MicroRNA-433 Dampens Glucocorticoid Receptor Signaling, Impacting Circadian Rhythm and Osteoblastic Gene Expression

FUNDING This work was supported by the National Institute of Arthritis and Musculoskeletal and Skin Diseases of the National Institutes of Health [AR44877]; the National Institutes for Dental and Craniofacial Research [5T90DE21989]; a Grant-in-Aid award from the American Society for Bone and Mineral Research; the UConn Health Center Research Advisory council; and the Center for Molecular Medicine at UConn Health. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.Peer reviewedPublisher PD

SLUG: a new target of lymphoid enhancer factor-1 in human osteoblasts

<p>Abstract</p> <p>Background</p> <p>Lymphoid Enhancer Factor-1 (Lef-1) is a member of a transcription factor family that acts as downstream mediator of the Wnt/β-catenin signalling pathway which plays a critical role in osteoblast proliferation and differentiation. In a search for Lef-1 responsive genes in human osteoblasts, we focused on the transcriptional regulation of the SLUG, a zinc finger transcription factor belonging to the Snail family of developmental proteins. Although the role of SLUG in epithelial-mesenchymal transition and cell motility during embryogenesis is well documented, the functions of this factor in most normal adult human tissues are largely unknown. In this study we investigated SLUG expression in normal human osteoblasts and their mesenchymal precursors, and its possible correlation with Lef-1 and Wnt/β-catenin signalling.</p> <p>Results</p> <p>The experiments were performed on normal human primary osteoblasts obtained from bone fragments, cultured in osteogenic conditions in presence of Lef-1 expression vector or GSK-3β inhibitor, SB216763. We demonstrated that the transcription factor SLUG is present in osteoblasts as well as in their mesenchymal precursors obtained from Wharton's Jelly of human umbilical cord and induced to osteoblastic differentiation. We found that SLUG is positively correlated with RUNX2 expression and deposition of mineralized matrix, and is regulated by Lef-1 and β-catenin. Consistently, Chromatin Immunoprecipitation (ChIP) assay, used to detect the direct Lef/Tcf factors that are responsible for the promoter activity of SLUG gene, demonstrated that Lef-1, TCF-1 and TCF4 are recruited to the SLUG gene promoter "<it>in vivo</it>".</p> <p>Conclusion</p> <p>These studies provide, for the first time, the evidence that SLUG expression is correlated with osteogenic commitment, and is positively regulated by Lef-1 signal in normal human osteoblasts. These findings will help to further understand the regulation of the human SLUG gene and reveal the biological functions of SLUG in the context of bone tissue.</p

Progressive myoclonus epilepsies due to SEMA6B mutations. New variants and appraisal of published phenotypes

Variants of SEMA6B have been identified in an increasing number of patients, often presenting with progressive myoclonus epilepsy (PME), and to lesser extent developmental encephalopathy, with or without epilepsy. The exon 17 is mainly involved, with truncating mutations causing the production of aberrant proteins with toxic gain of function. Herein, we describe three adjunctive patients carrying de novo truncating SEMA6B variants in this exon (c.1976delC and c.2086C &gt; T novel; c.1978delC previously reported). These subjects presented with PME preceded by developmental delay, motor and cognitive impairment, worsening myoclonus, and epilepsy with polymorphic features, including focal to bilateral seizures in two, and non-convulsive status epilepticus in one. The evidence of developmental delay in these cases suggests their inclusion in the “PME plus developmental delay” nosological group. This work further expands our knowledge of SEMA6B variants causing PMEs. However, the data to date available confirms that phenotypic features do not correlate with the type or location of variants, aspects that need to be further clarified by future studie

A novel de novo HCN1 loss-of-function mutation in genetic generalized epilepsy causing increased neuronal excitability

Abstract The causes of genetic epilepsies are unknown in the majority of patients. HCN ion channels have a widespread expression in neurons and increasing evidence demonstrates their functional involvement in human epilepsies. Among the four known isoforms, HCN1 is the most expressed in the neocortex and hippocampus and de novo HCN1 point mutations have been recently associated with early infantile epileptic encephalopathy. So far, HCN1 mutations have not been reported in patients with idiopathic epilepsy. Using a Next Generation Sequencing approach, we identified the de novo heterozygous p.Leu157Val (c.469C > G) novel mutation in HCN1 in an adult male patient affected by genetic generalized epilepsy (GGE), with normal cognitive development. Electrophysiological analysis in heterologous expression model (CHO cells) and in neurons revealed that L157V is a loss-of-function, dominant negative mutation causing reduced HCN1 contribution to net inward current and responsible for an increased neuronal firing rate and excitability, potentially predisposing to epilepsy. These data represent the first evidence that autosomal dominant missense mutations of HCN1 can also be involved in GGE, without the characteristics of epileptic encephalopathy reported previously. It will be important to include HCN1 screening in patients with GGE, in order to extend the knowledge of the genetic causes of idiopathic epilepsies, thus paving the way for the identification of innovative therapeutic strategies

