Inhibition of adenylyl cyclase 8 prevents the upregulation of Orai1 channel, which improves cardiac function after myocardial infarction.

The upregulation of Orai1 and subsequent store-operated Ca2+ entry (SOCE) has been associated with adverse cardiac remodeling and heart failure (HF). However, the mechanism underlying Orai1 upregulation and its role in myocardial infarction remains unclear. Our study investigated the role of Orai1 in activating adenylyl cyclase 8 (AC8) and cyclic AMP (cAMP) response element-binding protein (CREB), as well as its contribution to cardiac dysfunction induced by ischemia and reperfusion (I/R). We found that I/R evoked an increase in the expression of Orai1 and AC8 in rats' hearts, resulting in a substantial rise in diastolic Ca2+ concentration ([Ca2+]i), and reduced ventricular contractions. The expression of Orai1 and AC8 was also increased in ventricular biopsies of post-ischemic HF patients. Mechanistically, we demonstrate that I/R activation of Orai1 stimulated AC8, which produced cAMP and phosphorylated CREB. Subsequently, p-CREB activated the ORAI1 promoter, resulting in Orai1 upregulation and SOCE exacerbation. Intramyocardial administration of AAV9 carrying AC8 short hairpin RNA decreased the expression of AC8, Orai1 and CREB, which restored diastolic [Ca2+]i and improved cardiac contraction. Therefore, our data suggests that the axis composed by Orai1/AC8/CREB plays a critical role in I/R-induced cardiac dysfunction, representing a potential new therapeutic target to limit the progression of the disease toward HF.

The Efficacy and Usability of an Unguided Web-Based Grief Intervention for Adults Who Lost a Loved One During the COVID-19 Pandemic: Randomized Controlled Trial.

BACKGROUND: The death of a loved one was a challenge many people faced during the COVID-19 pandemic within the context of extraordinary circumstances and great uncertainty. Grief is an unavoidable part of life, and for most people, feelings of grief decrease naturally over time. However, for some people, grieving can become a particularly painful process with clinical symptoms that may require professional help to resolve. To provide psychological support to people who had lost a loved one during the COVID-19 pandemic, an unguided web-based psychological intervention was developed. OBJECTIVE: The main objective of this study was to evaluate the efficacy of the web-based treatment, Grief COVID (Duelo COVID in Spanish; ITLAB), in reducing clinical symptoms of complicated grief, depression, posttraumatic stress, hopelessness, anxiety, and suicidal risk in adults. The secondary aim was to validate the usability of the self-applied intervention system. METHODS: We used a randomized controlled trial with an intervention group (IG) and a waitlist control group (CG). The groups were assessed 3 times (before beginning the intervention, upon completing the intervention, and 3 months after the intervention). The intervention was delivered on the web in an asynchronous format through the Duelo COVID web page. Participants created an account that could be used on their computers, smartphones, or tablets. The evaluation process was automated as part of the intervention. RESULTS: A total of 114 participants were randomly assigned to the IG or CG and met criteria for inclusion in the study (n=45, 39.5% completed the intervention and n=69, 60.5% completed the waitlist period). Most participants (103/114, 90.4%) were women. The results indicated that the treatment significantly reduced baseline clinical symptoms in the IG for all variables (P<.001 to P=.006), with larger effect sizes for depression, hopelessness, grief, anxiety, and risk of suicide (all effect sizes ≥0.5). The follow-up evaluation showed that symptom reduction was maintained at 3 months after the intervention. The results from the CG showed that participants experienced significantly decreased levels of hopelessness after completing the time on the waitlist (P<.001), but their suicidal risk scores increased. Regarding the usability of the self-applied intervention system, the results indicated a high level of satisfaction with the Grief COVID. CONCLUSIONS: The self-applied web-based intervention Grief COVID was effective in reducing symptoms of anxiety, depression, hopelessness, risk of suicide risk, posttraumatic stress disorder, and complicated grief disorder. Grief COVID was evaluated by the participants, who reported that the system was easy to use. These results affirm the importance of developing additional web-based psychological tools to help reduce clinical symptoms in people experiencing grief because of the loss of a loved one during a pandemic. TRIAL REGISTRATION: ClinicalTrials.gov NCT04638842; https://clinicaltrials.gov/ct2/show/NCT04638842.

