Predicting the onset and persistence of episodes of depression in primary health care. The predictD-Spain study: Methodology

Background: The effects of putative risk factors on the onset and/or persistence of depression remain unclear. We aim to develop comprehensive models to predict the onset and persistence of episodes of depression in primary care. Here we explain the general methodology of the predictD-Spain study and evaluate the reliability of the questionnaires used. Methods: This is a prospective cohort study. A systematic random sample of general practice attendees aged 18 to 75 has been recruited in seven Spanish provinces. Depression is being measured with the CIDI at baseline, and at 6, 12, 24 and 36 months. A set of individual, environmental, genetic, professional and organizational risk factors are to be assessed at each follow-up point. In a separate reliability study, a proportional random sample of 401 participants completed the test-retest (251 researcher-administered and 150 self-administered) between October 2005 and February 2006. We have also checked 118,398 items for data entry from a random sample of 480 patients stratified by province. Results: All items and questionnaires had good test-retest reliability for both methods of administration, except for the use of recreational drugs over the previous six months. Cronbach's alphas were good and their factorial analyses coherent for the three scales evaluated (social support from family and friends, dissatisfaction with paid work, and dissatisfaction with unpaid work). There were 191 (0.16%) data entry errors. Conclusion: The items and questionnaires were reliable and data quality control was excellent. When we eventually obtain our risk index for the onset and persistence of depression, we will be able to determine the individual risk of each patient evaluated in primary health care.The research in Spain was funded by grants from the Spanish Ministry of Health (grant FIS references: PI04/1980, PI0/41771, PI04/2450, and PI06/1442), Andalusian Council of Health (grant references: 05/403, 06/278 and 08/0194), and the Spanish Ministry of Education and Science (grant reference SAF 2006/07192). The Malaga sample, as part of the predictD-International study, was also funded by a grant from The European Commission (reference QL4-CT2002-00683)

Tunable from blue to red emissive composites and solids of silver diphosphane systems with higher quantum yields than the diphosphane ligands

PMMA composites and solids of complexes of formulas [AgX(P–P)]n [n = 1 and 2; X = Cl, NO3, ClO4, CF3COO, and OTf; P–P = dppb, xantphos, (PR2)2C2B10H10 (R = Ph and iPr)] display the whole palette of colors from blue to red upon selection of the anionic ligand (X) and the diphosphane (P–P). The diphosphane seems to play the most important role in tuning the emission energy and thermally activated delayed fluorescence (TADF) behavior. The PMMA composites of the complexes exhibit higher quantum yields than that of the diphosphane ligands and those with dppb are between 28 and 53%. Remarkably, instead of blue-green emissions which dominate the luminescence of silver diphosphane complexes in rigid phases, those with carborane diphosphanes are yellow-orange or orange-red emitters. Theoretical studies have been carried out for complexes with P–P = dppb, X = Cl; P–P = dppic, X = NO3; P–P = dppcc, X = Cl, NO3, and OTf and the mononuclear complexes [AgX(xantphos)] (X = Cl, Br). Optimization of the first excited triplet state was only possible for [AgX(xantphos)] (X = Cl and Br). A mixed MLCT and MC nature could be attributed to the S0 → T1 transition in these three-coordinated complexes.We thank the Ministerio de Economía y Competitividad. (CTQ2016-75816-C2-1-P), PID2019-104379RB-C21 and DGA-FSE (E07_20R) for financial support. We also thank Dr. Rafael Cases Andreu from Departamento de Fisica de la Materia Condensada, Facultad de Ciencias and Instituto de Ciencia de Materiales de Aragón (ICMA), and CSIC–Universidad de Zaragoza (Spain), for fruitful discussions regarding to lifetime measurements.Peer reviewe