GWAS meta-analysis of over 29,000 people with epilepsy identifies 26 risk loci and subtype-specific genetic architecture

Epilepsy is a highly heritable disorder affecting over 50 million people worldwide, of which about one-third are resistant to current treatments. Here we report a multi-ancestry genome-wide association study including 29,944 cases, stratified into three broad categories and seven subtypes of epilepsy, and 52,538 controls. We identify 26 genome-wide significant loci, 19 of which are specific to genetic generalized epilepsy (GGE). We implicate 29 likely causal genes underlying these 26 loci. SNP-based heritability analyses show that common variants explain between 39.6% and 90% of genetic risk for GGE and its subtypes. Subtype analysis revealed markedly different genetic architectures between focal and generalized epilepsies. Gene-set analyses of GGE signals implicate synaptic processes in both excitatory and inhibitory neurons in the brain. Prioritized candidate genes overlap with monogenic epilepsy genes and with targets of current antiseizure medications. Finally, we leverage our results to identify alternate drugs with predicted efficacy if repurposed for epilepsy treatment

Genome-wide identification and phenotypic characterization of seizure-associated copy number variations in 741,075 individuals

Copy number variants (CNV) are established risk factors for neurodevelopmental disorders with seizures or epilepsy. With the hypothesis that seizure disorders share genetic risk factors, we pooled CNV data from 10,590 individuals with seizure disorders, 16,109 individuals with clinically validated epilepsy, and 492,324 population controls and identified 25 genome-wide significant loci, 22 of which are novel for seizure disorders, such as deletions at 1p36.33, 1q44, 2p21-p16.3, 3q29, 8p23.3-p23.2, 9p24.3, 10q26.3, 15q11.2, 15q12-q13.1, 16p12.2, 17q21.31, duplications at 2q13, 9q34.3, 16p13.3, 17q12, 19p13.3, 20q13.33, and reciprocal CNVs at 16p11.2, and 22q11.21. Using genetic data from additional 248,751 individuals with 23 neuropsychiatric phenotypes, we explored the pleiotropy of these 25 loci. Finally, in a subset of individuals with epilepsy and detailed clinical data available, we performed phenome-wide association analyses between individual CNVs and clinical annotations categorized through the Human Phenotype Ontology (HPO). For six CNVs, we identified 19 significant associations with specific HPO terms and generated, for all CNVs, phenotype signatures across 17 clinical categories relevant for epileptologists. This is the most comprehensive investigation of CNVs in epilepsy and related seizure disorders, with potential implications for clinical practice

The Role of microRNAs in Osteoclastogenesis

Archivial abstract submitte

Interobserver reproducibility of transabdominal 3-dimensional sonography of the fetal brain.

OBJECTIVE: The purpose of this study was to assess the interobserver reproducibility of transabdominal 3-dimensional (3D) fetal neurosonography. METHODS: This was a prospective observational study. We studied 23 consecutive singleton pregnancies between 18 and 23 weeks' gestation. All cases had normal fetal neurosonographic examination findings, which were confirmed after birth. A 3D sonographic volume of the fetal head was acquired transabdominally by a single operator using an axial approach. Fetal brain anatomy was later analyzed offline by 2 different operators. Axial, sagittal, and coronal views of the fetal brain were obtained to perform a detailed evaluation of the fetal brain. Each operator defined the scanning planes obtained as adequate or inadequate. Results were evaluated with 2 x 2 tables and the Cohen kappa coefficient to assess interobserver agreement. RESULTS: Good-quality multiplanar images were obtained in 23 of 23 cases. The rate of adequate visualization was 100% for all of the axial planes, with kappa values of 1.00. For sagittal and coronal planes, the rate of visualization ranged between 78% and 91%, with kappa values ranging between 0.61 and 0.83. CONCLUSIONS: Transabdominal 3D sonography of the fetal brain at 18 to 23 weeks' gestation has an acceptable degree of interobserver reproducibility

Utilization of Transgenic Models in Evaluation of Osteogenic Differentiation of Embryonic Stem Cells

