Characterization of constitutive and acid-induced outwardly rectifying chloride currents in immortalized mouse distal tubular cells

Thiazides block Na+ reabsorption while enhancing Ca2 + reabsorption in the kidney. As previously demonstrated in immortalized mouse DCT (MDCT) cells, chlorothiazide application induced a robust plasma membrane hyperpolarization, which increased Ca2 + uptake. This essential thiazide-induced hyperpolarization was prevented by the Cl− channel inhibitor 5-Nitro-2-(3-phenylpropylamino) benzoic acid (NPPB), implicating NPPB-sensitive Cl− channels, however the nature of these Cl− channels has been rarely described in the literature. Here we show that MDCT cells express a dominant, outwardly rectifying Cl− current at extracellular pH 7.4. This constitutive Cl− current was more permeable to larger anions (Eisenman sequence I; I− > Br− ≥ Cl−) and was substantially inhibited by > 100 mM [Ca2 +]o, which distinguished it from ClC-K2/Barttin. Moreover, the constitutive Cl− current was blocked by NPPB, along with other Cl− channel inhibitors (DIDS, FFA). Subjecting the MDCT cells to an acidic extracellular solution (pH < 5.5) induced a substantially larger outwardly rectifying NPPB-sensitive Cl− current. This acid-induced Cl− current was also anion permeable (I− > Br− > Cl−), but was distinguished from the constitutive Cl− current by its rectification characteristics, ion sensitivities, and response to FFA. In addition, we have identified similar outwardly rectifying and acid-sensitive currents in immortalized cells from the inner medullary collecting duct (mIMCD-3 cells). Expression of an acid-induced Cl− current would be particularly relevant in the acidic IMCD (pH < 5.5). To our knowledge, the properties of these Cl− currents are unique and provide the mechanisms to account for the Cl− efflux previously speculated to be present in MDCT cells

Aldosterone upregulates transient receptor potential melastatin 7 (TRPM7)

Transient receptor potential melastatin 7 (TRPM7) is a ubiquitously expressed Mg2+-permeable ion channel fused to a C-terminal α-kinase domain. Recently, aldosterone was shown to increase intracellular Mg2+ levels and alter inflammatory signaling in TRPM7-expressing HEK293 cells. This study was undertaken to assess whether these effects were related to an aldosterone-mediated increase of TRPM7 current and/or plasma membrane localization. Using HEK293 cells stably expressing WT-TRPM7, we found that 18-h application of aldosterone significantly increased TRPM7 current and TRPM7 plasma membrane protein expression by 48% and 34%, respectively. The aldosterone-mediated increase of TRPM7 current was inhibited by eplerenone, a mineralocorticoid receptor (MR) blocker, and GSK-650394, an inhibitor of the serum- and glucocorticoid-regulated kinase 1 (SGK1). SGK1 blockade also prevented the aldosterone-induced increase of TRPM7 plasma membrane protein. It was further determined that K1648R-TRPM7, the phosphotransferase-inactive TRPM7 mutant, was unresponsive to aldosterone. Therefore, chronic aldosterone treatment increases the plasma membrane expression of TRPM7, which is associated with an increase of TRPM7 current. This process occurs via an MR-dependent, genomic signaling cascade involving SGK1 and a functioning TRPM7 α-kinase domain. We suggest that this mechanism may be of general relevance when interpreting the effects of aldosterone because the MR receptor is found in multiple tissues, and TRPM7 and SGK1 are ubiquitously expressed

Transient receptor potential melastatin 7 cation channel kinase new player in angiotensin II–induced hypertension

