Tri-Reagent Homogenate Is a Suitable Starting Material for UHPLC-MS Lipidomic Analysis

Background: Transcriptomic and lipidomic dual analyses usually initiate with independent extractive procedures. That entails a difficulty in aligning results from both omics platforms, especially in the case of highly heterogeneous tissues, such as the kidney. Methods: Bligh and Dyer lipid extraction was performed using rat kidney homogenates prepared in PBS or commercially available Tri-reagent used for RNA extraction. Samples were analyzed by ultrahigh performance liquid chromatography-mass spectrometry (UHPLC-MS) lipidomic analysis. Results: Comparison of the lipidome obtained from phosphate-buffered saline (PBS) and Tri-reagent homogenates showed qualitative and quantitative validity of the Tri-reagent homogenate with the exception of ether lipids; the acidic nature of the mix seems to promote the hydrolysis of the ether bond, especially in plasmalogens. We tested several conditions in the sample processing, which allowed to optimize the procedure. Conclusions: Aiming to implement a method that allows the extraction of RNA and lipids from the same tissue homogenate not using external tracers, we here report the use of Tri-reagent homogenates as a suitable starting material for UHPLC-MS lipidomic analysis.This research was funded by the Basque Government (grants IT971-16, IT1476-22 and KK-2020/00069) and the Spanish Ministry of Science and Innovation (PID2021-124425OB-100)

Otolith microchemistry reveals contrasting intra-stock structuring in two demersal species off the Iberian Peninsula

Trabajo presentado en el 7th International Otolith Symposium, celebrado en Viña del Mar (Chile), entre el 9 y el 13 de octubre de 2023.Peer reviewe

Comparative study of the lipid profile of tears and plasma enriched in growth factors.

[EN] The Tear Film Lipid Layer (TFLL) acts primarily as an interface between the aqueous layer and air. Tear film lipid is composed of a thin layer of polar lipids that interact with the secretory layer of the underlying mucosa and a thicker layer of non-polar lipids at the air interface. The tear film has a complex structure and composition that protects the cornea, promotes wound healing, and maintains high-quality vision. Plasma Rich in Growth Factor (PRGF) eye drops emerged as an exciting new treatment for corneal epitheliopathies, including aqueous deficient dry eye. The purpose of this study was to compare the lipidomic profile of eye drops obtained from PRGF with tear lipidome to determine whether PRGF drops could be an adequate complement to tears in patients with impaired TFLL. To address this study, tears and blood was collected and processed from healthy donors to obtain PRGF eye drops. Samples were aliquoted and stored at -80°C until use. The lipid profiles of these samples were analysed by Ultrahigh Performance Liquid Chromatography (UHPLC) using a Vanquish UHPLC system to obtain untargeted lipidome profiles on a Q-Exactive HF-X hybrid quadrupole-Orbitrap mass spectrometer. In PRGF eye drops, 408 lipids were identified in ESI+mode and 183 in ESI- mode, and they were grouped into 15 different lipid classes from four distinct categories. By contrast, 112 lipid species were identified from tear samples in ESI+mode and 36 in ESI- mode, belonging to 12 lipid classes from six different categories. The relative abundance of most lipid species was much greater in the PRGF eye drops than in the tear, although there were some lipids present in tears that were not found in the PRGF, such as wax esters and (O-acyl)-omega-hydroxy fatty acids. In summary, these results suggest that the lipids present in PRGF eye drops could serve as a tear supplement in individuals in whom tear lipid composition is altered, although there are differences in the lipid profile of these two fluids.The authors wish to acknowledge the financial support received to carry out this work from: MINECO-Retos Fondos Fender (RTC-2016-48231), Gobierno Vasco (PUE_2018_1_0004), ELKARTEK (KK-2019/00086), PIBA 2020-1-0026, MINECO-Retos (PID2019-111139RB-I00), ELKARTEK (KK-2021-00023) to EV and FISS-21-RD21/0002/0041 to AA

Roma en Red: docencia transversal entre 3 disciplinas y aprendizaje colaborativo para la creación y difusión on-line de contenidos didácticos comunes

Depto. de Derecho Romano e Historia del DerechoFac. de DerechoFALSEsubmitte

Measurement invariance of the day reconstruction method: Results from the COURAGE in Europe Project

