Reversible cardiac disease features in an inducible CUG repeat RNA-expressing mouse model of myotonic dystrophy.

Myotonic dystrophy type 1 (DM1) is caused by a CTG repeat expansion in the DMPK gene. Expression of pathogenic expanded CUG repeat (CUGexp) RNA causes multisystemic disease by perturbing the functions of RNA-binding proteins, resulting in expression of fetal protein isoforms in adult tissues. Cardiac involvement affects 50% of individuals with DM1 and causes 25% of disease-related deaths. We developed a transgenic mouse model for tetracycline-inducible and heart-specific expression of human DMPK mRNA containing 960 CUG repeats. CUGexp RNA is expressed in atria and ventricles and induced mice exhibit electrophysiological and molecular features of DM1 disease, including cardiac conduction delays, supraventricular arrhythmias, nuclear RNA foci with Muscleblind protein colocalization, and alternative splicing defects. Importantly, these phenotypes were rescued upon loss of CUGexp RNA expression. Transcriptome analysis revealed gene expression and alternative splicing changes in ion transport genes that are associated with inherited cardiac conduction diseases, including a subset of genes involved in calcium handling. Consistent with RNA-Seq results, calcium-handling defects were identified in atrial cardiomyocytes isolated from mice expressing CUGexp RNA. These results identify potential tissue-specific mechanisms contributing to cardiac pathogenesis in DM1 and demonstrate the utility of reversible phenotypes in our model to facilitate development of targeted therapeutic approaches.

A Therapeutic Double Whammy: Transcriptional or Post-transcriptional Suppression of Microsatellite Repeat Toxicity by Cas9.

Microsatellite expansion diseases are caused by unstable tandem repeats of 3-10 nucleotides that become pathogenic beyond a threshold number of copies. Two groups present different approaches to reduce pathogenesis by targeting deactivated Cas9 to either the DNA (Pinto et al., 2017) or the RNA (Batra et al., 2017) repeats with therapeutic potential for several diseases.

A comparative evaluation of clinical and radiographic success rate of pulpotomy in primary molars using antioxidant mix and mineral trioxide aggregate: An in vivo 1-year follow-up study.

INTRODUCTION: Pulpotomy technique is most widely accepted clinical procedure for treating primary teeth with coronal pulp inflammation caused by caries with no involvement of the radicular pulp. AIM AND OBJECTIVE: The aim of the study was to evaluate the success and efficacy of mineral trioxide aggregate (MTA) and antioxidant mix as pulpotomy agents both clinically and radiographically. MATERIALS AND METHODS: A total of forty primary molars in children aged between 6 and 9 years, requiring for pulpotomy procedures, were selected. Random samples distribution was done, antioxidant mix (n = 20) and MTA (n = 20) both were used as pulpotomy agent. Under rubber dam isolation, pulpotomy procedure was performed in all samples followed by restoration with stainless steel crowns. Later, the patients were recalled after 6 and 12 months for clinical and radiographic evaluation. The data were evaluated using Fisher's exact test. RESULTS: Statistically analysis shows no significant difference between the two groups (P > 0.05) with respect to clinical and radiographic success rate, but antioxidant mix showed more efficient result than MTA. CONCLUSION: Antioxidant mix pulpotomy is more biocompatible and cost effective than any other commercially available medicament.

BNANC Gapmers Revert Splicing and Reduce RNA Foci with Low Toxicity in Myotonic Dystrophy Cells.

Myotonic dystrophy type 1 (DM1) is a multisystemic disease caused by an expanded CTG repeat in the 3' UTR of the dystrophia myotonica protein kinase (DMPK) gene. Short, DNA-based antisense oligonucleotides termed gapmers are a promising strategy to degrade toxic CUG expanded repeat (CUGexp) RNA. Nucleoside analogs are incorporated to increase gapmer affinity and stability; however, some analogs also exhibit toxicity. In this study, we demonstrate that the 2',4'-BNANC[NMe] (BNANC) modification is a promising nucleoside analog with high potency similar to 2',4'-LNA (LNA). BNANC gapmers targeting a nonrepetitive region of the DMPK 3' UTR show allele-specific knockdown of CUGexp RNA and revert characteristic DM1 molecular defects including mis-splicing and accumulation of RNA foci. Notably, the BNANC gapmers tested in this study did not induce caspase activation, in contrast to a sequence matched LNA gapmer. This study indicates that BNANC gapmers warrant further study as a promising RNA targeting therapeutic.

Family-Related Factors Associated with Caries Prevalence in the Primary Dentition of 5-6-year-old Children in Urban and Rural Areas of Jabalpur City.

