Isolation and identification by high resolution liquid chromatography tandem mass spectrometry of novel peptides with multifunctional lipid-lowering capacity

This work describes the isolation, characterization, and identification by RP-HPLC-ESI-Q-TOF of novel peptides that interfere in the fat digestion and absorption mechanisms by multiple pathways. Peptides were ultrafiltrated and peptides in the most active fraction were further separated by semipreparative RP-HPLC. Nine different subfractions were obtained observing a high amount of peptides in subfraction F3. Peptides in subfraction F3 could simultaneously reduce the solubility of cholesterol in micelles and inhibit pancreatic cholesterol esterase and pancreatic lipase, even after a simulated gastrointestinal digestion. The identification of lipid-lowering peptides has been scarcely performed and when done, low selectivity or sensitivity of employed identification techniques or conditions did not yield reliable results. Separation and detection of peptides by RP-HPLC-ESI-Q-TOF-MS was optimized and most favorable conditions were employed for the identification of peptides using de novo sequencing. Ten different peptides with 4-9 amino acids were identified. Main feature of identified peptides was the high acidity derived from a high presence of amino acids glutamic acid and aspartic acid in their sequences

Preparation of inocula for experimental infection of blood with Streptococcus pneumoniae

Las infecciones experimentales de cualquiera de las células requieren la preparación de buena calidad inóculos. Desafortunadamente, el patógeno pulmonar Streptococcus pneumoniae es un microorganismo que sufre un proceso de autolisis cuando se cultiva in vitro. La suplementación de caldo de Todd-Hewitt con un tampón biológico (20 mM Tris-HCl, pH = 7.8) promueve un retraso de seis horas en el comienzo del proceso de autólisis. Hay mejoras cuando hay lavado de bacterias antes de la congelación, evitando manipulaciones después dedescongelación, y el uso de glicerol ( 70% de la bacterias congeladas era viable después de 28 semanas a -80 ° C, y las alícuotas fueron altamente homogéneo. Hemos probado su utilidad en un modelo de infección de la sangre entera y tienen encontró que el plasma humano exhibe una actividad microbicida superior a la sangre entera, una resultado de que no hemos encontrado informado anteriorment

Identification of Peptides Potentially Responsible for In Vivo Hypolipidemic Activity of a Hydrolysate from Olive Seeds

Previous studies demonstrated that peptides produced by the hydrolysis of olive seed proteins using Alcalase enzyme showed in vitro multifunctional lipid-lowering capability. This work presents a deeper insight into the hypolipidemic effect of olive seed peptides. The capability of olive seed peptides to inhibit endogenous cholesterol biosynthesis through the inhibition of HMGCoA reductase enzyme was evaluated observing a 38 ± 7% of inhibition. Two in vivo assays using different peptides concentrations (200 and 400 mg/kg/day) were designed to evaluate the hypolipidemic effect of olive seed peptides in male and female mice. A low concentration of hydrolysate reduced total cholesterol in male mice in a 20% after 11 weeks compared to the mice feeding with hypercholesterolemic diet. A higher hydrolysate concentration showed a greater reduction in total cholesterol (25%). The analysis of the olive seed hydrolysate by reverse phase high-performance liquid chromatography mass spectrometry (RP-HPLC-MS) enabled the identification of peptides that could be responsible for this hypolipidemic effect

Evaluation of the relationship between the peptide profile and the lipid-lowering properties of olive seeds hydrolysates as a tool for tunning hypocholesterolemic functionality

Olive processing generates large amounts of stones with high protein contents. Previous studies have demonstrated that Manzanilla variety olive seed proteins release peptides with lipid-lowering capacity. However, no work has demonstrated their roles in the overall hypolipidemic activity. Moreover, further studies using different olive varieties are required to propose a solid method for the exploitation of olive seeds. Twenty different olive varieties were employed in this work. Proteins were extracted using high-intensity focused ultrasound and digested with Alcalase. The released peptides were identified using proteomic techniques, and their capabilities to reduce the absorption of dietary cholesterol (by inhibiting cholesterol esterase enzyme, binding bile acids, and reducing micellar cholesterol solubility) or the biosynthesis of endogenous cholesterol were evaluated. Peptides with different lipid lowering capacities were obtained from all varieties although the genotype significantly affected the hypolipidemic characteristics. Univariate and multivariate statistical analyses showed strong correlations, positive and negative, between the presence of certain peptides in the hydrolysates and their capacity to reduce exogenous cholesterol absorption and endogenous cholesterol synthesis. Therefore, the selection of the olive seed genotype can direct its lipid-lowering properties,e.g., by promoting the reduction of dietary cholesterol absorption or the inhibition of cholesterol biosynthesis

