Main Variables Affecting a Chemical-Enzymatic Method to Obtain Protein and Amino Acids from Resistant Microalgae

he development of microalgae uses requires further investigation in cell disruption alternatives to reduce the costs associated to this processing stage. This study aimed to evaluate the main variables affecting an extraction method to obtain protein and amino acids from microalgae. The method was based on a sequential alkaline-enzymatic process, with separate extractions and noncontrolled pH, and was applied to fresh biomass of a resistant species. The processed microalgae were composed of a consortium with Nannochloropsis sp. as predominant species. After the optimization of the pH of the alkaline reaction, the effect of the time of the alkaline reaction (30-120min), the time (30-120min) and temperature (40-60 degrees C) of the enzymatic reaction, and the biomass concentration (50-150mgml(-1)), on the extraction yields of protein and free amino nitrogen (FAN) and on the final concentration of protein in the extract, was studied using a response surface methodology. Even though all the variables and some interactions among them had a significant effect, the biomass concentration was the most important factor affecting the overall process. The results showed relevant information about the different options in order to maximize not only the response variables individually but also different combinations of them. Assays with optimized values reached maximum yields of 80.3% and 1.07% of protein (% of total protein) and FAN (% of total biomass), respectively, and a protein concentration in the extract of 15.2mgml(-1). The study provided the essential information of an alternative approach to obtain protein and amino acids from fresh biomass of resistant microalgae with a high yield, also opening perspectives for further research in particular aspects

Estudio baropodométrico en pacientes tratados mediante artroplastia total de rodilla

La patología artrósica de la rodilla condiciona una disminución de la carga del miembro afecto y un patrón de apoyo con predominio generalmente del arco externo. El propósito de este estudio es determinar si existe una modificación significativa de este patrón tras la implantación de una artroplastia total de rodilla (ATR) primaria, recogiendo las presiones plantares de ambos pies mediante baropodometría electrónica previamente y 4 meses tras la cirugía, tanto en estática como en dinámica. Así, se analizan las presiones plantares de 30 pacientes, 21 mujeres y 9 hombres, intervenidos de ATR primaria en el Hospital Universitario Doctor Peset entre los años 2013 y 2015. También se ha determinado si existe una corrección del eje anatómico tras la cirugía. Utilizando test no paramétricos (test de Wilcoxon para datos apareados, U de Mann-Whitney), observamos una modificación significativa de dicho ángulo desde valores medianos de 1.2º de varo a 4.9º de valgo en el postoperatorio, una disminución mediana de la presión plantar en ambos pies y una redistribución de la carga axial, aumentando en el miembro intervenido. Concluimos que, 4 meses tras la cirugía, se produce una tendencia a la “normalización” de la pisada en pacientes en los que se ha implantado una ATR, condicionado tanto por la mejora del dolor y la funcionalidad como por la realineación del eje.Knee osteoarthritis determines a decrease of the load of the affected limb and a pattern usually dominance of external support plantar arc. The purpose of this study is to determine whether a significant change in this pattern is? after implantation of a primary total knee arthroplasty (TKA), measuring plantar pressures in both feet by electronic baropodometry previously and 4 months after surgery, in static and dynamic. Plantar pressures of 30 patients are analyzed, who underwent primary TKA at the University Hospital Doctor Peset between 2013 and 2015. In addition, it has also been determined if there is a correction on the anatomical axis after surgery. Using nonparametric test (Wilcoxon test for paired data, U Mann Whitney), we observed a significant change in the angle from median values of 1.2º varus to 4.9º valgus postoperatively, a median decrease of plantar pressure in both feet and a redistribution of axial load, increasing in the operated limb. We conclude that, 4 months after surgery, there is a tendency to “normalization” of the tread in patients undergoing TKA. This is conditioned by improving pain and function as axis realignment occurs

Application of a Microalgal Peptide-Enriched Extract as Media Component in E. coli Culture

This study evaluated the feasibility of applying a microalgal peptide-enriched extract, obtained through a sequential alkaline and enzymatic method, as a fundamental media component in Escherichia coli culture. The results demonstrated that the extract is viable as the primary nutrient source for E. coli cultures (wild type), even as a complete substitute for the widely used Luria-Bertani (LB) medium, using an extract dilution of 1:3. Its application in a proportion carbon limiting (dilution 1:7), with glycerol supplementation at the early stationary phase, improved the nutrient use efficiency and controlled the undesirable acetate production. Furthermore, this last strategy applied in an E. coli mutant (M4-Delta iclR) enhanced the production of succinate by 67.3% compared with the M9 medium. The innovative approach proposed in this study opens a wide range of applications and studies related to the production of bio-compounds through diverse microorganisms

Greedy Randomized Adaptive Search and Variable Neighbourhood Search for the minimum labelling spanning tree problem

This paper studies heuristics for the minimum labelling spanning tree (MLST) problem. The purpose is to find a spanning tree using edges that are as similar as possible. Given an undirected labelled connected graph, the minimum labelling spanning tree problem seeks a spanning tree whose edges have the smallest number of distinct labels. This problem has been shown to be NP-hard. A Greedy Randomized Adaptive Search Procedure (GRASP) and a Variable Neighbourhood Search (VNS) are proposed in this paper. They are compared with other algorithms recommended in the literature: the Modified Genetic Algorithm and the Pilot Method. Nonparametric statistical tests show that the heuristics based on GRASP and VNS outperform the other algorithms tested. Furthermore, a comparison with the results provided by an exact approach shows that we may quickly obtain optimal or near-optimal solutions with the proposed heuristics