Evaluación y seguimiento en parvulario y ciclo inicial : pautas de observación

Este libro es el resultado de un trabajo de intervención y de elaboración, que, a lo largo de casi tres años, han llevado a cabo los miembros de los Servicios Municipales de Asesoramiento Psicopedagógico de Sant Boi de Llobregat y Sant Just Desvern. En los dos primeros capítulos se tratan tres conceptos básicos del proceso educativo (evaluación, seguimiento y observación), proponiéndose ubicarlos en el contexto más global de la planificación y conducción del proceso de enseñanza-aprendizaje, destacando sus conexiones mutuas y su complementariedad, así como las funciones que, pueden y deben desempeñar. En el tercer capítulo se recogen íntegramente las pautas de observación, precedidas de unos comentarios que intentan explicar su estructura y los enfoques adoptados en cada caso. En el capítulo cuarto se aborda la cuestión de cómo sistematizar las observaciones durante períodos de tiempo más o menos amplios.En el quinto capítulo se exponen las líneas generales de utilización de las pautas y algunas utilizaciones reales de las mismas. Es un libro útil para introducir en la práctica educativa el método de observación en la evaluación de educación infantil y primer ciclo de primaria.CataluñaBiblioteca de Educación del Ministerio de Educación, Cultura y Deporte; Calle San Agustín, 5; 28014 Madrid; Tel. +34917748000; [email protected]

Tungstate-targeting of BKαβ1 channels tunes ERK phosphorylation and cell proliferation in human vascular smooth muscle

Despite the substantial knowledge on the antidiabetic, antiobesity and antihypertensive actions of tungstate, information on its primary target/s is scarce. Tungstate activates both the ERK1/2 pathway and the vascular voltage- and Ca2+-dependent large-conductance BKαβ1 potassium channel, which modulates vascular smooth muscle cell (VSMC) proliferation and function, respectively. Here, we have assessed the possible involvement of BKαβ1 channels in the tungstate-induced ERK phosphorylation and its relevance for VSMC proliferation. Western blot analysis in HEK cell lines showed that expression of vascular BKαβ1 channels potentiates the tungstate-induced ERK1/2 phosphorylation in a Gi/o protein-dependent manner. Tungstate activated BKαβ1 channels upstream of G proteins as channel activation was not altered by the inhibition of G proteins with GDPβS or pertussis toxin. Moreover, analysis of Gi/o protein activation measuring the FRET among heterologously expressed Gi protein subunits suggested that tungstate-targeting of BKαβ1 channels promotes G protein activation. Single channel recordings on VSMCs from wild-type and β1-knockout mice indicated that the presence of the regulatory β1 subunit was essential for the tungstate-mediated activation of BK channels in VSMCs. Moreover, the specific BK channel blocker iberiotoxin lowered tungstate-induced ERK phosphorylation by 55% and partially reverted (by 51%) the tungstate-produced reduction of platelet-derived growth factor (PDGF)-induced proliferation in human VSMCs. Our observations indicate that tungstate-targeting of BKαβ1 channels promotes activation of PTX-sensitive Gi proteins to enhance the tungstate-induced phosphorylation of ERK, and inhibits PDGF-stimulated cell proliferation in human vascular smooth muscle.This work was supported by grants from the Spanish Ministry of Economy and Competitiveness (SAF2012-31089 to JMFF, SAF2012-38140 to MAV, BFU2013-45867-R to JRLL), FEDER Funds, Ministry of Science and Innovation (BFU 2008-00769 to JG, BFU2010-15898 to MTPG), Instituto de Salud Carlos III (RIC RD12/0042/0006, RD12/0042/0014, Red HERACLES) and Junta de Castilla y León (VA094A11-2 to JRLL) and European Community (FP7-People-CIG-321721 to RK)

Tungstate-induced increase in the open probability of BK channels endogenously expressed in freshly isolated vascular myocytes requires the BK channel β₁ subunit.