The final publication is available at Springer via http://dx.doi.org/10.1007/s10902-015-9669-xGiven the growing interest in the study of subjective well-being as a measure of social progress, instruments that produce valid and reliable scores and that can be used within and across countries are needed. The aim of the present study was to analyze the measurement equivalence of the Day Reconstruction Method in its brief version, using nationally representative samples from Finland, Poland, and Spain obtained within the COURAGE in Europe project. The goodness-of-fit of a two-correlated-factors model and the reliability of the scores obtained were assessed. Cross-country invariance was tested employing a multiple group confirmatory factor analysis, through sequential constraint imposition. In each country, measurement invariance was tested across time frames (morning, afternoon and evening) and days of the week (weekday and weekend). The results found support for the hypothesis of a two-correlated-factors (positive and negative affect) structure; the reliability of the positive, the negative and the net affect scores showed appropriate values. A high equivalence across the three national samples was found: all items except one showed strong measurement invariance indicating that respondents from Finland, Poland, and Spain attribute the same meaning to the latent construct under study, and the levels of the underlying items are equal in all three countries. Similar results were found for the measurement equivalence across time frames and days of the week. Our findings support the assumption of comparability across the different samples considered; in general, higher positive affect and lower negative affect were found in Finland, in the evening and at the weekendThe research leading to these results has received funding from the European Community’s Seventh Framework Programme (FP7/2007-2013) under grant agreement number 223071 (COURAGE in Europe), from the Instituto de Salud Carlos III-FIS research grants number PS09/00295 and PS09/01845, and from the Spanish Ministry of Science and Innovation ACI-Promociona (ACI2009-1010). The study was supported by the Instituto de Salud Carlos III, Centro de Investigación Biomédica Red de Salud Mental (CIBERSAM), and the AGES-CM Programme (AGES-S2010/BMD-2422), B.O. is grateful to the Sara Borrell postdoctoral programme (reference no. CD12/00429) supported by the Instituto de Salud Carlos III, Spain

El cielo sobre Jerez. Conservación de las cubiertas y bóvedas de la iglesia de San Miguel

Los desprendimientos puntuales de elementos decorativos de las bóvedas de la nave central y la epístola que tuvieron lugar a finales de 2003 en la Iglesia de San Miguel de Jerez provocaron la intervención con carácter de urgencia de un equipo técnico del IAPH. A lo largo del artículo se describe todo el proceso de investigación e intervención llevado a cabo, en el que tras un exhaustivo estudio multidisciplinar se generó un grado de conocimiento del edificio que permitió elaborar un diagnóstico de su estado de conservación y una propuesta de intervención que, superando las dificultades implícitas a la especificidad de la patología, permitió dar respuesta a la situación de riesgo generada, eliminando las causas de las patologías detectadas, garantizando la seguridad y programando las operaciones de mantenimiento que prolongan la efectividad de las actuaciones de conservación desarrolladas. Proceso que concluyó en enero de 2008 con la finalización de la puesta en obra del Proyecto Básico y de Ejecución de Intervención en la Iglesia de San Miguel de Jerez de la Frontera: reparación de nervaduras y elementos decorativos de las bóvedas, protección de vidrieras y reparación de las cubiertas. Este artículo forma parte de las actividades divulgativas generadas como transferencia del proceso de la intervenciónLos desprendimientos puntuales de elementos decorativos de las bóvedas de la nave central y la epístola que tuvieron lugar a finales de 2003 en la Iglesia de San Miguel de Jerez provocaron la intervención con carácter de urgencia de un equipo técnico del IAPH. A lo largo del artículo se describe todo el proceso de investigación e intervención llevado a cabo, en el que tras un exhaustivo estudio multidisciplinar se generó un grado de conocimiento del edificio que permitió elaborar un diagnóstico de su estado de conservación y una propuesta de intervención que, superando las dificultades implícitas a la especificidad de la patología, permitió dar respuesta a la situación de riesgo generada, eliminando las causas de las patologías detectadas, garantizando la seguridad y programando las operaciones de mantenimiento que prolongan la efectividad de las actuaciones de conservación desarrolladas. Proceso que concluyó en enero de 2008 con la finalización de la puesta en obra del Proyecto Básico y de Ejecución de Intervención en la Iglesia de San Miguel de Jerez de la Frontera: reparación de nervaduras y elementos decorativos de las bóvedas, protección de vidrieras y reparación de las cubiertas. Este artículo forma parte de las actividades divulgativas generadas como transferencia del proceso de la intervención