Suicide risk and associated factors in healthcare workers seeking psychological support during COVID-19: a cross-sectional study.

Healthcare workers have been one of the groups most severely affected by the COVID-19 pandemic, leaving them with serious psychological effects. Some of these effects have not been treated promptly, leading to further psychological symptoms. The objective of this study was to evaluate suicide risk in healthcare workers seeking psychological help during the COVID-19 pandemic, and factors associated with this risk on participants that were searching for treatment during the COVID-19 pandemic. This is a cross-sectional study analyzing data from 626 Mexican healthcare workers seeking psychological help due to the COVID-19 pandemic through the www.personalcovid.com platform. Before they entered treatment, the Plutchik Suicide Risk Scale, the Depression Scale of the Center for Epidemiologic Studies, the Pittsburgh Sleep Quality Index, and the Professional Quality of Life Measure, were administered. Results: 49.4% (n = 308) presented suicide risk. The most severely affected groups were nurses (62%, n = 98) and physicians (52.7%, n = 96). Predictors of suicide risk in healthcare workers were secondary traumatic stress, high depressive affect, low positive affect, emotional insecurity and interpersonal problems, and medication use. Conclusions: The suicidal risk detected was high, found mostly in nurses and doctors. This study suggests the presence of psychological effects on healthcare workers, despite the time that has elapsed since the onset of the pandemic.

Efficacy of a transdiagnostic guided internet-delivered intervention for emotional, trauma and stress-related disorders in Mexican population: study protocol for a randomized controlled trial.

BACKGROUND: Emotional and stress-related disorders show high incidence, prevalence, morbidity, and comorbidity rates in Mexico. In recent decades, research findings indicate that cognitive behavioral interventions, from a disorder-specific perspective, are the effective front-line treatment for anxiety and depression care. However, these treatments are not often used. Reasons include limited access and low availability to effective interventions and comorbidity between mental disorders. Emotional deregulation of negative affectivity has been found to be a mediating factor in addressing emotional disorders from a transdiagnostic perspective, aimed at two or more specific disorders. In addition, technological advancement has created alternatives for psychological assistance, highlighting the possibilities offered by technologies since Internet-supported intervention programs have been empirically tested for effectiveness, efficiency and efficacy and can be key to ensuring access to those who are inaccessible. The aim of the study is to evaluate the efficacy, moderators of clinical change and acceptability of a transdiagnostic guided Internet-delivered intervention versus a transdiagnostic self-guided Internet-delivered intervention for emotional, trauma and stress-related disorders, and waiting list in community sample. METHODS: A three-armed, parallel group, superiority randomized controlled clinical trial with repeated measurements at four times: pretest, posttest, follow-up at 3, 6 and 12 months. Outcomes assessor, participant, care provider and investigator will be blinded. Participants aged 18 to 70 years will be randomly allocated 1:1:1 to one of three study arms: a) Transdiagnostic guided internet-delivered intervention with synchronous assistance, b) Transdiagnostic self-guided internet-delivered intervention, c) Waiting list group. Based on sample size estimation, a minimum of 207 participants (69 in each intervention group) will be included. DISCUSSION: The study could contribute to improving the efficacy of transdiagnostic internet-delivered interventions to promote the dissemination of evidence-based treatments and eventually, to decrease the high prevalence of emotional and trauma-related disorders in the Mexican population. TRIAL REGISTRATION: ClinicalTrial.gov: NCT05225701 . Registered February 4, 2022.

Beneficial effects of leptin treatment in a setting of cardiac dysfunction induced by transverse aortic constriction in mouse.