<p>(A) Representative recordings obtained from inside-out patches clamped at + 60mV from wild-type (WT) and β₁- knockout freshly isolated mouse vascular myocytes, before (control) and after exposure to 1 mM tungstate (1 mM WO₄^2-), as stated. Arrows indicates the closed state level. (B) Average changes (in %) of BK channel open probability (NP_o) induced by 1 mM tungstate on WT and β₁- knockout mouse vascular myocytes at the indicated voltage membrane. Significant differences were found among WT (n = 7) and β₁- knockout (n = 9), P < 0.001 (Student’s t-test). Further depolarization to +80 mV of β₁- knockout inside-out patches did not change the response to tungstate found at +60 mV.</p

Tungstate-induced activation of heterologously expressed Gi/o proteins is mediated by BKαβ₁ channels.

<p>(A) Example of the reinforced membrane fluorescence pattern and emission levels from CFP channel (up-left), YFP (FRET channel) (up-right) and the merge channels (bottom-left) from HEK293 heterologously expressing Gα_i-YFP and CFP-Gβ. Fluorescence microscopy images were recorded by using confocal microscopy (for more details see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0118148#sec002" target="_blank">Methods</a>). FRET signal was determined by using donor ratiometric parameters (458/514) after excitation in the CFP frequency and registering in the YFP emission frequency. (B) Representative FRET changes from HEK293, HEKα or HEKαβ₁ cells transfected with the cDNAs of G protein subunits, in response to 1 mM tungstate (either in the absence or presence of IbTX), as indicated. (C), Average FRET changes for the different experimental conditions illustrated in B (n = 5–9). *P < 0.05 (Kruskal-Wallis test followed by Dunn <i>post hoc</i> test).</p

Tungstate-mediated activation of BKαβ₁ channels is independent of G proteins.

<p>Representative currents recorded from excised inside-out macropatches obtained from HEK293 cells expressing the BKαβ₁ channels in the presence of 500 μM GDPβS (added to the bath solution) (A) or from transfected HEK293 cells pre-incubated with PTX (500 ng/ml, 24 hours) (C). Currents were recorded at cytosolic 0 Ca²⁺ and 0.7 mM Mg²⁺ before (control, top panels) and 5–10 minutes after cytosolic application of 1 mM tungstate (WO₄^2-, bottom panel). The voltage protocol was as described in the Methods. (B), (D) Average G-V curves for BKαβ₁ channels under the experimental conditions above mentioned. Solid curves were obtained by fitting the normalized conductance to the Boltzmann equation (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0118148#sec002" target="_blank">Methods</a>). (E) Voltage for half maximal activation (V_{1/2 act}) of BKαβ₁ channels before (control, filled circles) and after addition of tungstate (1 mM WO₄^2-, open circles) obtained for the indicated experimental conditions (+GDPβS, n = 4; +PTX, n = 6). No significant difference was found in the decrease on V_{1/2 act} induced by tungstate when comparing both treatments: 24 ± 4 mV (n = 4) for GDPβS treatment <i>versus</i> 16 ± 3 mV (n = 6) for PTX treatment (P = 0.11, Mann-Whitney U-test). Besides the substantial difference in the duration of both treatments to inhibit G proteins (24–28 hours for PTX treatment and minutes for GDPβS treatment), no significant difference was found among them regarding V_{1/2 act} before the addition of tungstate (control situation for GDPβS treatment: V_{1/2 act} = 144 ± 4 mV (n = 4) <i>versus</i> control situation for PTX treatment: V_{1/2 act} = 145 ± 2 mV (n = 6); P > 0.99, Mann-Whitney U-test). Furthermore, these V_{1/2 act} control values (before tungstate application) were similar to the ones previously reported by us under identical experimental conditions but without interfering with the activation of G proteins: 139 ± 2 mV (n = 7) [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0118148#pone.0118148.ref014" target="_blank">14</a>] and 147 ± 2 mV (n = 7) [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0118148#pone.0118148.ref026" target="_blank">26</a>] (P = 0.1, ANOVA). Note that, as previously reported, 1 mM tungstate also reduced substantially the K⁺ current amplitude in the absence of cytosolic Ca²⁺ (A, C), an effect that, contrary to the tungstate-induced reduction of V_{1/2 act}, has been shown to occur either in the absence or presence of Mg²⁺ and in the absence or presence of the different regulatory β subunits (β₁-β₄) [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0118148#pone.0118148.ref014" target="_blank">14</a>]. For each experimental condition V_{1/2 act} and k_act values were (in mV): 144 ± 4 and 20 ± 0.3 (control situation for GDPβS treatment, n = 4); 120 ± 1 and 22 ± 1 (after WO₄^2- addition for GDPβS treatment, n = 4); 145 ± 2 and 19 ± 0.3 (control situation for PTX, n = 6); 129 ± 2 and 21 ± 1 (after WO₄^2- addition for PTX treatment, n = 6). No significant differences were found among k_act values (P = 0.07, ANOVA).</p

