Vascular lysyl oxidase over-expression alters extracellular matrix structure and induces oxidative stress

Lysyl oxidase (LOX) participates in the assembly of collagen and elastin fibres. The impact of vascular LOX over-expression on extracellular matrix (ECM) structure and its contribution to oxidative stress has been analysed. Methods Studies were conducted on mice over-expressing LOX (Tg), specifically in smooth muscle cells (VSMC). Gene expression was assessed by real-time PCR analysis. Sirius Red staining, H 2 O 2 production and NADPH oxidase activity were analysed in different vascular beds. The size and number of fenestra of the internal elastic lamina were determined by confocal microscopy. Results LOX activity was up-regulated in VSMC of transgenic mice compared with cells from control animals. At the same time, transgenic cells deposited more organised elastin fibres and their supernatants induced a stronger collagen assembly in in vitro assays. Vascular collagen cross-linking was also higher in Tg mice, which showed a decrease in the size of fenestrae and an enhanced expression of Fibulin-5. Interestingly, higher H 2 O 2 production and NADPH oxidase activity was detected in the vascular wall from transgenic mice. The H 2 O 2 scavenger catalase attenuated the stronger deposition of mature elastin fibres induced by LOX transgenesis. Conclusions LOX over-expression in VSMC was associated with a change in the structure of collagen and elastin fibres. LOX could constitute a novel source of oxidative stress that might participate in elastin changes and contribute to vascular remodellingLa lisil oxidasa (LOX) contribuye al ensamblaje de las fibras de colágeno y elastina de la matriz extracelular (MEC). Hemos determinado las consecuencias de la sobre-expresión vascular de LOX sobre la estructura de la MEC y su contribución al estrés oxidativo. Métodos: Los estudios se desarrollaron en ratones que sobre-expresan la LOX (Tg) específicamente en células musculares lisas vasculares (CMLV). Se realizaron análisis por PCR a tiempo real, tinción de rojo sirio, producción de H2O2 y actividad NADPH oxidasa. Se caracterizaron las fenestras de la lámina elástica interna mediante microscopía confocal. Resultados: Las CMLV de ratones transgénicos presentaron niveles de actividad LOX superiores a los de animales control. En consonancia, las células transgénicas depositaron más fibras de elastina organizada y sus sobrenadantes indujeron un mayor ensamblaje de colágeno en ensayos in vitro. El nivel de colágeno maduro fue superior en la pared vascular de ratones Tg, que presentaban un menor área de las fenestras y un aumento de la expresión de la Fibulina-5. La producción vascular de H2O2 y la actividad NADPH oxidasa fueron superiores en los ratones transgénicos. La incubación de CMLV con catalasa atenuó el incremento en la deposición de fibras de elastina madura inducido por la transgénesis de LOX. Conclusiones: La sobre-expresión de la LOX en CMLV se asocia a una alteración de la estructura vascular del colágeno y la elastina. La LOX podría constituir una nueva fuente de estrés oxidativo que participaría en la alteración estructural de la MEC y podría contribuir al remodelado vascularEste estudio se ha financiado por la Fundación Española de Aterosclerosis, Beca SEA/FEA de Investigación básica 2016 y por el Ministerio de Economía y Competitividad (MINECO)-Instituto de Salud Carlos III (ISCIII) [proyectos PI15/01016, PI13/01488, SAF2012-36400; SAF2015-64767-R]. El CIBER de Enfermedades Cardiovasculares es una iniciativa del ISCIII. AMB recibió una ayuda del programa Ramón y Cajal (RYC-2010-06473). El estudio ha sido cofinanciado por el Fondo Europeo de Desarrollo Regional (FEDER

C-Src, ERK1/2 and Rho kinasemediate hydrogen peroxide-induced vascular contraction in hypertension: Role ofTXA2, NAD(P)H oxidase andmitochondria

AIM: : The aim of this study was to analyse the signalling pathways involved in H2O2 vascular responses in hypertension. METHODS: Vascular function, thromboxane A2 (TXA2) production, oxidative stress and protein expression were determined in mesenteric resistance arteries (MRAs) from hypertensive (spontaneously hypertensive rats, SHR) and normotensive Wistar Kyoto (WKY) rats. RESULTS: H2O2 and the TP agonist U46619 induced greater contractile responses in MRA from SHR than WKY. Moreover, H2O2 increased TXA2 production more in SHR than in WKY. The c-Src inhibitor PP1 reduced H2O2 and U46619-induced contraction and TXA2 release in both strains. The ERK1/2 inhibitor PD98059 reduced H2O2 but not U46619-induced contraction only in SHR arteries. The Rho kinase inhibitor Y26372 reduced H2O2 and U46619-induced contractions only in SHR arteries. Basal c-Src, ERK1/2 and Rho kinase expression were greater in MRA from SHR than WKY. In SHR, the combination of PD98059 with the TP antagonist SQ29548 but not with Y27632 inhibited the H2O2 contraction more than each inhibitor alone. H2O2 and U46619 increased NAD(P)H oxidase activity and O2 production and decreased mitochondrial membrane potential in vessels from SHR. The effects induced by H2O2 were abolished by inhibitors of TXA2 synthase, ERK1/2 and c-Src. The mitochondrial antioxidant mitoTEMPO reduced H2O2-induced contraction and NAD(P)H oxidase activation. CONCLUSION: In arteries from WKY, c-Src mediates H2O2 contractile responses by modulating TXA2 release and TXA2 effect. In SHR, H2O2 induces c-Src dependent TXA2 release that provokes vascular contractile responses through Rho kinase, c-Src and O2 from NAD(P)H Oxidase and mitochondria. Moreover, ERK1/2 activation contributes to H2O2 contraction in SHR through effects on mitochondria/NAD(P)H Oxidase