KEY POINTS: Leptin, is a 16 kDa pleiotropic peptide not only primarily secreted by adipocytes, but also produced by other tissues, including the heart. Controversy exists regarding the adverse and beneficial effects of leptin on the heart We analysed the effect of a non-hypertensive dose of leptin on cardiac function, [Ca2+ ]i handling and cellular electrophysiology, which participate in the genesis of pump failure and related arrhythmias, both in control mice and in mice subjected to chronic pressure-overload by transverse aorta constriction. We find that leptin activates mechanisms that contribute to cardiac dysfunction under physiological conditions. However, after the establishment of pressure overload, an increase in leptin levels has protective cardiac effects with respect to rescuing the cellular heart failure phenotype. These beneficial effects of leptin involve restoration of action potential duration via normalization of transient outward potassium current and sarcoplasmic reticulum Ca2+ content via rescue of control sarcoplasmic/endoplasmic reticulum Ca2+ ATPase levels and ryanodine receptor function modulation, leading to normalization of Ca2+ handling parameters. ABSTRACT: Leptin, is a 16 kDa pleiotropic peptide not only primary secreted by adipocytes, but also produced by other tissues, including the heart. Evidence indicates that leptin may have either adverse or beneficial effects on the heart. To obtain further insights, in the present study, we analysed the effect of leptin treatment on cardiac function, [Ca2+ ]i handling and cellular electrophysiology, which participate in the genesis of pump failure and related arrhythmias, both in control mice and in mice subjected to chronic pressure-overload by transverse aorta constriction (TAC). Three weeks after surgery, animals received either leptin (0.36 mg kg-1  day-1 ) or vehicle via osmotic minipumps for 3 weeks. Echocardiographic measurements showed that, although leptin treatment was deleterious on cardiac function in sham, leptin had a cardioprotective effect following TAC. [Ca2+ ]i transient in cardiomyocytes followed similar pattern. Patch clamp experiments showed prolongation of action potential duration (APD) in TAC and leptin-treated sham animals, whereas, following TAC, leptin reduced the APD towards control values. APD variations were associated with decreased transient outward potassium current and Kv4.2 and KChIP2 protein expression. TAC myocytes showed a higher incidence of triggered activities and spontaneous Ca2+ waves. These proarrhythmic manifestations, related to Ca2+ /calmodulin-dependent protein kinase II and ryanodine receptor phosphorylation, were reduced by leptin. The results of the present study demonstrate that, although leptin treatment was deleterious on cardiac function in control animals, leptin had a cardioprotective effect following TAC, normalizing cardiac function and reducing arrhythmogeneity at the cellular level.

Phospholipase A2 as a Molecular Determinant of Store-Operated Calcium Entry.

Activation of phospholipases A2 (PLA2) leads to the generation of biologically active lipid products that can affect numerous cellular events. Ca(2+)-independent PLA2 (iPLA2), also called group VI phospholipase A2, is one of the main types forming the superfamily of PLA2. Beside of its role in phospholipid remodeling, iPLA2 has been involved in intracellular Ca(2+) homeostasis regulation. Several studies proposed iPLA2 as an essential molecular player of store operated Ca(2+) entry (SOCE) in a large number of excitable and non-excitable cells. iPLA2 activation releases lysophosphatidyl products, which were suggested as agonists of store operated calcium channels (SOCC) and other TRP channels. Herein, we will review the important role of iPLA2 on the intracellular Ca(2+) handling focusing on its role in SOCE regulation and its implication in physiological and/or pathological processes.

Proarrhythmic effect of sustained EPAC activation on TRPC3/4 in rat ventricular cardiomyocytes.

The Exchange Protein directly Activated by cAMP (EPAC) participates to the pathological signaling of cardiac hypertrophy and heart failure, in which the role of Ca(2+) entry through the Transient Receptor Potential Canonical (TRPC) channels begin to be appreciated. Here we studied whether EPAC activation could influence the activity and/or expression of TRPC channels in cardiac myocytes. In adult rat ventricular myocytes treated for 4 to 6h with the selective EPAC activator, 8-pCPT (10µM), we observed by Fluo-3 confocal fluorescence a Store-Operated Ca(2+) Entry (SOCE) like-activity, which was blunted by co-incubation with EPAC inhibitors (ESI-05 and CE3F4 at 10 µM). This SOCE-like activity, which was very small in control incubated cells, was sensitive to 30-µM SKF-96365. Molecular screening showed a specific upregulation of TRPC3 and C4 protein isoforms after 8-pCPT treatment. Moreover, sustained EPAC activation favored proarrhythmic Ca(2+) waves, which were reduced either by co-incubation with EPAC inhibitors or bath perfusion with TRPC inhibitors. Our study provides the first evidence that sustained selective EPAC activation leads to an increase in TRPC3 and C4 protein expression and induces a proarrhythmic SOCE-like activity in adult rat ventricular cardiomyocytes, which might be of importance during the development of cardiac diseases.

Internet-based self-administered intervention to reduce anxiety and depression symptomatology and improve well-being in 7 countries: protocol of a randomized control trial.

