RBFOX2 is required for establishing RNA regulatory networks essential for heart development.

Human genetic studies identified a strong association between loss of function mutations in RBFOX2 and hypoplastic left heart syndrome (HLHS). There are currently no Rbfox2 mouse models that recapitulate HLHS. Therefore, it is still unknown how RBFOX2 as an RNA binding protein contributes to heart development. To address this, we conditionally deleted Rbfox2 in embryonic mouse hearts and found profound defects in cardiac chamber and yolk sac vasculature formation. Importantly, our Rbfox2 conditional knockout mouse model recapitulated several molecular and phenotypic features of HLHS. To determine the molecular drivers of these cardiac defects, we performed RNA-sequencing in Rbfox2 mutant hearts and identified dysregulated alternative splicing (AS) networks that affect cell adhesion to extracellular matrix (ECM) mediated by Rho GTPases. We identified two Rho GTPase cycling genes as targets of RBFOX2. Modulating AS of these two genes using antisense oligos led to cell cycle and cell-ECM adhesion defects. Consistently, Rbfox2 mutant hearts displayed cell cycle defects and inability to undergo endocardial-mesenchymal transition, processes dependent on cell-ECM adhesion and that are seen in HLHS. Overall, our work not only revealed that loss of Rbfox2 leads to heart development defects resembling HLHS, but also identified RBFOX2-regulated AS networks that influence cell-ECM communication vital for heart development.

Correlation of pacing site in right ventricle with paced QRS complex duration.

BACKGROUND: Pacing from RV mid septum and outflow tract septum has been proposed as a more physiological site of pacing and narrower paced QRS complex duration. The paced QRS morphology and duration in different RV pacing sites is under continued discussion. Hence, this study was designed to address the correlation of pacing sites in right ventricle with paced QRS complex duration. METHODS: Two hundred fifty-two consecutive patients who underwent pacemaker implantation were enrolled. Baseline clinical characteristics were recorded for each patient. All patient underwent fluoroscopy, electrocardiogram and echocardiography post pacemaker implantation. Paced QRS duration was calculated from the leads with maximum QRS duration. RESULTS: Mean paced QRS (pQRS) duration was significantly higher in apical septum group with a mean of 148.9 ±â€¯14.8 m s compared to mid septum (139.6 ±â€¯19.9 m s; p-value 0.003) and RVOT septum (139.6 ±â€¯14.8 m s; p-value 0.002) groups, respectively. There was no significant difference between mid-septal and RVOT septal pQRS duration. On multivariate analysis, female gender, baseline QRS duration and RVOT septal pacing were the only predictors for narrow pQRS duration (<150 msec). CONCLUSION: RV mid-septal and RVOT septal pacing were associated with significantly lower pQRS duration as compared with apical pacing. Based on multivariate analysis RVOT septal pacing appears to be preferred and more physiological pacing site.

Developmentally regulated alternative splicing is perturbed in type 1 diabetic skeletal muscle.

INTRODUCTION: Type 1 diabetic patients can develop skeletal muscle weakness and atrophy by molecular mechanisms that are not well understood. Alternative splicing (AS) is critical for gene expression in the skeletal muscle, and its dysregulation is implicated in muscle weakness and atrophy. Therefore, we investigated whether AS patterns are affected in type 1 diabetic skeletal muscle contributing to skeletal muscle defects. METHODS: AS patterns were determined by reverse transcription-polymerase chain reaction and levels of RNA binding proteins were assessed by Western blot in type 1 diabetic mouse skeletal muscle and during normal mouse skeletal muscle development. RESULTS: Five genes with critical functions in the skeletal muscle are misspliced in type 1 diabetic skeletal muscle, resembling their AS patterns at embryonic stages. AS of these genes undergoes dramatic transitions during skeletal muscle development, correlating with changes in specific RNA binding proteins. CONCLUSION: Embryonic spliced variants are inappropriately expressed in type 1 diabetic skeletal muscle. Muscle Nerve 56: 744-749, 2017.

Reactivation of fetal splicing programs in diabetic hearts is mediated by protein kinase C signaling.