Chronostratigraphy and new vertebrate sites from the upper Maastrichtian of Huesca (Spain), and their relation with the K/Pg boundary

The transitional-continental facies of the Tremp Formation within the South-Pyrenean Central Unit (Spain) contain one of the best continental vertebrate records of the Upper Cretaceous in Europe. This Pyrenean area is therefore an exceptional place to study the extinction of continental vertebrates across the Cretaceous/Paleogene (K/Pg) boundary, being one of the few places in Europe that has a relatively continuous record ranging from the upper Campanian to lower Eocene. The Serraduy area, located on the northwest flank of the Tremp syncline, has seen the discovery of abundant vertebrate remains in recent years, highlights being the presence of hadrosaurid dinosaurs and eusuchian crocodylomorphs. Nevertheless, although these deposits have been provisionally assigned a Maastrichtian age, they have not previously been dated with absolute or relative methods. This paper presents a detailed stratigraphic, magnetostratigraphic and biostratigraphic study for the first time in this area, making it possible to assign most vertebrate sites from the Serraduy area a late Maastrichtian age, specifically within polarity chron C29r. These results confirm that the vertebrate sites from Serraduy are among the most modern of the Upper Cretaceous in Europe, being very close to the K/Pg boundary.Spanish Ministry of Economy and Competitiveness (grant numbers CGL2014-53548-P, CGL2015-64422-P and CGL2017-85038-P), cofinanced by the European Regional Development Fund; and by the Department of Education and Science of the Aragonese Government (grant numbers DGA groups H54 and E05), cofinanced by the European Social Fund (ESF). The paleomagnetic study was possible thanks to the complementary grants (beneficiaries of FPU, grant number CGL2010-16447/BTE: Brief Stays and Temporary Transfers, year 2015) supported by the Spanish Ministry of Culture, Education and Sports; and the Laboratory of paleomagnetism of the University of Burgos (Spain). Eduardo Puértolas Pascual is the recipient of a postdoctoral grant (SFRH/BPD/116759/2016) funded by the Fundação para a Ciência e Tecnologia (FCT-MCTES)

Oxidative stress and non-alcoholic fatty liver disease: effects of omega-3 fatty acid supplementation

Aging is a complex phenomenon characterized by the progressive loss of tissue and organ function. The oxidative-stress theory of aging postulates that age-associated functional losses are due to the accumulation of ROS-induced damage. Liver function impairment and non-alcoholic fatty liver disease (NAFLD) are common among the elderly. NAFLD can progress to non-alcoholic steatohepatitis (NASH) and evolve to hepatic cirrhosis or hepatic carcinoma. Oxidative stress, lipotoxicity, and inflammation play a key role in the progression of NAFLD. A growing body of evidence supports the therapeutic potential of omega-3 polyunsaturated fatty acids (n-3 PUFA), mainly docosahaexenoic (DHA) and eicosapentaenoic acid (EPA), on metabolic diseases based on their antioxidant and anti-inflammatory properties. Here, we performed a systematic review of clinical trials analyzing the efficacy of n-3 PUFA on both systemic oxidative stress and on NAFLD/NASH features in adults. As a matter of fact, it remains controversial whether n-3 PUFA are effective to counteract oxidative stress. On the other hand, data suggest that n-3 PUFA supplementation may be effective in the early stages of NAFLD, but not in patients with more severe NAFLD or NASH. Future perspectives and relevant aspects that should be considered when planning new randomized controlled trials are also discussed

Heterozygous rare genetic variants in non-syndromic early-onset obesity

BACKGROUND: Obesity is a very heterogeneous disorder at both the clinical and molecular levels and with high heritability. Several monogenic forms and genes with strong effects have been identified for non-syndromic severe obesity. Novel therapeutic interventions are in development for some genetic forms, emphasizing the importance of determining genetic contributions. OBJECTIVE: We aimed to define the contribution of rare single-nucleotide genetic variants (RSVs) in candidate genes to non-syndromic severe early-onset obesity (EOO; body mass index (BMI) >+3 standard deviation score, <3 years). METHODS: Using a pooled DNA-sequencing approach, we screened for RSVs in 15 obesity candidate genes in a series of 463 EOO patients and 480 controls. We also analysed exome data from 293 EOO patients from the "Viva la Familia" (VLF) study as a replication dataset. RESULTS:Likely or known pathogenic RSVs were identified in 23 patients (5.0%), with 7 of the 15 genes (BDNF, FTO, MC3R, MC4R, NEGR1, PPARG and SIM1) harbouring RSVs only in cases (3.67%) and none in controls. All were heterozygous changes, either de novo (one in BDNF) or inherited from obese parents (seven maternal, three paternal), and no individual carried more than one variant. Results were replicated in the VLF study, where 4.10% of probands carried RSVs in the overrepresented genes. RSVs in five genes were either absent (LEP) or more common in controls than in cases (ADRB3, LEPR, PCSK1 and PCSK2) in both obese datasets. CONCLUSIONS: Heterozygous RSVs in several candidate genes of the melanocortin pathway are found in ~5.0% patients with EOO. These results support the clinical utility of genetic testing to identify patients who might benefit from targeted therapeutic intervention.Clara Serra-Juhé, Gabriel Á. Martos-Moreno, Francesc Bou de Pieri, Raquel Flores, Julie A. Chowen, Luis A. Pérez-Jurado, Jesús Argent