Background: Online psychological interventions have emerged as a treatment alternative because they are accessible, flexible, personalized, and available to large populations. The number of Internet interventions in Latin America is limited, as are Randomized Controlled Trials (RCTs) of their effectiveness and a few studies comparing their effectiveness in multiple countries at the same time. We have developed an online intervention, Well-being Online, which will be available to the public free of charge in 7 countries: Mexico, Ecuador, Peru, Chile, Brazil, Spain, and the Netherlands. We expect a reduction in depression and anxiety symptoms and an increase in well-being of the participants. Methods: A multi-country, randomized controlled trial will be conducted. The intervention is multicomponent (Cognitive Behavioral Therapy, Behavioral Activation Therapy, Mindfulness, Acceptance and Commitment Therapy, and Positive Psychology), with 10 sessions. In each country, eligible participants will be randomized to one of three groups: Enriched Intervention (interactive web design with videos, infographics, text, audio, and forum), Text Intervention (text on the website), and Wait List (control group). Repeated measures will be obtained at 5-time points. Our primary outcomes will be anxiety symptomatology, depressive symptomatology, and mental well-being. MANOVA analysis will be used for our main analysis. Discussion: This protocol describes the design of a randomized trial to evaluate the efficacy of a web-based intervention to reduce anxiety and depression symptomatology and increase subjective well-being. The intervention will be made available in four languages (Spanish, Portuguese, Dutch, and English). Its results will contribute to the evidence of effectiveness in terms of randomized trials and Internet interventions, mainly in Latin America and Europe.

Epac in cardiac calcium signaling.

Epac, exchange protein directly activated by cAMP, is emerging as a new regulator of cardiac physiopathology. Although its effects are much less known than the classical cAMP effector, PKA, several studies have investigated the cardiac role of Epac, providing evidences that Epac modulates intracellular Ca(2+). In one of the first analyses, it was shown that Epac can increase the frequency of spontaneous Ca(2+) oscillations in cultured rat cardiomyocytes. Later on, in adult cardiomyocytes, it was shown that Epac can induce sarcoplasmic reticulum (SR) Ca(2+) release in a PKA independent manner. The pathway identified involved phospholipase C (PLC) and Ca(2+)/calmodulin kinase II (CaMKII). The latter phosphorylates the ryanodine receptor (RyR), increasing the Ca(2+) spark probability. The RyR, Ca(2+) release channel located in the SR membrane, is a key element in the excitation-contraction coupling. Thus Epac participates in the excitation-contraction coupling. Moreover, by inducing RyR phosphorylation, Epac is arrhythmogenic. A detailed analysis of Ca(2+) mobilization in different microdomains showed that Epac preferently elevated Ca(2+) in the nucleoplasm ([Ca(2+)]n). This effect, besides PLC and CaMKII, required inositol 1,4,5 trisphosphate receptor (IP3R) activation. IP3R is other Ca(2+) release channel located mainly in the perinuclear area in the adult ventricular myocytes, where it has been shown to participate in the excitation-transcription coupling (the process by which Ca(2+) activates transcription). If Epac activation is maintained for some time, the histone deacetylase (HDAC) is translocated out of the nucleus de-repressing the transcription factor myocyte enhancer factor (MEF2). These evidences also pointed to Epac role in activating the excitation-transcription coupling. In fact, it has been shown that Epac induces cardiomyocyte hypertrophy. Epac activation for several hours, even before the cell hypertrophies, induces a profound modulation of the excitation-contraction coupling: increasing the [Ca(2+)]i transient amplitude and cellular contraction. Thus Epac actions are rapid but time and microdomain dependent in the cardiac myocyte. Taken together the results collected indicate that Epac may have an important role in the cardiac response to stress.

Urotensin-II signaling mechanism in rat coronary artery: role of STIM1 and Orai1-dependent store operated calcium influx in vasoconstriction.