Transient receptor potential melastatin 7 (TRPM7) is a bifunctional protein comprising a magnesium (Mg2+)/cation channel and a kinase domain. We previously demonstrated that vasoactive agents regulate vascular TRPM7. Whether TRPM7 plays a role in the pathophysiology of hypertension and associated cardiovascular dysfunction is unknown. We studied TRPM7 kinase–deficient mice (TRPM7Δkinase; heterozygous for TRPM7 kinase) and wild-type (WT) mice infused with angiotensin II (Ang II; 400 ng/kg per minute, 4 weeks). TRPM7 kinase expression was lower in heart and aorta from TRPM7Δkinase versus WT mice, effects that were further reduced by Ang II infusion. Plasma Mg2+ was lower in TRPM7Δkinase versus WT mice in basal and stimulated conditions. Ang II increased blood pressure in both strains with exaggerated responses in TRPM7Δkinase versus WT groups (P<0.05). Acetylcholine-induced vasorelaxation was reduced in Ang II–infused TRPM7Δkinase mice, an effect associated with Akt and endothelial nitric oxide synthase downregulation. Vascular cell adhesion molecule–1 expression was increased in Ang II–infused TRPM7 kinase–deficient mice. TRPM7 kinase targets, calpain, and annexin-1, were activated by Ang II in WT but not in TRPM7Δkinase mice. Echocardiographic and histopathologic analysis demonstrated cardiac hypertrophy and left ventricular dysfunction in Ang II–treated groups. In TRPM7 kinase–deficient mice, Ang II–induced cardiac functional and structural effects were amplified compared with WT counterparts. Our data demonstrate that in TRPM7Δkinase mice, Ang II–induced hypertension is exaggerated, cardiac remodeling and left ventricular dysfunction are amplified, and endothelial function is impaired. These processes are associated with hypomagnesemia, blunted TRPM7 kinase expression/signaling, endothelial nitric oxide synthase downregulation, and proinflammatory vascular responses. Our findings identify TRPM7 kinase as a novel player in Ang II–induced hypertension and associated vascular and target organ damage

Estimating Heterogeneous Intra-class Correlation Coefficients in Dyadic Ecological Momentary Assessment

A method is described for estimating and testing predictors for influence on the variance of momentary behaviors in dyadic ecological momentary assessment data. Results show that the method allows intraclass correlations of momentary observations from two members of the same couple to vary by observation-level, individual-level and couple-level predictors

Aldosterone signaling through transient receptor potential melastatin 7 cation channel (TRPM7) and its α-kinase domain

We demonstrated a role for the Mg2 + transporter TRPM7, a bifunctional protein with channel and α-kinase domains, in aldosterone signaling. Molecular mechanisms underlying this are elusive. Here we investigated the function of TRPM7 and its α-kinase domain on Mg2 + and pro-inflammatory signaling by aldosterone. Kidney cells (HEK-293) expressing wild-type human TRPM7 (WThTRPM7) or constructs in which the α-kinase domain was deleted (ΔKinase) or rendered inactive with a point mutation in the ATP binding site of the α-kinase domain (K1648R) were studied. Aldosterone rapidly increased [Mg2 +]i and stimulated NADPH oxidase-derived generation of reactive oxygen species (ROS) in WT hTRPM7 and TRPM7 kinase dead mutant cells. Translocation of annexin-1 and calpain-II and spectrin cleavage (calpain target) were increased by aldosterone in WT hTRPM7 cells but not in α-kinase-deficient cells. Aldosterone stimulated phosphorylation of MAP kinases and increased expression of pro-inflammatory mediators ICAM-1, Cox-2 and PAI-1 in Δkinase and K1648R cells, effects that were inhibited by eplerenone (mineralocorticoid receptor (MR) blocker). 2-APB, a TRPM7 channel inhibitor, abrogated aldosterone-induced Mg2 + responses in WT hTRPM7 and mutant cells. In 2-APB-treated ΔKinase and K1648R cells, aldosterone-stimulated inflammatory responses were unchanged. These data indicate that aldosterone stimulates Mg2 + influx and ROS production in a TRPM7-sensitive, kinase-insensitive manner, whereas activation of annexin-1 requires the TRPM7 kinase domain. Moreover TRPM7 α-kinase modulates inflammatory signaling by aldosterone in a TRPM7 channel/Mg2 +-independent manner. Our findings identify novel mechanisms for non-genomic actions of aldosterone involving differential signaling through MR-activated TRPM7 channel and α-kinase

