CXCL12-induced neurotoxicity critically depends on NMDA receptor-gated and L-type Ca2+ channels upstream of p38 MAPK.

BackgroundThe chemokine receptor CXCR4 (CD184) and its natural ligand CXCL12 contribute to many physiological processes, including decisions about cell death and survival in the central nervous system. In addition, CXCR4 is a co-receptor for human immunodeficiency virus (HIV)-1 and mediates the neurotoxicity of the viral envelope protein gp120. However, we previously observed that CXCL12 also causes toxicity in cerebrocortical neurons but the cellular mechanism remained incompletely defined.MethodsPrimary neuronal-glial cerebrocortical cell cultures from rat were exposed to a neurotoxicity-inducing CXCL12 concentration for different times and the activity of the stress-associated mitogen-activated protein kinase p38 (p38 MAPK) was assessed using an in vitro kinase assay. Neurotoxicity of CXCL12 and cellular localization of p38 MAPK was analyzed by immunofluorescence microscopy. Pharmacological inhibition of NMDA-type glutamate receptor-gated ion channels (NMDAR) of L-type Ca2+ channels was employed during 12- and 24-h exposure to neurotoxic amounts of CXCL12 to study the effects on active p38 MAPK and neuronal survival by Western blotting and microscopy, respectively. Neurotoxicity of CXCL12 was also assessed during pharmacological inhibition of p38 MAPK.ResultsHere, we show that a neurotoxic amount of CXCL12 triggers a significant increase of endogenous p38 MAPK activity in cerebrocortical cells. Immunofluorescence and Western blotting experiments with mixed neuronal-glial and neuron-depleted glial cerebrocortical cells revealed that the majority of active/phosphorylated p38 MAPK was located in neurons. Blockade of NMDAR-gated ion channels or L-type Ca2+ channels both abrogated an increase of active p38 MAPK and toxicity of CXCL12 in cerebrocortical neurons. Inhibition of L-type Ca2+ channels with nimodipine kept the active kinase at levels not significantly different from baseline while blocking NMDAR with MK-801 strongly reduced phosphorylated p38 MAPK below baseline. Finally, we confirmed that directly blocking p38 MAPK also abrogated neurotoxicity of CXCL12.ConclusionsOur findings link CXCL12-induced neuronal death to the regulation of NMDAR-gated ion channels and L-type Ca2+ channels upstream of p38 MAPK activation

Innovative Tourism Clusters: Myth or Reality? Empirical Evidence from Benidorm

Paper submitted to TCVT3 Bozen/Bolzano, 10-12 April 2014, International workshop on Tourists as Consumers, Visitors, TravellersThis paper analyses the applicability of the cluster concept to tourist destinations, taking Benidorm as the reference for study: a destination on the Mediterranean coast of Spain which has attained a high degree of competitive success with the sun and sand product in which it has specialized. The main result obtained is that Benidorm's success is not derived from or favoured by the existence of a cluster in the destination, since important elements are present that prevent affirmation of its existence. This paper is the preliminary result of a research project that has been carried out within the framework of the project "Methodology, criteria and impementation of the cluster theory in consolidated tourism areas: innovation, competitiveness and territorial synergies" under the Spanish National R&D&I Plan 2008-2011 supported by the Ministry of Science and Innovation.This research has been carried out within the framework of the project “Methodology, criteria and implementation of the cluster theory in consolidated tourism areas: innovation, competitiveness and territorial synergies” under the Spanish National R&D&I Plan 2008-2011 supported by the Ministry of Science and Innovation

Consumer behavior in the digital age

In recent decades, the Internet, evolving technologies, and social media have led to the evolution of consumer behavior. The changes in customer behavior driven by digital developments provide many opportunities and challenges that businesses also need to deal with online. The better companies know about the behavior of their customers, the easier they can engage with them using strategies such as content marketing, User Experience (UX), influencers marketing, User-Generated Content (UGC), or Electronic Word of Mouth (eWOM). These strategies are essential to get more sales and to develop businesses online, as such strategies increase the engagement with users and influence their behavior. This Special Edition of JOSD focuses on the analysis of consumer behavior in the digital age and, by doing so, contributes to extant knowledge about digital marketing strategies, online consumer behavior, and new digital business models such as mobile applications or shared economy.FCT- Foundation for Science and TechnologyPortuguese Foundation for Science and Technology [UIDB/04020/2020]info:eu-repo/semantics/publishedVersio