Magnetic nanoparticles coated with carboxylate-terminated carbosilane dendrons as a reusable and green approach to extract/purify proteins

Extraction/purification of proteins, at both analytical and industrial levels, is a limiting step that usually requires the use of organic solvents and involves tedious work and a high cost. This work proposes a more sustainable alternative based on the use of magnetic nanoparticles (MNPs) coated with carboxylate-terminated carbosilane dendrons. MNPs coated with first- and second-generation carbosilane dendrons and bare MNPs were employed for the extraction of proteins with different molecular weights and charges. Interaction of proteins with MNPs significantly varied with the pH, the protein, and the dendron generation (different sizes and number of charges in the periphery). Optimal dendron:protein molar ratios and suitable conditions for disrupting interactions after protein extraction were also researched. Second-generation dendron-coated MNPs showed 100% retention capability for all proteins when using acidic conditions. They were reused without losing magnetism or interaction capacity after a disruption of protein-dendron interactions with 0.2% SDS at 100 degrees C for 10 min. The capacity of dendron-coated MNPs was successfully applied to the recovery/purification of proteins from two food byproducts, olive seeds and cheese whey

The role of snow in scavenging aerosol particles: A physical-chemical characterization

[EN] The below cloud scavenging of aerosols by snow has been analysed in León (NW Spain). Six snow events were registered over the course of one year of study. Ultrafine and accumulation aerosol particles were measured using a scanning mobility particle sizer spectrometer, while hydrometeors were characterized using a disdrometer. Furthermore, the chemical composition of the melted snow-water samples (soluble and insoluble fractions) was analysed. The scavenging coefficient (λ) showed a great variability among events. An effective washing of particles was observed during the first 30 min of snowfall. The mean change in the scavenging efficiency (%ΔC) of particle number concentration (PNC) and λ coefficient during this time interval were: i) nucleation mode: 36.3 % and 3.02 · 10−4 s−1; ii) Aitken mode: 30.4 % and 2.37 · 10−4 s−1 and iii) accumulation mode: 22.4 % and 1.77 · 10−4 s−1. The range of particle sizes that is less efficiently scavenged by snowfall was observed between 400 and 600 nm. When analyzing the whole snow event, an increase of PNC was observed. Two possible explanations underlie this behaviour: it could be caused by changes in air masses or by the resuspension of aerosol particles scavenged by snowflakes upon reaching the ground. A clear relationship was observed between Ca2+, SO42− and NO3− concentrations of aerosol particles before the snow event and the concentrations registered in the melted snow-water. The largest and smallest changes in aerosol number concentrations were caused by snowflakes of 3 and 6 mm in diameter, respectively. The particle size distributions (PSD) were fitted to log-normal distributions and the parameters were compared before and after snowfall.S

BRAF V600E mutational load as a prognosis biomarker in malignant melanoma

Analyzing the mutational load of driver mutations in melanoma could provide valuable information regarding its progression. We aimed at analyzing the heterogeneity of mutational load of BRAF V600E in biopsies of melanoma patients of different stages, and investigating its potential as a prognosis factor. Mutational load of BRAF V600E was analyzed by digital PCR in 78 biopsies of melanoma patients of different stages and 10 nevi. The BRAF V600E load was compared among biopsies of different stages. Results showed a great variability in the load of V600E (0%-81%). Interestingly, we observed a significant difference in the load of V600E between the early and late melanoma stages, in the sense of an inverse correlation between BRAF V600E mutational load and melanoma progression. In addition, a machine learning approach showed that the mutational load of BRAF V600E could be a good predictor of metastasis in stage II patients. Our results suggest that BRAF V600E is a promising biomarker of prognosis in stage II patients.This research was supported by the Basque Government (grants ELKARTEK-KK2016-036 and KK2017-041 to MDB, grant IT1138-16 to SA and predoctoral fellowship PRE_2014_1_419 to AS), and by the University of the Basque Country (UPV/EHU) (grant GIU17/066). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript

Art and press, innovation in the language of communication

[EN] “Diario de las Bellas Artes”. Diario de Teruel. It is a collaboration between Diario de Teruel and the Grado en Bellas Artes of Universidad de Zaragoza. In a pioneer manner –for the second consecutive year– photographs of all the news of the day are replaced by illustrations made by students and teachers of the degree, incorporating enhanced reality, animations and interactivity inside the pages. “Diario de las Bellas Artes” seeks innovation in traditional newspapers, through new technologies and multimedia. In agroundbreaking way, it incorporates interactive, multimedia and augmented reality news in written press. You can get access to the interactivity of the paper issue by using internet from your smartphone, tablet or computer. The two previous editions of “Diario de las Bellas Artes”. Diario de Teruel are available in the following link: http://diariodelasbellasartes.blogspot.es[ES] “Diario de las Bellas Artes”. Diario de Teruel. It is a collaboration between Diario de Teruel and the Grado en Bellas Artes of Universidad de Zaragoza. In a pioneer manner –for the second consecutive year– photographs of all the news of the day are replaced by illustrations made by students and teachers of the degree, incorporating enhanced reality, animations and interactivity inside the pages. “Diario de las Bellas Artes” seeks innovation in traditional newspapers, through new technologies and multimedia. In agroundbreaking way, it incorporates interactive, multimedia and augmented reality news in written press. You can get access to the interactivity of the paper issue by using internet from your smartphone, tablet or computer. The two previous editions of “Diario de las Bellas Artes”. Diario de Teruel are available in the following link: http://diariodelasbellasartes.blogspot.esHernández Muñoz, S.; López Alonso, F.; López Juderías, C.; Muñoz Tomás, I.; Prieto, J.; Hernando Sebastián, PL.; Benedicto, R.... (2017). Arte y prensa, innovando en el lenguaje de la comunicación. En Glocal [codificar, mediar, transformar, vivir] III Congreso Internacional de Investigación en Artes Visuales. Editorial Universitat Politècnica de València. 339-347. https://doi.org/10.4995/ANIAV.2017.5712OCS33934

Depth of response in multiple myeloma: A pooled analysis of three PETHEMA/GEM clinical trials

On behalf of the GEM (Grupo Español de Mieloma)/PETHEMA (Programa para el Estudio de la Terapéutica en Hemopatías Malignas) Cooperative Study Group.[Purpose]: To perform a critical analysis on the impact of depth of response in newly diagnosed multiple myeloma (MM). [Patients and Methods]: Data were analyzed from 609 patients who were enrolled in the GEM (Grupo Español de Mieloma) 2000 and GEM2005MENOS65 studies for transplant-eligible MM and the GEM2010MAS65 clinical trial for elderly patients with MM who had minimal residual disease (MRD) assessments 9 months after study enrollment. Median follow-up of the series was 71 months. [Results]: Achievement of complete remission (CR) in the absence of MRD negativity was not associated with prolonged progression-free survival (PFS) and overall survival (OS) compared with near-CR or partial response (median PFS, 27, 27, and 29 months, respectively; median OS, 59, 64, and 65 months, respectively). MRD-negative status was strongly associated with prolonged PFS (median, 63 months; P operational cure> was high; median PFS was 12 years, and the 10-year OS rate was 94%. [Conclusion]: Our results demonstrate that MRD-negative status surpasses the prognostic value of CR achievement for PFS and OS across the disease spectrum, regardless of the type of treatment or patient risk group. MRD negativity should be considered as one of the most relevant end points for transplant-eligible and elderly fit patients with MM.Supported by the Centro de Investigación Biomédica en Red – Area de Oncologia - del Instituto de Salud Carlos III (CIBERONC; CB16/12/00369; CB16/12/00400; CB16/12/00233; CB16/12/00284), formerly named as Cooperative Research Thematic Network (Grants No. RD12/0036/0058, RD12/0036/0048, RD12/0036/0046, and RD12/0036/0061) of the Red de Cancer (Cancer Network of Excellence); Instituto de Salud Carlos III/Subdirección General de Investigación Sanitaria; funded in part by the European Regional Development Fund (FIS No. 98/1239, 00/10160, 01/0089, 02/0089, 02/0905, G03/136, PI051284, PI06033906/1354, PS09/01897/01370, PI12/01761, PI12/02311, PI13/01469, PI14/01867, G03/136); Sara Borrell (No. CD13/00340); Asociación Española Contra el Cáncer (No. GCB120981SAN); and Federación Española de Enfermedades Raras. Also supported internationally by the Black Swan Research Initiative of the International Myeloma Foundation and the European Research Council 2015 Starting Grant (MYELOMANEXT).Peer Reviewe