AIM: The purpose of this study was to examine the prevalence of dental caries in primary dentition of 5-6-year-old children in urban and rural areas of Jabalpur city. MATERIALS AND METHODS: The present cross-sectional study was conducted in the rural and urban areas of Jabalpur city, India. A power analysis was carried out to select a representative sample of 5-6-year-old children (n = 408), 204 from government schools and 204 from private schools. Parents were interviewed using a self-structured questionnaire to collect data with regard to variables under evaluation. STATISTICAL ANALYSIS: Collected data were subjected to descriptive analysis using the SPSS 12.0 version. Risk factor association with dental caries was investigated using a stepwise logistic regression analysis with P < 0.05 considered significant. RESULTS: This shows significantly higher decayed missing filled teeth among rural children than urban children. It was seen that 46.5% of children whose mothers were illiterate were affected with dental caries. In urban area, 91.5% of children whereas 77% of children in rural area have parental control on sugar consumption. CONCLUSION: It is important to focus on parents' education level when planning preventive programs for young children. Assessing family-related risk factors is essential when instituting preventive/treatment programs for young children.

Awareness and prevention of patient gag reflex among pedodontists in India: A web-based survey.

AIM: The aim of this study is to devise a reliable and valid web-based survey to predict the awareness level and prevention of patient's gag reflex among Indian pedodontists. MATERIALS AND METHODS: An 11-question predictive gagging survey was created, refined, and tested on 377 pedodontists. The questions focused on age group, common procedure associated with gag reflex and the most common technique adapted by dentists in their clinics to prevent gag. RESULTS: There was no statistically significant difference in gagging reflex among age groups with 53.5% of patients reported anxiety and fear as a main cause of gag; behavioral modification technique was considered as the most reliable method for gagging prevention in 68.5% of patients and there was no statistically significant difference in gagging severity index among patients irrespective of age, causes, and methods used to prevent it. CONCLUSION: The web-based gagging survey established that level of awareness regarding management of patient's gag is significantly low among pedodontists in India and hence is a major hindrance in the clinical practice.

Release of neutrophil extracellular traps by neutrophils stimulated with antiphospholipid antibodies: a newly identified mechanism of thrombosis in the antiphospholipid syndrome.

OBJECTIVE: Antiphospholipid antibodies (aPL), especially those targeting ß2 -glycoprotein I (ß2 GPI), are well known to activate endothelial cells, monocytes, and platelets, with prothrombotic implications. In contrast, the interaction of aPL with neutrophils has not been extensively studied. Neutrophil extracellular traps (NETs) have recently been recognized as an important activator of the coagulation cascade, as well as an integral component of arterial and venous thrombi. This study was undertaken to determine whether aPL activate neutrophils to release NETs, thereby predisposing to the arterial and venous thrombosis inherent in the antiphospholipid syndrome (APS). METHODS: Neutrophils, sera, and plasma were prepared from patients with primary APS (n = 52) or from healthy volunteers and characterized. No patient had concomitant systemic lupus erythematosus. RESULTS: Sera and plasma from patients with primary APS had elevated levels of both cell-free DNA and NETs, as compared to healthy volunteers. Freshly isolated neutrophils from patients with APS were predisposed to high levels of spontaneous NET release. Further, APS patient sera, as well as IgG purified from APS patients, stimulated NET release from control neutrophils. Human aPL monoclonal antibodies, especially those targeting ß2 GPI, also enhanced NET release. The induction of APS NETs was abrogated with inhibitors of reactive oxygen species formation and Toll-like receptor 4 signaling. Highlighting the potential clinical relevance of these findings, APS NETs promoted thrombin generation. CONCLUSION: Our findings indicate that NET release warrants further investigation as a novel therapeutic target in APS.

Do neutrophil extracellular traps contribute to the heightened risk of thrombosis in inflammatory diseases?

Thrombotic events, both arterial and venous, are a major health concern worldwide. Further, autoimmune diseases, such as systemic lupus erythematosus, anti-neutrophil cytoplasmic antibody (ANCA)-associated vasculitis, and antiphospholipid syndrome, predispose to thrombosis, and thereby push the risk for these morbid events even higher. In recent years, neutrophils have been identified as important players in both arterial and venous thrombosis. Specifically, chromatin-based structures called neutrophil extracellular traps (NETs) play a key role in activating the coagulation cascade, recruiting platelets, and serving as scaffolding upon which the thrombus can be assembled. At the same time, neutrophils and NETs are emerging as important mediators of pathogenic inflammation in the aforementioned autoimmune diseases. Here, we first review the general role of NETs in thrombosis. We then posit that exaggerated NET release contributes to the prothrombotic diatheses of systemic lupus erythematosus, ANCA-associated vasculitis, and antiphospholipid syndrome.

Clustering of fMRI data for activation detection using HDR models.

The aim of this work is to perform parametric estimation and detection of activation in fMRI data. The proposed procedure is a segmentation algorithm where clustering is based on estimated parameters for a chosen hemodynamic response (HDR) model. These parameters are estimated for each individual voxel by performing a weighted least-squares nonlinear curve fit to its time series. The parameters are used to decide which voxels are candidates for activation. A segmentation algorithm is executed on a subset of the image voxels, selected based upon fitting parameters. Our procedure will yield activation maps constructed from possibly 2-D (i.e., multi-voxel) regions of activation as opposed to identifying voxels based on individual voxel statistical significance, followed by merging into regions. The approach is intended to reduce false detections, producing "cleaner" activation results without resorting to filtering techniques that may sacrifice spatial resolution.