BK channels mediate both ERK1/2 phosphorylation and reduction of PDGF-stimulated proliferation induced by tungstate in human vascular smooth muscle cells.

<p>(A) Representative Western-blot showing phosphorylation of ERK1/2 using phospho-ERK-specific antibodies after 10 minutes incubation in control conditions (0% FBS), with PDGF (20 ng/ml), and with tungstate (1 mM) alone or in combination with iberiotoxin (IbTX 100 nM), as indicated. Beta-actin was used as loading control. (B) Protein expression and phosphorylation were quantified by densitometry of the corresponding Western blot signal and normalized to beta-actin. The ratio p-ERK/beta-actin in control conditions was taken as 1, so that fold-changes relative to control are shown. (C) VSMCs were serum starved for 48 hours and then incubated 30 hours with 20 ng/ml PDGF alone or in combination with the indicated compounds. During the last 6 hours of incubation, EdU was added to the media to detect the number of cells entering S-phase. Mean ± S.E.M. of 6–9 determinations from at least four different cultures. ***P < 0.001 (ANOVA followed by Tukey <i>post hoc</i> test).</p

BKαβ₁ channels potentiate tungstate-induced ERK1/2 phosphorylation in a G_i/o protein-dependent manner.

<p>Phosphorylation of ERK1/2 was analyzed by Western blot using phospho-ERK-specific antibodies. Total ERK was used as loading control (data not shown). Protein expression and phosphorylation were quantified by densitometry of the corresponding Western blot signal. Relative density (phosphorylated versus total ERK) was normalized to the inner control (EGF/IGF for each condition, which was considered as 100%). (A) Representative Western blots obtained from HEK293, HEKα and HEKαβ₁ cells for ERK1/2 phosphorylation levels, without treatment (-) or after treatment with 1 mM tungstate (WO₄^2-) (during 5 and 10 minutes, as indicated) in the absence of toxins. (B) Average normalized relative density (phosphorylated versus total ERK) in the absence of toxins. (C) Representative Western blots obtained from HEK293, HEKα and HEKαβ₁ cells for ERK1/2 phosphorylation levels, without treatment (-), after treatment with 100 ng/ml EGF (during 10 minutes) (EGF) or after treatment with 1 mM tungstate (WO₄^2-) (during 5 and 10 minutes, as indicated) in the absence (left) or presence of PTX (right). (D) Average normalized relative density (phosphorylated versus total ERK) in the presence of PTX. (E) Representative Western blots obtained from HEK293, HEKα and HEKαβ₁ cells for ERK1/2 phosphorylation levels, without treatment (-), after treatment with 100 ng/ml IGF (during 10 minutes) (IGF) or after treatment with 1 mM tungstate (WO₄^2-) (during 5 and 10 minutes, as indicated) in the absence (left) or presence of IbTX (right). (F) Average normalized relative density (phosphorylated versus total ERK) in the presence of IbTX. n = 4–12 in each experimental group. *P < 0.05 when compared to the other HEK cell lines (Kruskal-Wallis test followed by Dunn <i>post hoc</i> test). See <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0118148#sec002" target="_blank">Methods</a> for further details.</p