OBJECTIVE: Human urotensin-II (UII) is considered the most potentendogenous vasoconstrictor discovered to date, although the precise mechanism activated downstream of its receptor UTS2R in blood vessels remains elusive. The aim of this study was to determine the role of the store operated Ca(2+) entry (SOCE) signaling pathway in UII-induced coronary artery vasoconstriction. METHODS AND RESULTS: We used a combination of isometric tension measurement, Ca(2+) imaging, pharmacology, and molecular approaches to study UII-mediated rat coronary artery vasoconstriction and intracellular Ca(2+) mobilization in coronary smooth muscle cells. We found that UII promoted dose-dependent vasoconstriction and elicited Ca(2+) and Mn(2+) influx, which were sensitive to classical SOCE inhibitors. In addition, knockdown of either STIM1 or Orai1 essentially inhibited UII-mediated SOCE and prevented UII but not high-KCL evoked contraction in transfected coronary artery. Moreover, we found that Ca(2+)-independent phospholipase A(2)ß was involved in UII effects and that is colocalized with STIM1 in different submembrane compartments. Importantly, STIM1 but not Orai1 downregulation inhibits significantly independent phospholipase A(2) activation. Furthermore, lysophosphatidylcholine, an independent phospholipase A(2) product, activated Orai1 but not STIM1-dependent contraction and SOCE. CONCLUSIONS: Here, we demonstrated that different critical players of SOCE signaling pathway are required for UII-induced vasoconstriction of rat coronary artery.

An internet-based self-applied rat phobia treatment using a Virtual Therapy Assistant: Study protocol for a randomized controlled trial.

Specific phobias are a common anxiety disorder that deteriorates the lives of people who suffer from them. To reduce the symptoms produced by this mental disorder exposure therapies have been used. However, low- and middle-income countries, including Mexico, have the lowest rate of treatment due to multiple barriers that prevent addressing mental health problems. Self-applied treatments have been explored in previous studies, nevertheless, high dropout rates are a common problem in this kind of treatment. An alternative is using immersive self-applied treatments that could help increase adherence to the treatment. This article aims to present a study protocol to explore the feasibility of an Internet self-applied exposure treatment for rat phobias, using four types of immersive multimedia elements: images, videos, video games, and 360° videos. Also, the satisfaction and perception of a Virtual Therapy Assistant (VTA) that provides information and support to the user are described. The study protocol will compare two groups of participants, one on the waiting list, and the other will receive the self-applied treatment for rat phobia supported by the VTA. For this study, 45 participants will be recruited and the evaluation measures will be taken at four different times: baseline, post-treatment, and follow-ups at 3 and 6 months. The levels of anxiety and avoidance of the user manifested during the exposure to the multimedia elements, the improvement of the user's clinical symptoms, the level of satisfaction, the perception of effectiveness, and ease of use of the self-applied system will be evaluated. This study is expected to support the viability of self-applied treatment for rat phobias and the use of a VTA, showing the impact on treatment adherence. To the best of our knowledge, this is the first study to explore an exposure treatment for rats using different multimedia elements with the support of a VTA. The work will serve as a basis for the development of new virtual assistants that help in the treatment of other types of specific phobias. This research has been registered in Clinical Trials NCT (NCT05081323).

Measurement scales of mental health related to climate change: a scoping review protocol using artificial intelligence.

INTRODUCTION: Human actions have influenced climate changes around the globe, causing extreme weather phenomena and impacting communities worldwide. Climate change has caused, directly or indirectly, health effects such as injury and physical injuries, which impact morbidity and mortality. Similarly, there is evidence that exposure to climatic catastrophes has serious repercussions on psychological well-being, and rising temperatures and drought have detrimental effects on mental health.Despite the recent effort of researchers to develop specific instruments to assess the effects of climate change on mental health, the evidence on measures of its impact is still scarce, and the constructs are heterogeneous. The aim of this scoping review is to describe the instruments developed and validated to assess the impact of mental health related to climate change. METHODS AND ANALYSIS: This review is registered at Open Science Framework (https://osf.io/zdmbk). This scoping review will follow the reporting elements chosen for systematic review and meta-analysis (Preferred Reporting Items for Systematic Reviews and Meta-Analyses). We proposed a PO question, as it places no restrictions on the participants (P), and the outcome (O) are measurement instruments on mental health related to climate change. A search will be conducted in different databases (PubMed, Scopus, Web of Science, PsycINFO). We will use an open-source artificial intelligence screening tool (ASReview LAB) for the title and abstract review. The full-text review will be performed by three researchers. If there is a disagreement between two independent reviewers, a third reviewer will take the final decision. We will use the COnsensus-based Standards for the selection of health Measurement INstruments tool to assess the risk of bias for each included study. The review will be conducted starting in September 2023. ETHICS AND DISSEMINATION: The planned scoping review does not require ethical approval since it will not involve an ethical risk to the participants. The results obtained from this study will be presented at conferences, congresses and scientific publications.

