assignment of the various Fe3+ sites in the different documents is rather difficult and often there is a superposition of various species. Known forms of iron gallate are definitely not present on the inked papers. The observed ferric species can be remains of Fe3+ polyphenol complexes of the ink, complexes of Fe3+ with degradation products of the cellulose of the paper or gum arabic, or very small iron oxide or hydroxide nanoparticles

Application of a robust QFT linear control method to the course changing manoeuvring of a ship

This paper describes in detail the design methodology of a robust QFT (Quantitative Feedback Theory) controller for the control of the course changing of a ship. A linear model is used with uncertainty in the parameters. The system is designed to fulfil the specifications of robust stability and robust tracking of a reference system

Synthesis of Catechol Derived Rosamine Dyes and Their Reactivity toward Biogenic Amines

Functional organic dyes play a key role in many fields, namely in biotechnology and medical diagnosis. Herein, we report two novel 2,3- and 3,4-dihydroxyphenyl substituted rosamines (3 and 4, respectively) that were successfully synthesized through a microwave-assisted protocol. The best reaction yields were obtained for rosamine 4, which also showed the most interesting photophysical properties, specially toward biogenic amines (BAs). Several amines including n- and t-butylamine, cadaverine, and putrescine cause spectral changes of 4, in UV–Vis and fluorescence spectra, which are indicative of their potential application as an effective tool to detect amines in acetonitrile solutions. In the gas phase, the probe response is more expressive for spermine and putrescine. Additionally, we found that methanolic solutions of rosamine 4 and n-butylamine undergo a pink to yellow color change over time, which has been attributed to the formation of a new compound. The latter was isolated and identified as 5 (9−aminopyronin), whose solutions exhibit a remarkable increase in fluorescence intensity together with a shift toward more energetic wavelengths. Other 9-aminopyronins 6a, 6b, 7a, and 7b were obtained from methanolic solutions of 4 with putrescine and cadaverine, demonstrating the potential of this new xanthene entity to react with primary amines.Financial support from PT national funds (FCT/MCTES, Fundação para a Ciência e a Tecnologia and Ministério da Ciência, Tecnologia e Ensino Superior) through the project PTDC/QUI-QOR/29426/2017. The research team would like to thank the projects UIDB/50006/2020, PTDC/QUI-QIN/28142/2017 and Grant BU263P18 (from the Junta de Castilla y León, Consejería de Educación y Cultura y Fondo Social Europeo). F. M. -S. gratefully acknowledges FCT (Portugal’s Foundation for Science and Technology) within grant DFA/BD/9136/2020. A.M.G.S. and A.L. thank FCT for the program DL 57/2016 – Norma transitória

Effect of angiotensin II and small GTPase Ras signaling pathway inhibition on early renal changes in a murine model of obstructive nephropathy

This is an open access article distributed under the Creative Commons Attribution License.Tubulointerstitial fibrosis is a major feature of chronic kidney disease. Unilateral ureteral obstruction (UUO) in rodents leads to the development of renal tubulointerstitial fibrosis consistent with histopathological changes observed in advanced chronic kidney disease in humans. The purpose of this study was to assess the effect of inhibiting angiotensin II receptors or Ras activation on early renal fibrotic changes induced by UUO. Animals either received angiotensin II or underwent UUO. UUO animals received either losartan, atorvastatin, and farnesyl transferase inhibitor (FTI) L-744,832, or chaetomellic acid A (ChA). Levels of activated Ras, phospho-ERK1/2, phospho-Akt, fibronectin, and α-smooth muscle actin were subsequently quantified in renal tissue by ELISA, Western blot, and/or immunohistochemistry. Our results demonstrate that administration of angiotensin II induces activation of the small GTPase Ras/Erk/Akt signaling system, suggesting an involvement of angiotensin II in the early obstruction-induced activation of renal Ras. Furthermore, upstream inhibition of Ras signalling by blocking either angiotensin AT1 type receptor or by inhibiting Ras prenylation (atorvastatin, FTI o ChA) reduced the activation of the Ras/Erk/Akt signaling system and decreased the early fibrotic response in the obstructed kidney. This study points out that pharmacological inhibition of Ras activation may hold promise as a future strategy in the prevention of renal fibrosis.This study was supported by grants from Ministerio de Economía y Competitividad (Grant SAF2010-15881 and Red de Investigacion Cooperativa en Enfermedades Renales REDINREN RD12/0021/0032), Junta de Castilla y León (Grant SA 001/C05 and Excellence Group GR100), and REDINREN which is an initiative of the Instituto de Salud Carlos III of Spain supported by FEDER funds. When performing the present study, Ana B. Rodríguez-Pena was a fellow of the Fundacion Renal “Iñigo Ávarez de Toledo” and Neil G. Docherty was a fellow ofThe Marie Curie Programme, EU.Peer Reviewe