Urinary cyclophilin A as marker of tubular cell death and kidney injury

Background: Despite the term acute kidney injury (AKI), clinical biomarkers for AKI re-flect function rather than injury and independent markers of injury are needed. Tubular cell death, including necroptotic cell death, is a key feature of AKI. Cyclophilin A (CypA) is an intracellular protein that has been reported to be released during necroptosis. We have now explored CypA as a potential marker for kidney injury in cultured tubular cells and in clinical settings of ischemia-reperfusion injury (IRI), characterized by limitations of current diagnostic criteria for AKI. Meth-ods: CypA was analyzed in cultured human and murine proximal tubular epithelial cells exposed to chemical hypoxia, hypoxia/reoxygenation (H/R) or other cell death (apoptosis, necroptosis, fer-roptosis) inducers. Urinary levels of CypA (uCypA) were analyzed in patients after nephron sparing surgery (NSS) in which the contralateral kidney is not disturbed and kidney grafts with initial function. Results: Intracellular CypA remained unchanged while supernatant CypA increased in parallel to cell death induction. uCypA levels were higher in NSS patients with renal artery clamping (that is, with NSS-IRI) than in no clamping (NSS-no IRI), and in kidney transplantation (KT) recipients (KT-IRI) even in the presence of preserved or improving kidney function, while this was not the case for urinary Neutrophil gelatinase-associated lipocalin (NGAL). Furthermore, higher uCypA levels in NSS patients were associated with longer surgery duration and the incidence of AKI increased from 10% when using serum creatinine (sCr) or urinary output criteria to 36% when using high uCypA levels in NNS clamping patients. Conclusions: CypA is released by kidney tubular cells during different forms of cell death, and uCypA increased during IRI-induced clinical kidney injury independently from kidney function parameters. Thus, uCypA is a potential bi-omarker of kidney injury, which is independent from decreased kidney functionResearch by the authors was funded by FIS/ FEDER funds (PI17/00257, PI18/01386, PI19/00588, PI19/00815, DTS18/00032, ERA-PerMed-JTC2018 (KIDNEY ATTACK AC18/00064 and PERSTIGAN AC18/00071, ISCIII-RETIC REDinREN RD016/0009), Sociedad Española de Nefrología, FRIAT, Comunidad de Madrid en Biomedicina B2017/BMD-3686 CIFRA2-CM. Salary support: ISCIII Miguel Servet to A.B.S., MICIN Ramon y Cajal to M.D.S.-N., REDinREN RD016/0009 to M.F.-B.,SENEFRO to D.M.-S. and Consejería de Educación, Juventud y Deporte (Comunidad de Madrid/FSE) to A.M.L.-

Molecular pathways driving omeprazole nephrotoxicity

Omeprazole, a proton pump inhibitor used to treat peptic ulcer and gastroesophageal reflux disease, has been associated to chronic kidney disease and acute interstitial nephritis. However, whether omeprazole is toxic to renal cells is unknown. Omeprazole has a lethal effect over some cancer cells, and cell death is a key process in kidney disease. Thus, we evaluated the potential lethal effect of omeprazole over tubular cells. Omeprazole induced dose-dependent cell death in human and murine proximal tubular cell lines and in human primary proximal tubular cell cultures. Increased cell death was observed at the high concentrations used in cancer cell studies and also at lower concentrations similar to those in peptic ulcer patient serum. Cell death induced by omeprazole had features of necrosis such as annexin V/7-AAD staining, LDH release, vacuolization and irregular chromatin condensation. Weak activation of caspase-3 was observed but inhibitors of caspases (zVAD), necroptosis (Necrostatin-1) or ferroptosis (Ferrostatin-1) did not prevent omeprazole-induced death. However, omeprazole promoted a strong oxidative stress response affecting mitochondria and lysosomes and the antioxidant N-acetyl-cysteine reduced oxidative stress and cell death. By contrast, iron overload increased cell death. An adaptive increase in the antiapoptotic protein BclxL failed to protect cells. In mice, parenteral omeprazole increased tubular cell death and the expression of NGAL and HO-1, markers of renal injury and oxidative stress, respectively. In conclusion, omeprazole nephrotoxicity may be related to induction of oxidative stress and renal tubular cell deathSupported by FIS CP12/03262, CP14/00133, PI16/02057, PI16/ 01900, PI18/01366, PI19/00588, PI19/00815, DTS18/00032, ERAPerMed-JTC2018 (KIDNEY ATTACK AC18/00064 and PERSTIGAN AC18/00071, ISCIII-RETIC REDinREN RD016/0009 FEDER funds, Sociedad Española de Nefrología, Fundacion Renal Iñigo Álvarez de Toledo (FRIAT), ISCIII Miguel Servet (ABS, MDS-N), ISCIII Sara Borrell (JMM-M), Comunidad de Madrid CIFRA2 B2017/BMD-3686 (MF-B and DM-S

Development of lactoferrin-loaded liposomes for the management of dry eye disease and ocular inflammation