Online transdiagnostic intervention for emotional, trauma- and stressor-related disorders in the Mexican population: A randomized pilot and feasibility trial.

•Mexican individuals received via telehealth the unified protocol for anxiety, depression, and trauma related disorders.•Online unified protocol reduced depression, anxiety, emotional issues, uncertainty intolerance, and psychological distress.•The transdiagnostic unified protocol is well-suited to train therapists for emotional, trauma, and stressor-related disorders.

Nutritional education knowledge of teachers and nutritionists in four European countries.

Introduction: Objective: the aim of this study was to examine the perception of professionals from four European countries in charge of teaching Nutrition Education (NE) to children in primary schools or hospitals. Methods: this was achieved through an exploratory study that initiated with two focus groups, one with 5 elementary school teachers and another with 14 nutritionists. From the results of it an online survey was designed and distributed internationally to elementary schools and professional clinics in Spain, Italy, Norway, and Austria. The participants were 75 elementary school teachers and 98 nutritionists. It was measured the level of knowledge of teachers and nutritionists to teach NE, and the level of nutritional knowledge of the children in their respective country. Descriptive statistics were conducted, one-factor ANOVAs to analyze the effect of nationality, and when a significant interaction was found, a post-hoc analysis using Bonferroni adjustment was applied. Results: the results indicated that forty-one percent of the participants considered they have "adequate" theoretical knowledge to teach NE. Only 27 % considered they had "adequate" pedagogical training. A significant effect was found: F(3,168) = 17.37, p < 0.001, η2p = 0.24. Regarding the levels of NE knowledge of children, from lowest to highest, there were Spain, Italy, Austria, and Norway. Also, it was observed that professionals and children from Spain and Italy were more affected with less knowledge and training regarding NE. Conclusions: these results could help governments and educational organizations of the affected countries to take decisions to tackle this problematic.

Introducción: Objetivo: el objetivo de este estudio fue examinar la percepción de los profesionales de cuatro países europeos encargados de enseñar educación nutricional (EN) a niños de escuelas primarias u hospitales. Métodos: esto se logró a través de un estudio exploratorio que se inició con dos grupos focales, uno con 5 maestros de primaria y otro con 14 nutricionistas. A partir de los resultados del mismo se diseñó una encuesta en línea y se distribuyó internacionalmente a escuelas primarias y clínicas de profesionales en España, Italia, Noruega, y Austria. Los participantes fueron 75 maestros de primaria y 98 nutricionistas. Se midió el nivel de conocimientos de los maestros y nutricionistas para enseñar EN, y el nivel de conocimientos nutricionales de los niños de su respectivo país. Se hicieron análisis descriptivos, ANOVA de un factor para analizar el efecto de la nacionalidad, y cuando se encontró una interacción significativa se aplicó un análisis post-hoc mediante ajuste de Bonferroni. Resultados: los resultados indicaron que el cuarenta y uno por ciento de los participantes consideró tener conocimientos teóricos "adecuados" para enseñar NE. Solo el 27 % consideró tener una formación pedagógica "adecuada". Se encontró un efecto significativo: F(3,168) = 17.37, p < 0,001, η2p = 0,24. Con respecto a los niveles de conocimiento de NE de los niños en los distintos países, de menor a mayor se clasificaron España, Italia, Austria y Noruega. Asimismo, se observó que los profesionales y los niños de España e Italia se vieron más afectados con menos conocimiento y formación en EN. Conclusiones: estos resultados podrían ayudar a los gobiernos y organizaciones educativas de los países afectados a tomar decisiones para abordar esta problemática.

Sustained Epac activation induces calmodulin dependent positive inotropic effect in adult cardiomyocytes.