Síntomas de depresión en los adolescentes, estrategias de solución de problemas y educación para la salud comunitaria

The aims of this investigation have been the study of the presence of depressive symptomatology and their relationship with cognitive distortions in adolescent scholars. Children's Depression Inventory (QDL Kovacs, 1992), Peer Nomination Inventory of Depression (PNID, Leflcowitz y Tesiny, 1981) and Children's Cognitive Distortions Questionnaire (CDCN-1, Bas, 1987) have been administered on a representative sample of 908 Sevillian adolescents (aged 12 to 16). The results revealed an 1178% of depressive symptomatology with an upward lineal tendency regarding the age and with superior scores in the case of women. The number of cognitive distortions has been related positively with the presence of this symptomatology. This point shows to be one of the main predictors of the punctuations in CDL The depressive symptomatology keeps relation to the most important moments in the youth's cognitive development. An increase of cognitive distortions has been appreciated on aged 13. Later on, this one stays constant. CDCN-1 is considered as an appropriate complement of CDI, on the contrary that PNID, probably for the process of intemalization of depressive manifestations. You cannot grant a causal role on depression to the cognitive biases. It is suggested that the coincidence between CDI and CDCN-1 about raised scores is a sign of a high risk (vulnerability) to develop a first depressive episode.Para determinar la prevalencia de los síntomas de depresión y examinar su relación con las estrategias de solución de problemas en adolescentes, se administraron dos cuestionarios a 118 estudiantes de bachillerato (14-15 años) en el distrito de Tetuán (Madrid). El estudio de los síntomas de prevalencia y los estilos y estrategias de afrontamiento se realizó mediante la autoaplicación del Inventario de Depresión de Beck y el Cuestionario de Situaciones de Afrontamiento (CASQ) de Seiffge-Krenke. Los datos muestran que las estrategias de afrontamiento disfuncionales para la solución de problemas eran más utilizadas entre los adolescentes con más síntomas de depresión. Es necesario analizar estos resultados en el contexto de planificar programas de educación para la salud comunitaria que son fundamentales para prevenir la depresión en los adolescentes

Stress compensation by gap monolayers for stacked InAs/GaAs quantum dots solar cells

In this work we report the stacking of 10 and 50 InAs quantum dots layers using 2 monolayers of GaP for stress compensation and a stack period of 18 nm on GaAs (001) substrates. Very good structural and optical quality is found in both samples. Vertical alignment of the dots is observed by transmission electron microscopy suggesting the existence of residual stress around them. Photocurrent measurements show light absorption up to 1.2 μm in the nanostructures together with a reduction in the blue response of the device. As a result of the phosphorus incorporation in the barriers, a very high thermal activation energy (431 meV) has also been obtained for the quantum dot emission

Structure and magnetism of single-phase epitaxial γ′-Fe4N

Single phase epitaxial pure γ′-Fe4N films are grown on MgO (001) by molecular beam epitaxy of iron in the presence of nitrogen obtained from a radio frequency atomic source. The epitaxial, single phase nature of the films is revealed by x-ray diffraction and by the local magnetic environment investigated by Mössbauer spectroscopy. The macroscopic magnetic properties of the γ′-Fe4N films are studied in detail by means of transverse Kerr effect measurements. The hysteresis loops are consistent with the cubic atomic structure, displaying easy [100] magnetization directions. The films are single domain at remanence, and the reversal is dominated by 180° or 90° domain wall nucleation and propagation, depending on the applied field direction. When 90° domain walls are responsible for the magnetization reversal, this proceeds in two stages, and the measured coercive fields vary accordingly. Magnetic domain observations reveal the two distinct reversal —driven by 180° or 90° domain walls— modes displaying large domains, of the order of mm. From magnetometer techniques, the saturation magnetization, μ0Ms, is measured to be 1.8 T. A magneto-optical torque technique is used to obtain a value of the anisotropy constant of 2.9×104J/m3.The authors acknowledge partial financing from EC project HIDEMAR G5RD-CT-2002-00731 and PHANTOMS network. The authors are indebted to A. Gupta and K. V. Rao from the department of Materials Science and Engineering, KTH, Sweden for help with the low T SQUID measurements, and to L. Ballcels and M. A. García from Materials Science ICMM CSIC, Spain for high-T VSM measurements. This work was part of the research program of the Foundation for Fundamental Research on Matter-FOM, The Netherlands. J.M.G.M. acknowledges financing through the Ramón y Cajal program from the Spanish MCyT.Peer reviewe