Variability of M giant stars based on Kepler photometry: general characteristics

M giants are among the longest-period pulsating stars which is why their studies were traditionally restricted to analyses of low-precision visual observations, and more recently, accurate ground-based data. Here we present an overview of M giant variability on a wide range of time-scales (hours to years), based on analysis of thirteen quarters of Kepler long-cadence observations (one point per every 29.4 minutes), with a total time-span of over 1000 days. About two-thirds of the sample stars have been selected from the ASAS-North survey of the Kepler field, with the rest supplemented from a randomly chosen M giant control sample. We first describe the correction of the light curves from different quarters, which was found to be essential. We use Fourier analysis to calculate multiple frequencies for all stars in the sample. Over 50 stars show a relatively strong signal with a period equal to the Kepler-year and a characteristic phase dependence across the whole field-of-view. We interpret this as a so far unidentified systematic effect in the Kepler data. We discuss the presence of regular patterns in the distribution of multiple periodicities and amplitudes. In the period-amplitude plane we find that it is possible to distinguish between solar-like oscillations and larger amplitude pulsations which are characteristic for Mira/SR stars. This may indicate the region of the transition between two types of oscillations as we move upward along the giant branch.Comment: 12 pages, 13 figures, accepted for publication in MNRAS. The normalized light curves are available upon reques

Procedures for assessing psychological predictors of injuries in circus artists: a pilot prospective study

Background: Research on psychological risk factors for injury has focused on stable traits. Our objective was to test the feasibility of a prospective longitudinal study designed to examine labile psychological states as risk factors of injury. Methods: We measured psychological traits at baseline (mood, ways of coping and anxiety), and psychological states every day (1-item questions on anxiety, sleep, fatigue, soreness, self-confidence) before performances in Cirque du Soleil artists of the show “O”. Additional questions were added once per week to better assess anxiety (20-item) and mood. Questionnaires were provided in English, French, Russian and Japanese. Injury and exposure data were extracted from electronic records that are kept as part of routine business practices. Results: The 43.9% (36/82) recruitment rate was more than expected. Most artists completed the baseline questionnaires in 15 min, a weekly questionnaire in <2 min and a daily questionnaire in <1 min. We improved the formatting of some questions during the study, and adapted the wording of other questions to improve clarity. There were no dropouts during the entire study, suggesting the questionnaires were appropriate in content and length. Results for sample size calculations depend on the number of artists followed and the minimal important difference in injury rates, but in general, preclude a purely prospective study with daily data collection because of the long follow-up required. However, a prospective nested case-crossover design with data collection bi-weekly and at the time of injury appears feasible. Conclusion: A prospective study collecting psychological state data from subjects who train and work regularly together is feasible, but sample size calculations suggest that the optimal study design would use prospective nested case-crossover methodology

Daily mood, partner support, sexual interest, and sexual activity among adolescent women

This is a post print version of the article. The official published version can be accessed from the link below.Objective: to examine day-to-day associations of coitus, sexual interest, partner emotional support, negative mood and positive mood among adolescent women. Methods: Women (ages 14 – 17 at enrollment; N=146) enrolled from one of three adolescent primary care clinics completed up to five 84-day diaries over a 27-month period. The diaries assessed partner interactions, sexual activity, substance use and mood. Partner-specific measures assessed on each day included partner emotional support (4 items; alpha = 0.94), argument with a partner (no/yes) and coitus (no/yes). Within-day measures assessed marijuana use (no/yes), Positive Mood (3-items; alpha = 0. 86); Negative Mood (3-items; alpha = 0.82) and Sexual Interest (1-item). Lagged measures of mood and sexual activity were included in multivariate models to control for recent mood and sexual behavior effects on current day mood and coitus. Two main analyses were conducted: coitus as a predictor of positive and negative mood; and the role of positive and negative mood as predictors of coitus. Analyses were conducted by multivariate mixed effect regression and mixed effect logistic regression models. Results: Data represent 28,376 days from 146 participants. The average number of diary days was 194 days per participant. Sexual activity was reported on 8.3% of days, with condoms used for 27.0% of these coital events. Marijuana was used on 11% of days. Significant predictors of positive mood on a given day included partner support, marijuana use, and coitus. Negative mood was associated with having an argument with a partner and with prior day coitus. Predictors of coitus on a given day included age (Odds ratio = 1.22), increased coital frequency in previous week (OR = 1.49), coitus on the previous day (1.21), increased same-day sexual interest (OR = 2.8) and decreased same-day negative mood (OR = 0.92). Conclusions: The data demonstrate complex associations of sexual interest, mood, partner interactions and sexual activity