Diabetic cardiomyopathy is one of the complications of diabetes that eventually leads to heart failure and death. Aberrant activation of PKC signaling contributes to diabetic cardiomyopathy by mechanisms that are poorly understood. Previous reports indicate that PKC is implicated in alternative splicing regulation. Therefore, we wanted to test whether PKC activation in diabetic hearts induces alternative splicing abnormalities. Here, using RNA sequencing we identified a set of 22 alternative splicing events that undergo a developmental switch in splicing, and we confirmed that splicing reverts to an embryonic pattern in adult diabetic hearts. This network of genes has important functions in RNA metabolism and in developmental processes such as differentiation. Importantly, PKC isozymes α/ß control alternative splicing of these genes via phosphorylation and up-regulation of the RNA-binding proteins CELF1 and Rbfox2. Using a mutant of CELF1, we show that phosphorylation of CELF1 by PKC is necessary for regulation of splicing events altered in diabetes. In summary, our studies indicate that activation of PKCα/ß in diabetic hearts contributes to the genome-wide splicing changes through phosphorylation and up-regulation of CELF1/Rbfox2 proteins. These findings provide a basis for PKC-mediated cardiac pathogenesis under diabetic conditions.

RNA binding proteins in cardiovascular development and disease.

Congenital heart disease (CHD) is the most common birth defect affecting>1.35 million newborn babies worldwide. CHD can lead to prenatal, neonatal, postnatal lethality or life-long cardiac complications. RNA binding protein (RBP) mutations or variants are emerging as contributors to CHDs. RBPs are wizards of gene regulation and are major contributors to mRNA and protein landscape. However, not much is known about RBPs in the developing heart and their contributions to CHD. In this chapter, we will discuss our current knowledge about specific RBPs implicated in CHDs. We are in an exciting era to study RBPs using the currently available and highly successful RNA-based therapies and methodologies. Understanding how RBPs shape the developing heart will unveil their contributions to CHD. Identifying their target RNAs in the embryonic heart will ultimately lead to RNA-based treatments for congenital heart disease.

A large-scale LC-MS dataset of murine liver proteome from time course of heavy water metabolic labeling.

Metabolic stable isotope labeling with heavy water followed by liquid chromatography coupled with mass spectrometry (LC-MS) is a powerful tool for in vivo protein turnover studies. Several algorithms and tools have been developed to determine the turnover rates of peptides and proteins from time-course stable isotope labeling experiments. The availability of benchmark mass spectrometry data is crucial to compare and validate the effectiveness of newly developed techniques and algorithms. In this work, we report a heavy water-labeled LC-MS dataset from the murine liver for protein turnover rate analysis. The dataset contains eighteen mass spectral data with their corresponding database search results from nine different labeling durations and quantification outputs from d2ome+ software. The dataset also contains eight mass spectral data from two-dimensional fractionation experiments on unlabeled samples.

Quantifying label enrichment from two mass isotopomers increases proteome coverage for in vivo protein turnover using heavy water metabolic labeling.

Heavy water metabolic labeling followed by liquid chromatography coupled with mass spectrometry is a powerful high throughput technique for measuring the turnover rates of individual proteins in vivo. The turnover rate is obtained from the exponential decay modeling of the depletion of the monoisotopic relative isotope abundance. We provide theoretical formulas for the time course dynamics of six mass isotopomers and use the formulas to introduce a method that utilizes partial isotope profiles, only two mass isotopomers, to compute protein turnover rate. The use of partial isotope profiles alleviates the interferences from co-eluting contaminants in complex proteome mixtures and improves the accuracy of the estimation of label enrichment. In five different datasets, the technique consistently doubles the number of peptides with high goodness-of-fit characteristics of the turnover rate model. We also introduce a software tool, d2ome+, which automates the protein turnover estimation from partial isotope profiles.

Role of Psychological Makeup in Psychological Rehabilitation of Acid Attack Victims.