Previous studies reported that embryonic stem cells (ESCs) can be induced to differentiate into cells showing a mature osteoblastic phenotype by culturing them under osteo-inductive conditions. It is probable that osteogenic differentiation requires that ESCs undergo differentiation through an intermediary step involving a mesenchymal lineage precursor. Based on our previous studies indicating that adult mesenchymal progenitor cells express αSMA, we have generated ESCs from transgenic mice in which an αSMA promoter directs the expression of red fluorescent protein (RFP) to mesenchymal progenitor cells. To track the transition of ESC-derived MSCs into mature osteoblasts, we have utilized a bone-specific fragment of rat type I collagen promoter driving green fluorescent protein (Col2.3GFP). Following osteogenic induction in ESCs, we have observed expression of alkaline phosphatase and subsequent mineralization as detected by von Kossa staining. After one week of osteogenic induction, ESCs begin to express αSMARFP. This expression was localized to the peripheral area encircling a typical ESC colony. Nevertheless, these αSMARFP positive cells did not show activation of the Col2.3GFP promoter, even after 7 weeks of osteogenic differentiation in vitro. In contrast, Col2.3GFP expression was detected in vivo, in mineralized areas following teratoma formation. Our results indicate that detection of alkaline phosphatase activity and mineralization of ESCs cultured under osteogenic conditions is not sufficient to demonstrate osteogenic maturation. Our study indicates the utility of the promoter-visual transgene approach to assess the commitment and differentiation of ESCs into the osteoblast lineage

Linee guida sulla comunicazione on line in tema di tutela e promozione della salute

Questo documento vuole iscriversi in questo processo di rinnovamento del rapporto tra le istituzioni sanitarie e il cittadino con lo scopo di fornire indicazioni (in termini di contenuti informativi, servizi interattivi e strategie comunicative on line) per pianificare e realizzare un’attività di comunicazione in tema di tutela e promozione della salute (anche attraverso un Canale ‘Cittadini’ del portale del Ministero della Salute) che contribuisca a quell’empowerment del cittadino più volte richiamato dall’OMS (1978, 1986, 1998, 2005). L’obiettivo di questo documento è delineare le caratteristiche di un sito di un ente sanitario in grado di offrire una piattaforma telematica capace di incontrare in maniera efficace le esigenze informative, le aspettative e le priorità dei cittadini-pazienti e di favorire in loro lo sviluppo dell’apprendimento di comportamenti di promozione della salute e di prevenzione della malattia, contribuendo anche ad un uso più appropriato dei servizi sanitari. , infatti, la pratica clinica è tanto più efficace quanto più si costruisce su decisioni derivate dall’integrazione tra l’esperienza del medico e l’uso coscienzioso ed esplicito delle migliori evidenze scientifiche disponibili, mediate dalle preferenze del paziente. Partendo da un forte riferimento all’empowerment del cittadino e all’Evidence-based Medicine (EBM), l’epistemologia fondativa del SSN dalla metà degli anni Novanta, questo documento individua due concetti-chiave sui quali fondare la strategia comunicativa dei siti web degli enti e delle strutture sanitarie: la centralità del cittadino-paziente e l’importanza delle informazioni sulle prove di efficacia degli interventi sanitari. Nella prima parte del volume vengono presentati i risultati delle ricerche preparatorie all’individuazione di criteri di qualità per una sito di qualità in materia di tutela e promozione della salute: l’analisi epidemiologica per conoscere la diffusione delle malattie e dei fattori di rischio nella popolazione italiana; un revisione della letteratura relativa ai bisogni informativi di salute dei cittadini stessi, sia in termini di tipologia di informazioni che in termini di percorsi di ricerca preferiti; un’analisi della letteratura scientifica per determinare quale contributo, in termini di efficacia, possa offrire Internet, una rassegna normativa di settore e la ricognizione delle rilevazioni già condotte sul tema della qualità dei siti istituzionali. Vengono poi illustrati i risultati di un’indagine condotta a livello nazionale sull’opinione del cittadino riguardo i bisogni di salute percepiti; e quelli di un’analisi dei portali della salute delle Regioni e delle Aziende Sanitarie Locali che mettono in luce preziose indicazioni che riguardano la caratterizzazione istituzionale dei siti e la relazionalità attivata, il livello di trasparenza amministrativa, la disponibilità dei servizi on line e la qualità tecnologica. Nella seconda parte, invece, vengono presentati ed illustrati le raccomandazioni e i criteri fondamentali per una comunicazione on line di qualità in ambito sanitario, sia in riferimento ai contenuti informativi e alla tipologia degli interventi sanitari in Internet, che in relazione alle strategie di comunicazione e ai criteri redazionali