Obtención de biomasa de microalga Chlorella vulgaris en un banco de prueba de fotobiorreactores de columna de burbujeo

Dada la agudización de la situación socioeconómica y medioambiental que se enfrenta en la actualidad, los investigadores buscan nuevas alternativas para sustituir el combustible fósil convencional, siendo una salida, los biocombustibles obtenidos a partir de microalgas. El objetivo de esta investigación fue la obtención de biomasa en un banco de prueba de fotobiorreactores de columna de burbujeo, utilizando una cepa de Chlorella vulgaris en medio Bristol. Se dimensionó el fotobiorreactor y se evaluó la influencia de las variables pH y concentración de nitrógeno, con y sin presencia de oligoelementos, sobre la productividaddel crecimiento de la biomasa de microalgas. Seencontró que en el intervalo estudiado (pH entre 6 y 8 y concentración de NaNO3 entre 0,5 y 1 g/L) estas variables no tienen un efecto significativo en el crecimiento, mientras que la presencia de oligoelementos favorece este

Producción de ganado bovino de carne con bienestar animal

Objective: To review several factors related to animal welfare which affect beef cattle productivity and meat quality. Approach: The human population increase has risen the demand for animal products. Therefore it has been necessary to produce them in an intensive way, without considering animal welfare. Nowadays beef cattle producers are aware of the different factors that affect animal welfare, such as nutrition, environmental conditions, housing, handling, diseases, transportation, and slaughter; because national and international markets demand animal products in which animal welfare has been taken into account, are innocuous and have high quality for the final consumerObjetivo: Revisar diferentes factores relacionados con bienestar animal que afectan la producción y calidad de carne en bovinos. Aproximación: El aumento de la población mundial ha incrementado la demanda de productos de origen animal, siendo necesario producirlos en forma intensiva, sin tomar en cuenta el bienestar animal. Actualmente los productores de ganado para carne están cada vez más conscientes de los diferentes factores que afectan el bienestar como son la nutrición, condiciones ambientales, instalaciones, manejo, enfermedades, transporte y sacrificio, ya que los mercados nacionales e internacionales demandan productos obtenidos bajo normas de bienestar animal,garantizando productos cárnicos inocuos y de alta calidad para los consumidores finales

Análisis arqueosismológico del conjunto arqueológico romano de Mulva- Munigua (Sevilla, España). Resultados preliminares

El conjunto arqueológico romano de Mulva-Munigua (Sevilla, España) presenta daños en las edificaciones que pueden ser interpretadas como resultado de la ocurrencia de un evento sísmico (Efectos Arqueológicos de los Terremotos: EAEs) a finales del siglo III A.D., fecha coincidente con el inicio del periodo de declive económico de este asentamiento romano. Para intentar establecer el posible origen sísmico de las deformaciones, se ha procedido al inventario y análisis de las estructuras deformadas presentes en el yacimiento. No obstante, algunas de estas deformaciones también se pueden interpretar como resultado de procesos gravitaciones asociados a la ladera Este de la colina sobre la que se sitúa parte del yacimiento. Las direcciones de máxima deformación (ey) obtenidas del análisis de EAEs indica dos direcciones preferentes de la deformación (o movimiento preferente del terreno): NNO-SSE y ENEOSO. Aunque los datos presentan una dispersión importante, se puede establecer que la orientación principal NNO-SSE es compatible con un evento sísmico situado en el borde norte del Valle del Guadalquivir. La orientación ENE-OSO podría relacionarse con un evento posterior, o más seguramente con procesos de ladera de carácter cosísmico o no.The Roman archaeological site of Mulva-Munigua (Sevilla, Spain) displays building damage features suggesting a seismic origin (Earthquake Archaeological Effects: EAEs). The proposed seismic event could be tentatively dated in the late 3rd century AD, coinciding with the beginning of the economic fall of the Roman Empire at Iberia. However, some of the recorded EAEs can be also interpreted as a result of intervening slope movements in the eastern hillslope of this roman site. The inventory and analysis of the proposed EAEs make possible to discern between seismic oriented damage and other causes. In spite of the data show a significant dispersion, their analysis result in two different orientations of maximum deformation (ey) or preferential ground movement: NNW-SSE and ENE-WSW. The main ey orientation (NNW-SSE) can be tentatively related to a seismic event occurred in the environs of the northern border of the Guadalquivir Depression. The secondary orientation (ENE-WSW) can be interpreted as a consequence of latter slope movements triggered (or not) by other ancient earthquakes