From eve-teasing to more aggressive forms of sexual violence, subjection of women to sexual violence has been on the rise. One heinous form of sexual violence is the acid attack. Acid attack refers to the intentional act of throwing acid on an individual with the intent of harming, torturing, disfiguring, injuring, or killing them. Despite an increase in the number of reported cases of acid attacks, the initial nonavailability of strict legislatures and underreporting of the crime have led to gross underrepresentation of the acid attack victims in the scientific literature. Moreover, most researches focus on the impact of acid attack and not on the process of recovery for these victims. Hence, this paper attempts to explore the role of psychological makeup in the psychological rehabilitation of acid attack victims. This study adopted the homogeneous purposive sampling method. The sample consisted of 30 female victims of acid attack between the age group of 18 to 25 years. The narratives of these victims focusing on their experiences before and after the incidence were collected. Findings of this study indicate that psychological makeup is an important variable that is responsible for the successful recovery from trauma. Nearly all of the victims have shown symptoms of maladaptive psychological makeup after facing a violent event. However, after participation in the rehabilitation program, the victims moved from having maladaptive psychological makeup toward having adaptive psychological makeup. The emergent subthemes comprising maladaptive psychological makeup consists of cognitive distortions, hopelessness, shame, and suicidal ideation; and for adaptive psychological makeup, these are positive life orientation, belief in the just world, and self-efficacy. The insights of the study will contribute to an improved understanding of the recovery process of the acid attack victims and help in planning intervention protocols for them.

The mindfulness trajectories of addressing suicidal behaviour: A systematic review.

BACKGROUND AND OBJECTIVES: Suicidal behaviour has been a persistent concern in medical as well as general settings. Many psychotherapeutic approaches have tried to address suicidal behaviour in different ways. Mindfulness-based interventions (MBIs) have garnered much attention in the last decade because of their treatment efficacy. This systematic review aimed to examine evidence-based research regarding the effectiveness of MBIs as a psychotherapy intervention on suicidality and to deliver suggestions that might help future research. METHOD: The identification of literature was made through an extensive search of the electronic databases, to extract studies relating to the efficacy of MBIs on addressing suicidal behaviour. Additional researches based on library sources were searched manually. The studies' selection was based on a pre-determined inclusion and exclusion criteria as well as the quality of the studies. RESULTS: The present review helped us identify 13 studies, including six randomised controlled trials, two controlled studies and five pre-post observational studies. The findings reported in the studies were mostly favourable to MBIs as an effective intervention strategy for suicidal behaviour. CONCLUSION: MBIs show promising effects as an intervention for suicidal behaviour. However, large scale, high-quality trials with active control, and long term intervention efficacy studies are needed to understand the mechanisms through which MBIs reduce suicidal behaviour.

RBFOX2 is critical for maintaining alternative polyadenylation patterns and mitochondrial health in rat myoblasts.

RBFOX2, which has a well-established role in alternative splicing, is linked to heart diseases. However, it is unclear whether RBFOX2 has other roles in RNA processing that can influence gene expression in muscle cells, contributing to heart disease. Here, we employ both 3'-end and nanopore cDNA sequencing to reveal a previously unrecognized role for RBFOX2 in maintaining alternative polyadenylation (APA) signatures in myoblasts. RBFOX2-mediated APA modulates mRNA levels and/or isoform expression of a collection of genes, including contractile and mitochondrial genes. Depletion of RBFOX2 adversely affects mitochondrial health in myoblasts, correlating with disrupted APA of mitochondrial gene Slc25a4. Mechanistically, RBFOX2 regulation of Slc25a4 APA is mediated through consensus RBFOX2 binding motifs near the distal polyadenylation site, enforcing the use of the proximal polyadenylation site. In sum, our results unveil a role for RBFOX2 in fine-tuning expression of mitochondrial and contractile genes via APA in myoblasts relevant to heart diseases.

Stigma and Discrimination During COVID-19 Pandemic.

The COVID-19 pandemic has been instrumental in creating a dramatic shift from people's need to live in mutual association toward a desire to stigmatize distinctive others. Pandemic seems to be causing othering. Stated simply, stigmatization is a social process set to exclude those who are perceived to be a potential source of disease and may pose threat to the effective social living in the society. Based on the secondary evidence collected from news published online or in print, the present article delves into stigma associated with the COVID-19 pandemic among different social groups in the Indian society and the mounting cases of prejudice based on race, class, and religion. It also presents insights into the varied manifestations, and the deleterious consequences of COVID-19 inspired othering brought to its potential targets in India.

Theoretical Mapping of Suicidal Risk Factors During the COVID-19 Pandemic: A Mini-Review.

