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

Porous Titanium Cylinders Obtained by the Freeze-Casting Technique: Influence of Process Parameters on Porosity and Mechanical Behavior

 The discrepancy between the stiffness of commercially pure titanium and cortical bone  tissue compromises its success as a biomaterial. The use of porous titanium has been widely studied,  however, it is still challenging to obtain materials able to replicate the porous structure of the bones  (content, size, morphology and distribution). In this work, the freeze‐casting technique is used to  manufacture cylinders with elongated porosity, using a home‐made and economical device. The  relationship between the processing parameters (diameter and material of the mold, temperature  gradient), microstructural features and mechanical properties is established and discussed, in terms  of ensuring biomechanical and biofunctional balance. The cylinders have a gradient porosity  suitable for use in dentistry, presenting higher Young’s modulus at the bottom, near the cold spot  and, therefore better mechanical resistance (it would be in contact with a prosthetic crown), while  the opposite side, the hot spot, has bigger, elongated pores and walls.  Ministry of Economy and Competitiveness of Spain  grant  MAT2015‐71284‐P  FEDER‐Junta de Andalucía Research  Project (Modeling and implementation of the freeze casting technique: gradients of porosity with a tribomechanical equilibrium and electro‐stimulated cellular behavior).

Increased plasma soluble endoglin levels as an indicator of cardiovascular alterations in hypertensive and diabetic patients

<p>Abstract</p> <p>Background</p> <p>Endoglin is involved in the regulation of endothelial function, but there are no studies concerning its relation with hypertension- and diabetes-associated pathologies. Thus, we studied the relationship between plasma levels of soluble endoglin and cardiovascular alterations associated with hypertension and diabetes.</p> <p>Methods</p> <p>We analyzed 288 patients: 64 with type 2 diabetes, 159 with hypertension and 65 healthy patients. We assessed the relationship of soluble endoglin plasma levels measured by enzyme-linked immunosorbent assay with basal glycemia, glycosylated hemoglobin, blood pressure, endothelial dysfunction (assessed by pressure wave velocity), hypertensive retinopathy (by Keith-Wagener classification), left ventricular hypertrophy (by Cornell and Sokolow indexes), cardiovascular risk and target organ (heart, vascular, kidney) damage.</p> <p>Results</p> <p>There are significant correlations between endoglin and glycemia, systolic blood pressure, pulse pressure, pressure wave velocity and electrocardiographically assessed left ventricular hypertrophy. Endoglin levels were significantly higher in patients with diabetes who had nondipper and extreme dipper circadian blood pressure patterns than in dipper circadian patterns, in patients with hypertension and diabetes who had riser pattern than in the other patients, and in patients with diabetes but not hypertension who had extreme dipper pattern than in dipper, nondipper and riser groups. There was also a significant correlation between plasma-soluble endoglin and lower levels of systolic night-day ratio. Higher endoglin levels were found in patients with diabetes who had retinopathy, in patients with diabetes who had a high probability of 10-year cardiovascular risk, and in patients with diabetes and hypertension who had three or more damaged target organs (heart, vessels, kidney) than in those with no organs affected.</p> <p>Conclusions</p> <p>This study shows that endoglin is an indicator of hypertension- and diabetes-associated vascular pathologies as endothelial dysfunction and cardiovascular damage.</p

Balancing porosity and mechanical properties of titanium samples to favor cellular growth against bacteria

Two main problems limit the success of titanium implants: bacterial infection, which restricts their osseointegration capacity; and the stiffness mismatch between the implant and the host cortical bone, which promotes bone resorption and risk of fracture. Porosity incorporation may reduce this difference in stiffness but compromise biomechanical behavior. In this work, the relationship between the microstructure (content, size, and shape of pores) and the antibacterial and cellular behavior of samples fabricated by the space-holder technique (50 vol % NH4HCO3 and three ranges of particle sizes) is established. Results are discussed in terms of the best biomechanical properties and biofunctional activity balance (cell biocompatibility and antibacterial behavior). All substrates achieved suitable cell biocompatibility of premioblast and osteoblast in adhesion and proliferation processes. It is worth to highlighting that samples fabricated with the 100–200 μm space-holder present better mechanical behavior—in terms of stiffness, microhardness, and yield strength—which make them a very suitable material to replace cortical bone tissues. Those results exposed the relationship between the surface properties and the race of bacteria and mammalian cells for the surface with the aim to promote cellular growth over bacteria.University of Seville (Spain) VI Plan Propio de Investigación y Transferencia—US 2018, I.3A

Progradation of a shallow carbonate platform developed on a fault-block in the Western Tethys (lower Aptian, Sierra de Bedmar-Jódar, Prebetic of Jaén, Spain).

The Middle Member of the Llopis Fm in the Sierra de Bedmar-Jódar Unit of the Prebetic Zone of Jaén (southern Spain) was deposited on a shallow-marine platform of the Southern Iberian Continental Margin during the earliest Aptian. Detailed field logging of nine stratigraphic sections and facies mapping have allowed seven lithofacies associations (L1–L6) to be distinguished, one siliciclastic (L1) and five carbonate facies (L2–6). The succession is composed of eight consecutive elemental sequences of lithofacies associations L1–L6. Each elemental sequence is interpreted as representing one episode of shallowing-upwards carbonate deposition in a very shallow platform-lagoon that was bounded shoreward by clastic/ooid bars and passed seaward either to stromatoporoid bioconstructions (bioherms and biostromes) or rudist biostromes. The successive elemental sequences show north-eastward progradational geometries. Three phases of platform development are identified: (1) installation of the shallow platform; (2) development of a lagoon bounded by a stromatoporoid barrier and (3) development of an Urgonian-type platform dominated by rudists. During the early Aptian, the Bedmar-Jódar platform was partially isolated from the rest of the Prebetic platform and showed overall progradation towards the NE, in contrast to the general south-eastward progradational trend of the Prebetic platform. Sedimentation was controlled by rift-generated extensional tectonics that resulted in tilting of the platform block, causing the deviation of progradation from the general trends of the Prebetic Platform. In addition, climatic influence is inferred from the presence of siliciclastic sediments derived from weathering of the hinterland, which restricted the carbonate factory.Depto. de Geodinámica, Estratigrafía y PaleontologíaFac. de Ciencias GeológicasTRUEUniversidad de JaénSecretaría de Estado de I + D + iJunta de Andalucíapu