Erythrocyte Membrane Nanomechanical Rigidity Is Decreased in Obese Patients

This work intends to describe the physical properties of red blood cell (RBC) membranes in obese adults. The hypothesis driving this research is that obesity, in addition to increasing the amount of body fat, will also modify the lipid composition of membranes in cells other than adipocytes. Forty-nine control volunteers (16 male, 33 female, BMI 21.8 ± 5.6 and 21.5 ± 4.2 kg/m2, respectively) and 52 obese subjects (16 male and 36 female, BMI 38.2± 11.0 and 40.7 ± 8.7 kg/m2, respectively) were examined. The two physical techniques applied were atomic force microscopy (AFM) in the force spectroscopy mode, which allows the micromechanical measurement of penetration forces, and fluorescence anisotropy of trimethylammonium diphenylhexatriene (TMA-DPH), which provides information on lipid order at the membrane polar–nonpolar interface. These techniques, in combination with lipidomic studies, revealed a decreased rigidity in the interfacial region of the RBC membranes of obese as compared to control patients, related to parallel changes in lipid composition. Lipidomic data show an increase in the cholesterol/phospholipid mole ratio and a decrease in sphingomyelin contents in obese membranes. ω-3 fatty acids (e.g., docosahexaenoic acid) appear to be less prevalent in obese patient RBCs, and this is the case for both the global fatty acid distribution and for the individual major lipids in the membrane phosphatidylcholine (PC), phosphatidylethanolamine (PE) and phosphatidylserine (PS). Moreover, some ω-6 fatty acids (e.g., arachidonic acid) are increased in obese patient RBCs. The switch from ω-3 to ω-6 lipids in obese subjects could be a major factor explaining the higher interfacial fluidity in obese patient RBC membranes.This work was supported in part by the Basque Government Department of Economic Development, grant No. KK-2019/00028 (OBINTER); the Basque Government Department of Education, grants No. IT1264-19, IT1281-19, IT1270-19, and IT1625-22; the Basque Government Department of Health, grants No. 2019-222030, 2020-333023; Fundación Ramón Areces; and by Centre for the Development of Industrial Technology (CDTI) of the Spanish Ministry of Science and Innovation under the grant agreement: TECNOMIFOOD project (CER-20191010) and Basque Government: IT1625-22

Early mobilization after total hip or knee arthroplasty: a substudy of the POWER.2 study

Background Early mobilization after surgery is a cornerstone of the Enhanced Recovery After Surgery (ERAS) programs in total hip arthroplasty (THA) or total knee arthroplasty (TKA). Our goal was to determine the time to mobilization after this surgery and the factors associated with early mobilization. Methods This was a predefined substudy of the POWER.2 study, a prospective cohort study conducted in patients undergoing THA and TKA at 131 Spanish hospitals. The primary outcome was the time until mobilization after surgery as well as determining those perioperative factors associated with early mobilization after surgery. Results A total of 6093 patients were included. The median time to achieve mobilization after the end of the surgery was 24 hours [16–30]. 4,222 (69.3%) patients moved in ≤ 24 hours after surgery. Local anesthesia [OR = 0.80 (95% confidence interval [CI]: 0.72–0.90); p = 0.001], surgery performed in a self-declared ERAS center [OR = 0.57 (95% CI: 0.55–0.60); p < 0.001], mean adherence to ERAS items [OR = 0.93 (95% CI: 0.92–0.93); p < 0.001], and preoperative hemoglobin [OR = 0.97 (95% CI: 0.96–0.98); p < 0.001] were associated with shorter time to mobilization. Conclusions Most THA and TKA patients mobilize in the first postoperative day, early time to mobilization was associated with the compliance with ERAS protocols, preoperative hemoglobin, and local anesthesia, and with the absence of a urinary catheter, surgical drains, epidural analgesia, and postoperative complications. The perioperative elements that are associated with early mobilization are mostly modifiable, so there is room for improvement

A UHPLC-Mass Spectrometry View of Human Melanocytic Cells Uncovers Potential Lipid Biomarkers of Melanoma

Melanoma is the deadliest form of skin cancer due to its ability to colonize distant sites and initiate metastasis. Although these processes largely depend on the lipid-based cell membrane scaffold, our understanding of the melanoma lipid phenotype lags behind most other aspects of this tumor cell. Here, we examined a panel of normal human epidermal and nevus melanocytes and primary and metastatic melanoma cell lines to determine whether distinctive cell-intrinsic lipidomes can discern non-neoplastic from neoplastic melanocytes and define their metastatic potential. Lipidome profiles were obtained by UHPLC-ESI mass-spectrometry, and differences in the signatures were analyzed by multivariate statistical analyses. Significant and highly specific changes in more than 30 lipid species were annotated in the initiation of melanoma, whereas less numerous changes were associated with melanoma progression and the non-malignant transformation of nevus melanocytes. Notably, the “malignancy lipid signature” features marked drops in pivotal membrane lipids, like sphingomyelins, and aberrant elevation of ether-type lipids and phosphatidylglycerol and phosphatidylinositol variants, suggesting a previously undefined remodeling of sphingolipid and glycerophospholipid metabolism. Besides broadening the molecular definition of this neoplasm, the different lipid profiles identified may help improve the clinical diagnosis/prognosis and facilitate therapeutic interventions for cutaneous melanoma.This research was funded in part by grants from the Ministry of Economy; Industry and Competitiveness (RTC-2015-3693-1); Ministry of Science and Innovation (RTI-2018-095134-B-I00); Basque Government (IT971-16; IT1162-19; KK2016-036; KK2017-041 and KK2018-00090) and UPV/EHU (GIU17/066)