Cardiac actions of Epac (exchange protein directly activated by cAMP) are not completely elucidated. Epac induces cardiomyocytes hypertrophy, Ca(2+)/calmodulin protein kinase II (CaMKII) and excitation-transcription coupling in rat cardiac myocytes. Here we aimed to elucidate the pathway cascade involved in Epac sustained actions, as during the initiation of hypertrophy development, where ß-adrenergic signaling is chronically stimulated. Rats were treated with the Epac selective activator 8-pCPT during 4 weeks and Ca(2+) signaling was analyzed in isolated cardiac myocytes by confocal microscopy. We observed a positive inotropic effect manifested by increased [Ca(2+)](i) transient amplitudes. In order to further analyze these actions, we incubated adult cardiomyocytes in the presence of 8-pCPT. The effects were similar to those obtained in-vivo and are blunted by Epac1 knock down. Interestingly, the increase in [Ca(2+)] transients was abolished by protein synthesis blockade or when the downstream effectors of calmodulin (CaMKII or calcineurin) were inhibited, pointing to calmodulin (CaM) as an important downstream protein in Epac sustained actions. In fact, CaM expression was enhanced by 8-pCPT treatment in isolated cells, as found by Western blots. Moreover, the 8-pCPT-induced, PKA-independent, positive inotropic effect was favored by enhanced extracellular Ca(2+) influx via L-type Ca(2+) channels. However, 8-pCPT also induced aberrant Ca(2+) release as Ca(2+) waves and extra [Ca(2+)](i) transients, suggesting proarrhythmic effect. These results provide new insights regarding Epac cardiac actions, suggesting an important role in the initial compensation of the heart to pathological stimuli during the initiation of cardiac hypertrophy, favoring contraction but also arrhythmia risk.

Epac enhances excitation-transcription coupling in cardiac myocytes.

Epac is a guanine nucleotide exchange protein that is directly activated by cAMP, but whose cardiac cellular functions remain unclear. It is important to understand cardiac Epac signaling, because it is activated in parallel to classical cAMP-dependent signaling via protein kinase A. In addition to activating contraction, Ca(2+) is a key cardiac transcription regulator (excitation-transcription coupling). It is unknown how myocyte Ca(2+) signals are decoded in cardiac myocytes to control nuclear transcription. We examine Epac actions on cytosolic ([Ca(2+)](i)) and intranuclear ([Ca(2+)](n)) Ca(2+) homeostasis, focusing on whether Epac alters [Ca(2+)](n) and activates a prohypertrophic program in cardiomyocytes. Adult rat cardiomyocytes, loaded with fluo-3 were viewed by confocal microscopy during electrical field stimulation at 1Hz. Acute Epac activation by 8-pCPT increased Ca(2+) sparks and diastolic [Ca(2+)](i), but decreased systolic [Ca(2+)](i). The effects on diastolic [Ca(2+)](i) and Ca(2+) spark frequency were dependent on phospholipase C (PLC), inositol 1,4,5 triphosphate receptor (IP(3)R) and CaMKII activation. Interestingly, Epac preferentially increased [Ca(2+)](n) during both diastole and systole, correlating with the perinuclear expression pattern of Epac. Moreover, Epac activation induced histone deacetylase 5 (HDAC5) nuclear export, with consequent activation of the prohypertrophic transcription factor MEF2. These data provide the first evidence that the cAMP-binding protein Epac modulates cardiac nuclear Ca(2+) signaling by increasing [Ca(2+)](n) through PLC, IP(3)R and CaMKII activation, and initiates a prohypertrophic program via HDAC5 nuclear export and subsequent activation of the transcription factor MEF2.

A Perspective on How User-Centered Design Could Improve the Impact of Self-Applied Psychological Interventions in Low- or Middle-Income Countries in Latin America.

Global technological progress has generated alternatives for psychological assistance, both for the evaluation and for the treatment of different emotional disorders. Evidence suggests that Internet-based treatments are effective for the treatment of anxiety and depression disorders. However, in Latin America online treatments are still scarce compared to developed countries and have similar problems as developed countries, such as high dropout rate. One possible solution to help decrease the dropout rate is to design and develop online interventions based on the needs and characteristics of the users. The user-centered design (UCD) is a fundamental concept to develop successful online interventions. The objective of this article is to provide a perspective overview on how UCD could improve the impact of self-applied psychological interventions in low- or middle-income countries in Latin America; however this proposal can also be applied in low- and middle-income countries in other regions of the world. The literature on UCD has demonstrated its efficacy when properly applied in online interventions; however, it is not common to see how this methodology has been applied in research in online interventions, and regarding Latin America, this is even more scarce with a very limited number of articles implementing the principles of UCD.