Effects of ammonium nitrate, cesium chloride and tetraethylammonium on high-affinity potassium uptake in habanero pepper plantlets (Capsicum chinense Jacq.)

Potassium (K+) is an essential nutrient and the most abundant cation in plant cells. Plants have a wide variety of transport systems for K+ acquisition that catalyze K+ uptake across a wide spectrum of external K+ concentrations and mediate K+ movement within the plant, as well as its release into the environment. The KUP/HAK/KT transporter family plays a key role in K+ homeostasis in plant cells. The present study demonstrates that habanero pepper plantlets have a clear pattern of K+ uptake when resupplemented with K+ after K+ starvation. Habanero pepper plantlets, re-supplemented with a solution containing low concentrations of K+ after 72, 96 or 120 h of K+ starvation were able to decrease the amount of K+ in the solution at different time points. To study the effect of NH4+, we added different concentrations of NH4NO3 to the medium solution and demonstrated that NH4+ inhibited K+ uptake in a dose-dependent manner. When the plantlets were subjected to K+ starvation for 72 h and then resupplemented with 50 or 100 μM K+, exposure to K+ channel blockers (10 mM CsCl and 20 mM TEA) decreased their K+ uptake compared with the control treatment. A model demonstrating the process of K+ uptake through an NH4+-insensitive component was proposed.Key words: Potassium, high affinity transporters, channel blockers, ammonium

Toll-like receptor 4 contributes to vascular remodelling and endothelial dysfunction in angiotensin II-induced hypertension

This is the peer-reviewed version of the following article: "Toll-like receptor 4 contributes to vascular remodelling and endothelial dysfunction in angiotensin II-induced hypertension", British Journal of Pharmacology 172.12 (2015): 3159-76 which has been published in final form at http://dx.doi.org/10.1111/bph.13117 This article may be used for non-commercial purposes in accordance with Wiley-VCH Terms and Conditions for Self-ArchivingBackground and Purpose Toll-like receptor 4 (TLR4) signalling contributes to inflammatory cardiovascular diseases, but its role in hypertension and the associated vascular damage is not known. We investigated whether TLR4 activation contributed to angiotensin II (AngII)-induced hypertension and the associated vascular structural, mechanical and functional alterations. Experimental Approach AngII was infused (1.44 mg·kg−1·day−1, s.c.) for 2 weeks in C57BL6 mice, treated with a neutralizing anti-TLR4 antibody or IgG (1 μg·day−1); systolic BP (SBP) and aortic cytokine levels were measured. Structural, mechanical and contractile properties of aortic and mesenteric arterial segments were measured with myography and histology. RT-PCR and Western blotting were used to analyse these tissues and cultured vascular smooth muscle cells (VSMC) from hypertensive rats (SHR). Key Results Aortic TLR4 mRNA levels were raised by AngII infusion. Anti-TLR4 antibody treatment of AngII-treated mice normalised: (i) increased SBP and TNF-α, IL-6 and CCL2 levels; (ii) vascular structural and mechanical changes; (iii) altered aortic phenylephrine- and ACh-induced responses; (iv) increased NOX-1 mRNA levels, superoxide anion production and NAD(P)H oxidase activity and effects of catalase, apocynin, ML-171 and Mito-TEMPO on vascular responses; and (v) reduced NO release and effects of L-NAME on phenylephrine-induced contraction. In VSMC, the MyD88 inhibitor ST-2825 reduced AngII-induced NAD(P)H oxidase activity. The TLR4 inhibitor CLI-095 reduced AngII-induced increased phospho-JNK1/2 and p65 NF-κB subunit nuclear protein expression. Conclusions and Implications TLR4 up-regulation by AngII contributed to the inflammation, endothelial dysfunction, vascular remodelling and stiffness associated with hypertension by mechanisms involving oxidative stress. MyD88-dependent activation and JNK/NF-κB signalling pathways participated in these alterationsThis work was supported by Ministerio de Economía y Competitividad (SAF2012-36400), Instituto de Salud Carlos III (Red de Investigación Cardiovascular RD12/0042/0024 and RD12/0042/0033) and URJC (PRIN13_CS12). AMB was supported by the Ramón y Cajal Program (RYC-2010-06473)