Suicide prevention in times of COVID-19 pandemic has become more challenging than ever due to unusual circumstances. The common risk factors identified with regard to suicidal behavior are fear of COVID-19, economic instability, poor access to healthcare facilities, pre-existing psychiatric disorders, and social disconnect. The studies done so far have reported either case studies or have made an effort to understand the risk factors. An understanding of the underlying causal pattern from existing theories, behind these risks, will enable adopting appropriate prevention mechanisms. Hence, this review examines evidence related to risk factors of suicides that occurred during COVID 19 and discusses it in the light of three major theoretical approaches: interpersonal model, stress diathesis model, and cognitive model. The insights obtained from the three viewpoints reveal that perceived burdensomeness, thwarted belongingness, stress sensitivity, cognitive errors such as magnification, catastrophic thinking, arbitrary inference, and mind-reading are likely reasons behind these risk factors for suicide. It is suggested that awareness regarding COVID-19 stressors, use of community-based approaches like gatekeeper training, and brief online psychotherapy by using techniques of mindfulness, interpersonal psychotherapy, and cognitive behavior therapy can be useful in reducing suicide risk during COVID-19.

Effectiveness of mindfulness based cognitive behavior therapy on life satisfaction, and life orientation of adolescents with depression and suicidal ideation.

Suicide and depression are among the most alarming phenomena prevalent throughout the world. Various approaches have tried to explain the intricacies in depression and suicide, as a consequence of faulty psychological adjustment of the individual. Several therapeutic approaches have been developed to strengthen one's coping process, among which cognitive behaviour therapy has shown promising results. Also, mindfulness-based approaches to cognitive behavioural therapy have further accelerated the well-being of such individuals. This study was conducted with an aim to see the effect of mindfulness-based cognitive behaviour therapy on life satisfaction and life orientation in adolescents with depression and suicidal behaviour. A sample of 30 adolescents who scored high on scales of depression and suicidal tendencies were administered pre-test measures on life satisfaction and life orientation. After that they were exposed to an eight weeks programme on mindfulness-based cognitive behaviour therapy, followed by a post-assessment on the same measures. The analysis of pre and post test revealed a significant enhancement in life satisfaction, life orientation, and family functioning as well as a reduction in depressive symptoms and suicidal ideation. It is concluded that mindfulness-based cognitive behaviour therapy serves as an effective medium to enhance the psychological functioning of depressive and suicidal adolescents.

Prevalence of hypertension among Indian adults: Results from the great India blood pressure survey.

OBJECTIVE: Hypertension is the most important risk factor for cardiovascular morbidity and mortality. There is limited data on hypertension prevalence in India. This study was conducted to estimate the prevalence of hypertension among Indian adults. METHODS: A national level survey was conducted with fixed one-day blood pressure measurement camps across 24 states and union territories of India. Hypertension was defined as systolic blood pressure (BP) ≥140 mmHg or a diastolic BP ≥90 mmHg or on treatment for hypertension. The prevalence was age- and gender-standardized according to the 2011 census population of India. RESULTS: Blood pressure was recorded for 180,335 participants (33.2% women; mean age 40.6 ± 14.9 years). Among them, 8,898 (4.9%), 99,791 (55.3%), 35,694 (11.9%), 23,084 (12.8%), 9,989 (5.5%), and 2,878 (1.6%) participants were of the age group 18-19, 20-44, 45-54, 55-64, 65-74, and ≥ 75 years, respectively. Overall prevalence of hypertension was 30.7% (95% confidence interval [CI]: 30.5, 30.9) and the prevalence among women was 23.7% (95% CI: 23.3, 24). Prevalence adjusted for 2011 census population and the WHO reference population was 29.7% and 32.8%, respectively. CONCLUSION: There is a high prevalence of hypertension, with almost one in every three Indian adult affected.

The tumour suppressor RASSF1A is a novel substrate of PKC.

Ras association domain family 1A (RASSF1A) is a tumour suppressor that contains an amino-terminal cysteine-rich region, similar to the diacylglycerol (DAG)-binding domain (C1 domain) found in the protein kinase C (PKC) family of proteins, and a carboxy-terminal Ras-association (RA) domain. In the present study, RASSF1A was identified as a substrate for PKC. Using classical biochemical approaches, it was established that S197 and S203 within the RA domain of RASSF1A are phosphorylated by PKC in vitro and in vivo. Unlike the WT protein, the S197, 203D double mutant of RASSF1A failed to modulate microtubule organization and perinuclear vimentin collapse. By contrast, the equivalent AA mutant of RASSF1A phenocopied the WT protein. These findings indicate that PKC phosphorylation of RASSF1A regulates its ability to reorganize the microtubule network.

Fortilin potentiates the peroxidase activity of Peroxiredoxin-1 and protects against alcohol-induced liver damage in mice.