E-Health Psychological Intervention for COVID-19 Healthcare Workers: Protocol for its Implementation and Evaluation.

(1) Background: Healthcare workers have been affected by the COVID-19 pandemic. Digital interventions have been carried out that have been effective with this population; however, few have been reported in Latin America. Our aim is to describe the components and methods to evaluate the feasibility and utility of an online multi-component psychological intervention for healthcare workers in Mexico during COVID-19. (2) Methods: This study is a randomized clinical trial with two arms: (1) self-applied intervention and (2) intervention delivered online by therapists. The participants are randomly assigned to one arm, receiving the same treatment contents in both groups. The "Personal COVID" intervention consists of an internet platform containing 9 nuclear and 3 complementary modules. The objectives of the intervention are: (1) to reduce anxiety, depressive symptoms, burnout, and compassion fatigue, and (2) to increase the quality of life, sleep quality, self-care, and their skills to give bad news. The protocol has been registered on ClinicalTrials.gov (identifier: NCT04890665). (3) Discussion: This protocol is designed according to the highest scientific standards following the SPIRIT guidelines. The "Personal COVID" intervention is expected to be of high efficacy in treating the emotional distress of healthcare workers and promoting their health during the COVID-19 pandemic.

Prevalence of Anxiety Symptoms and Associated Clinical and Sociodemographic Factors in Mexican Adults Seeking Psychological Support for Grief During the COVID-19 Pandemic: A Cross-Sectional Study.

The COVID-19 pandemic is one of the greatest challenges in modern history, with more than four million confirmed deaths worldwide. To date, evidence regarding the psychological impact of the COVID-19 pandemic on grievers is scarce for developing countries such as Mexico. This study aimed to assess the levels of anxiety and associated concerns in a sample of Mexican adults bereaved during the COVID-19 outbreak. A cross-sectional study was conducted through the Duelo COVID (COVID Grief) platform, which is a self-guided online treatment. A total of 5,224 participants reported their anxiety, depression, sleep quality, avoidance, and arousal, prolonged grief symptoms, and medication consumption. Independent sample Mann-Whitney U-tests, chi-square tests, and Kruskal-Wallis tests, as well as multinomial logistic regression, were conducted. Results indicated that 90.4% of the participants reported clinical levels of anxiety, depression, and sleep affectations. The people who lost someone during the last 5 months scored higher in normal grief symptoms compared to the people whose loss was 6 months ago or more, and 9.8% of individuals reported the use of prescription medication, with anxiolytics and antidepressants being the most common. Females, younger respondents, unemployed people with a lower educational level, and participants who disclosed a recent suicide attempt were among those who reported medication consumption. Sleep problems were more frequent in older participants.

Urocortin-2 induces vasorelaxation of coronary arteries isolated from patients with heart failure.

1. Urocortin-2 (Ucn2) is a vasoactive peptide belonging to the corticotrophin-releasing factor (CRF) family that has potent cardiovascular actions. It has been suggested that Ucn2 participates in the pathophysiology of heart failure. However, little is known about the mechanisms underlying the action of Ucn2 in human coronary arteries. The aim of the present study was to assess the effects of Ucn2 on the vascular tone of human coronary arteries dissected from heart failure patients. 2. Human coronary arteries were dissected from the hearts of patients subjected to orthotopic heart transplantation. Coronary arteries were obtained from 17 patients with heart failure due to dilated cardiomyopathy of ischaemic origin in Stage III-IV of the New York Heart Association classification. Changes in tone were measured in arterial rings using force transducers. 3. Application of increasing concentrations of Ucn2 (5-20 nmol/L) to arterial rings precontracted with agonists induced dose-dependent relaxation of the coronary artery, which was independent of endothelial cell activation. Furthermore, the inhibition of the adenylyl cyclase by MDL-12 (100 nmol/L) and protein kinase A (PKA) by H89 (1 µmol/L) prevented Ucn2-mediated relaxation of coronary artery rings. 4. The results of the present study suggest that, in heart failure patients, Ucn2 could be useful in modulating coronary artery circulation independent of endothelial integrity through mechanisms that involve adenylyl cyclase activation and PKA stimulation. The findings warrant further investigation of the role of Ucn2 in circulatory regulation and its potential therapeutic application in heart disease.