Porous titanium substrates coated with a bilayer of bioactive glasses

Porous titanium substrates coated by dripping-sedimentation technique with a novel bilayer of (45S5 / 1393) bioactive glasses are proposed to overcome some limitations of the use of titanium for implants, such as the stress shielding and the poor osseointegration. Composition, thickness, roughness and micromechanical behavior (P-h curves) of the coating and the influence of the porous titanium substrates have been characterized. Best results were found for the substrate with 30 vol.% of porosity and a range size of 355 ‒ 500 μm, since it enhanced the mechanical and biofunctional behavior, due to the good adhesion of the 1393 bioglass to the substrate and the greater bioactivity of the 45S5 bioglass, which would be in contact with the bone.Junta de Andalucía–FEDER (Spain) US-1259771M.E.C. (Spain) 2004/00001203 (RYC-2004-001497

Biofunctional and Tribo-mechanical Behavior of Porous Titanium Substrates Coated with a Bioactive Glass Bilayer (45S5 ‒ 1393)

Porous substrates of commercially pure titanium have been coated with a novel bilayer of bioactive glasses, 45S5 and 1393, to improve the osseointegration and solve the stress-shielding phenomenon of titanium partial implants. The porosity of the substrates, the scratch resistance and bioactivity of the coating have been evaluated. Results are discussed in terms of stiffness and yield strength of the substrates, as well as the chemical composition, thickness and design of the bioglass coating (monolithic vs. bilayer). The role of the pores was a crucial issue in the anchoring of the coating, both in porosity percentage (30 and 60 vol. %) and pore range size (100 – 200 and 355 ‒ 500 μm). The study was focused on the adhesion and infiltration of a 1393 bioglass layer (in contact with a porous titanium substrate), in combination with the biofunctionality of the 45S5 bioglass layer (surrounded by the host bone tissue), as 1393 bioglass enhances the adherence, while 45S5 bioglass promotes higher bioactivity. This bioactivity of the raw powder was initially estimated by nuclear magnetic resonance, through the evaluation of the chemical environments, and confirmed by the formation of hydroxyapatite, when immersed in simulated body fluid. Results revealed that the substrate with 30 vol. % of porosity and a range of 355 ‒ 500 μm pore size, coated with this novel bioactive glass bilayer, presented the best combination in terms of mechanical and biofunctional properties.Junta de Andalucía–FEDER (Spain) Project Ref. US-1259771Ministerio de Ciencia y Educación (Spain) Project 2004/00001203 (RYC-2004-001497

Fabrication and characterization of superficially modified porous dental implants

Stress-shielding and loosening compromise the success of dental implants under real-life service conditions. This work evaluates the mechanical behavior of superficially modified porous titanium dental implants fabricated by two different routes: conventional powder metallurgy and space-holder techniques. A novel, feasible and repetitive protocol of micro-milling of the implant thread (before sintering), as well as surface modification treatments (after sintering) are also implemented. The discussion is conducted in terms of the influence of porosity and surface roughness on the stiffness and yield strength of implants. The macro-pores concentrate stress locally, and, at the same time, they could act as a barrier to the propagation of micro-cracks. Higher rugosity was observed for virgin implants obtained with spacer particles. Concerning superficially modified implants, while bioglass 1393 was the most effective coating due to its greater infiltration and adhesion capacity, chemical etching could improve osteoblast adhesion because modifies the roughness of the implant surface.Ministry of Science and Innovation of Spain PID2019-109371GB-I00Junta de Andalucía–FEDER (Spain) US-125977

Tracking magmatism and oceanic change through the early Aptian Anoxic Event (OAE 1a) to the late Aptian: Insights from osmium isotopes from the westernmost Tethys (SE Spain) Cau Core

Some of the major Carbon cycle perturbations of the Phanerozoic occurred during the Aptian, in relation to magmatism. The highest temperatures reconstructed for the Cretaceous Period correspond to the Oceanic Anoxic Event of the early Aptian (OAE 1a), an episode of accelerated global change. Here we present a chemostratigraphic study based on osmium isotopes integrated with high-resolution Carbon-Oxygen stable isotope data from the Cau Core (Western Tethys, SE Spain), including a 6.4 Ma record from the early to the late Aptian. This high-resolution study of the continuous and expanded Cau section permits a thorough understanding of the duration of the Aptian events, as well as an evaluation of the mechanisms triggering the abrupt changes of the global carbon and osmium cycles and their interdependence. Here we show that the Large Igneous Province (LIP) Aptian magmatism initiated 550–750 kyr prior to the OAE 1a, and persisted for 1.4 Myr after the event, influencing the composition of seawater for 2.8 Myr. We show a continuous Os isotope record encompassing the OAE 1a and the late Aptian for the first time, and demonstrate that the recovery from the exceptionally unradiogenic composition of seawater Os produced by the dominance of the Ontong Java Plateau volcanism, was slow. Our results demonstrate the different time duration of some events, and the asynchronous relationship between the carbon and osmium cycles