Fortilin, a pro-survival molecule, inhibits p53-induced apoptosis by binding to the sequence-specific DNA-binding domain of the tumor suppressor protein and preventing it from transcriptionally activating Bax. Intriguingly, fortilin protects cells against ROS-induced cell death, independent of p53. The signaling pathway through which fortilin protects cells against ROS-induced cell death, however, is unknown. Here we report that fortilin physically interacts with the antioxidant enzyme peroxiredoxin-1 (PRX1), protects it from proteasome-mediated degradation, and keeps it enzymatically active by blocking its deactivating phosphorylation by Mst1, a serine/threonine kinase. At the whole animal level, the liver-specific overexpression of fortilin reduced PRX1 phosphorylation in the liver, enhanced PRX1 activity, and protected the transgenic animals against alcohol-induced, ROS-mediated, liver damage. These data suggest the presence of a novel oxidative-stress-handling pathway where the anti-p53 molecule fortilin augments the peroxidase PRX1 by protecting it against degradation and inactivation of the enzyme. Fortilin-PRX1 interaction in the liver could be clinically exploited further to prevent acute alcohol-induced liver damage in humans.

Rbfox2 function in RNA metabolism is impaired in hypoplastic left heart syndrome patient hearts.

Hypoplastic left heart syndrome (HLHS) is a fatal congenital heart disease in which the left side of the heart is underdeveloped, impairing the systemic circulation. Underdeveloped left ventricle exerts biomechanical stress on the right ventricle that can progress into heart failure. Genome-wide transcriptome changes have been identified at early stages in the right ventricle (RV) of infants with HLHS, although the molecular mechanisms remain unknown. Here, we demonstrate that the RNA binding protein Rbfox2, which is mutated in HLHS patients, is a contributor to transcriptome changes in HLHS patient RVs. Our results indicate that majority of transcripts differentially expressed in HLHS patient hearts have validated Rbfox2 binding sites. We show that Rbfox2 regulates mRNA levels of targets with 3'UTR binding sites contributing to aberrant gene expression in HLHS patients. Strikingly, the Rbfox2 nonsense mutation identified in HLHS patients truncates the protein, impairs its subcellular distribution and adversely affects its function in RNA metabolism. Overall, our findings uncover a novel role for Rbfox2 in controlling transcriptome in HLHS.

Dysregulation of RBFOX2 Is an Early Event in Cardiac Pathogenesis of Diabetes.

Alternative splicing (AS) defects that adversely affect gene expression and function have been identified in diabetic hearts; however, the mechanisms responsible are largely unknown. Here, we show that the RNA-binding protein RBFOX2 contributes to transcriptome changes under diabetic conditions. RBFOX2 controls AS of genes with important roles in heart function relevant to diabetic cardiomyopathy. RBFOX2 protein levels are elevated in diabetic hearts despite low RBFOX2 AS activity. A dominant-negative (DN) isoform of RBFOX2 that blocks RBFOX2-mediated AS is generated in diabetic hearts. DN RBFOX2 interacts with wild-type (WT) RBFOX2, and ectopic expression of DN RBFOX2 inhibits AS of RBFOX2 targets. Notably, DN RBFOX2 expression is specific to diabetes and occurs at early stages before cardiomyopathy symptoms appear. Importantly, DN RBFOX2 expression impairs intracellular calcium release in cardiomyocytes. Our results demonstrate that RBFOX2 dysregulation by DN RBFOX2 is an early pathogenic event in diabetic hearts.

Decalepisarayalpathra (J. Joseph & V. Chandras.) Venter, an endemic and endangered ethno medicinal plant from Western Ghats, India.

Decalepis arayalpatra is an endemic and critically endangered plant of India. May 2014 issue of Natural Products Research publishes the findings of R. S. Verma et al. on the chemical composition of D. arayalpatra. This study was conducted to characterise the root aroma of this plant for possible industrial applications. The authors suggest that due to its peculiar vanilla flavour, the plant could be explored as a potential substitute of vanillin-aroma in the flavour industry. Owing to the fact that D. arayalpatra is a critically endangered plant species, and its habitat is now limited to only the protected areas and reserve forest in southern part of India, and that collecting any plant from such reserve forests for commercial activities is illegal as per the law of the country, this specific conclusion of the authors is totally un-substantiated by the law of land, hence, calls for further review.