Dry eye disease (DED) is a high prevalent multifactorial disease characterized by a lack of homeostasis of the tear film which causes ocular surface inflammation, soreness, and visual disturbance. Conventional ophthalmic treatments present limitations such as low bioavailability and side effects. Lactoferrin (LF) constitutes a promising therapeutic tool, but its poor aqueous stability and high nasolacrimal duct drainage hinder its potential efficacy. In this study, we incorporate lactoferrin into hyaluronic acid coated liposomes by the lipid film method, followed by high pressure homogenization. Pharmacokinetic and pharmacodynamic profiles were evaluated in vitro and ex vivo. Cytotoxicity and ocular tolerance were assayed both in vitro and in vivo using New Zealand rabbits, as well as dry eye and anti-inflammatory treatments. LF loaded liposomes showed an average size of 90 nm, monomodal population, positive surface charge and a high molecular weight protein encapsulation of 53%. Biopharmaceutical behaviour was enhanced by the nanocarrier, and any cytotoxic effect was studied in human corneal epithelial cells. Developed liposomes revealed the ability to reverse dry eye symptoms and possess anti-inflammatory efficacy, without inducing ocular irritation. Hence, lactoferrin loaded liposomes could offer an innovative nanotechnological tool as suitable approach in the treatment of DED.FCT -Fundação para a Ciência e a Tecnologia(2017SGR1477)info:eu-repo/semantics/publishedVersio

PELFI Project: Recruitment and Sociodemographic Characteristics of Immigrant and Autochthonous Families from Alicante and Barcelona City Subcohorts

Este artículo corresponde al “Proyecto de Estudios Longitudinales de Familias Inmigradas (PELFI)” del Subprograma de Inmigración y Salud del CIBERESP y describe el trabajo de campo basal y principales características socio-demográficas de dos sub-cohortes de familias inmigrantes y autóctonas. El diseño es observacional prospectivo. La población de estudio se definió como una muestra no probabilística de 180 familias de origen colombiano, ecuatoriano y marroquí y 50 españolas. Se entrevistó a 473 personas adultas entre 18 y 65 años (59,8% mujeres, 68,5% ocupados/as) y a 304 adolescentes entre 12 y17 años (53,9% mujeres, 27,1% nacidos en España pero de padres inmigrados) de cada familia, mediante dos cuestionarios diseñados ad hoc. La tasa de cooperación fue del 82,0% con una velocidad media de reclutamiento de 1,3 familias diarias. En total, se reclutó a 250 familias, 82 procedentes de Ecuador, 82 de Colombia, 29 de Marruecos y 57 españolas. Los adultos inmigrados llevaban una media de 13 años en España. La combinación de técnicas no probabilísticas permitió el acceso y velocidad de reclutamiento. Este estudio aporta información clave para el diseño y mejora de este tipo de cohortes en familias inmigradas.This study is a part of the multi-centre project “Platform of Longitudinal Studies of Immigrant Families (PELFI)” of the Immigration and Health Subprogram of the CIBER-ESP. It describes the field work and data collection of two sub-cohorts of immigrant and native families, and their main socio-demographic characteristics. Prospective observational cohort study in carried out in Barcelona and Alicante, Spain. The study population is a non-probabilistic sample of 180 families of Colombian, Ecuadorian and Moroccan origin and 50 families of Spanish origin. We interviewed adults aged 18-65 years and adolescents aged 12-17 years in each family, through two questionnaires (adolescent/adult). The cooperation rate was 82.0% with an average recruitment rate of 1.3 families per day. In total, 250 families have been recruited, 82 from Ecuador, 82 from Colombia, 29 from Morocco and 57 from Spain. A total of 473 adults (59.8% women and 68.5% employed) were surveyed. Immigrant adults have an average of 13 years living in Spain. A total of 304 adolescents (53.9% female, 27.1% born in Spain but with immigrant parents) were surveyed. The combination of non-probabilistic techniques promoted access and improved recruitment speed. This study provides key information for the design and improvement of cohort studies with immigrant families.Proyectos Fondo Investigación Sanitaria números PI14/01146 y PI14/02005 e Instituto de Salud Carlos III-FEDER

Mono- and dicationic DABCO/Quinuclidine composed nanomaterials for the loading of steroidal drug: 32 factorial design and physicochemical characterization

Oil-in-water nanoemulsions (NEs) are considered a suitable nanotechnological approach to improve the eye-related bioavailability of lipophilic drugs. The potential of cationic NEs is prominent due to the electrostatic interaction that occurs between the positively charged droplets with the negatively charged mucins present in the tear film. This interaction offers prolonged NEs residence at the ocular surface, increasing the drug absorption. Triamcinolone acetonide (TA) is one of the first pharmacologic strategies applied as an intravitreal injection in the treatment of age-related macular degeneration (AMD). Newly synthesized quaternary derivatives of 1,4-diazabicyclo[2.2.2]octane (DABCO) and quinuclidine surfactants have been screened with the purpose to select the best compound to formulate long-term stable NEs that combine the best physicochemical properties for the loading of TA intended for ocular administration.This work was funded by the Portuguese Science and Technology Foundation (FCT) from the Ministry of Science and Technology (MCTES), European Social Fund (FSE) of EU, for the scholar ship SFRH/BD/130555/2017 granted to A. R. Fernandes, and for the projects UIDB/04469/2020 (CEB strategic fund) and UIDB/04033/2020 (CITAB), co-funded by European Funds (PRODER/COMPETE) and FEDER, under the Partnership Agreement PT2020.info:eu-repo/semantics/publishedVersio