Spectroscopy and modelling of ion charge exchange in energetic discharges used in nuclear fusion

Dissertação de mestrado em Psicologia Clínica da Saúde (Sistémica, Saúde e Família), apresentada à Faculdade de Psicologia e de Ciências da Educação da Universidade de CoimbraA dor crónica representa um problema de saúde pública, que afeta o indivíduo em diversos domínios e, também, a sua família. Assim, o presente estudo delineado por uma metodologia mista (qualitativa/quantitativa) visa identificar as dimensões sobre o impacto da dor crónica no sistema familiar mais destacadas por psicólogos clínicos e investigadores que estudam esta temática. Para atingir este objetivo realizaram-se dois grupos de discussão com a participação de três psicólogos/três investigadores em cada, tendo como base um guião de entrevista semiestruturada. A análise de conteúdo às entrevistas originou uma árvore categorial composta por seis dimensões que parecem refletir as opiniões dos psicólogos/investigadores sobre o impacto que a dor crónica tem no contexto familiar (Subtipologia, Definição, Consequências, Impacto, Intervenção e MFSD). Cada uma destas dimensões compreende várias subdimensões em conformidade com a literatura que concetualiza o tema em estudo. Deste modo, pretende-se promover investigação sobre esta temática e incentivar os profissionais de saúde a intervir junto da família, para além do doente.Chronic pain represents a public health problem, which affects the individual in various ways, including his/her family. This study is based on a mixed method (qualitative/quantitative) and aims to identify the main dimensions of impact of chronic pain on the family system, in the perspective of clinical psychologists and researchers studying this issue. To achieve this goal, two discussion groups, with the participation of three psychologists/three researchers each, were constituted and submitted to semi-structured interviews. The respective content analysis has revealed six dimensions that tend to characterize the impact of chronic pain on the family system (Sub-type, Definition, Treatment, Impact, Intervention and MFSD). Each of these dimensions encompasses sub-dimensions, in accordance with the review of literature. Thus, it is intended to promote research on this topic and encourage health professionals to work with the family, in addition to the patient

Play Patterns of a Group of Nursery School Children

[From the Purpose] The purpose of the present study is threefold: 1. To determine the typical pattern of spontaneous play for a group of twenty-five four year old nursery school children with regard to the relative amounts of time spent alone, with their peers, and with adults. 2. To define the existence and nature of correlations between each of the following: Alone and Peer; Peer and Adult; and Alone and Adult. One would predict, on the basis of previous studies already mentioned, a negative correlation both for Peer and Adult and for Alone and Peer. One would not anticipate a significant correlation between Alone and Adult. Both represent rather immature types of social behavior which would be expected to diminish concurrently as development progresses. 3. To find out whether or not significant differences exist between play patterns in regard to eight variables selected from information contained in the nursery school teachers\u27 yearly reports and in questionnaires submitted to the parents. The eight variables were: a. sex b. number of days per week attending school c. ordinal position d. sex of siblings e. working status of mother f. presence of absence of autoerotic behavior g. success of toilet training h. whether or not the child was characterized as being a worried child. As this part of the study necessitated subdividing the twenty-five children into smaller groups for each variable (according to how they were rated for the particular variable), only very small samples - statistically speaking - could be dealt with. Although the validity of a larger study could not be claimed, it was hope that the results obtained would be suggestive and indicate areas of interest for future research