Functional characterization of the synaptic-activity regulated gene Synaptotagmin10

During epileptogenesis multiple cellular and molecular changes occur in the brain resulting in the emergence of recurrent spontaneous seizures. One member of the Synaptotagmin (Syt) gene family that is rapidly and transiently up-regulated in the rat hippocampus early after the experimental induction of SE but is barely detectable in the native brain is Syt10. However, the knowledge about the function of Syt10 in health and disease conditions, especially in the hippocampus, is very limited. In order to delineate the mechanisms regulating Syt10 gene expression in response to strong electrical stimulation, this study provided the first fundamental bioinformatic and functional characterization of the Syt10 promoter and genomic regulatory regions in different cell types. Conserved regions in the Syt10 gene mediated strong induction of Syt10 promoter activity, particularly in the hippocampal formation. The transcription factor NPAS4 was identified to positively regulate endogenous expression of Syt10 as well as, together with Per1, Syt10 promoter activity. Epilepsy-induced up-regulation of endogenous Syt10 expression was mimicked and a key genomic region underlying depolarization-mediated induction of the Syt10 promoter was clearly identified. As a way to obtain first insights into the putative function of Syt10, its subcellular distribution was examined in hippocampal neurons and was compared with the related Syt isoforms 3, 5 and 6. Syt10 was identified not to be a resident Golgi protein but partially co-localized with IGF1 and with NPY. These results support the hypothesis that Syt10 is localized to large vesicular structures but not to synaptic vesicles. Furthermore, targeting signals defining the subcellular localization of Syt10 were analyzed. Additionally, first steps have been accomplished regarding the identification of Syt10 containing protein complexes or organelles forming the basis for an isolation of the respective complexes from the hippocampus following status epilepticus (SE). To date, the role of Syt10 in epileptogenesis remains enigmatic and the spatiotemporal expression profile was only analyzed in rats. Therefore, the time course of Syt10 expression following pilocarpine-induced SE was examined in mice. The strongest induction of Syt10 expression in mice was observed with a 30 hours delay compared to rats but did not show an exact parallelism to the IGF1 augmentation. However, as reported for IGF1, Syt10 was observed to be important for SE-induced progenitor cell proliferation as well. These data may suggest that Syt10 plays a critical role in the attenuation of acquired changes during epileptogenesis. Taken together, the results of this thesis provide novel insights into the key mechanisms regulating Syt10 expression in the hippocampus in the normal and epileptic brain as well as into the subcellular distribution of Syt10 and form the basis for further studies into the precise understanding of the function of Syt10 during epileptogenesis

Modulation of chloride homeostasis by inflammatory mediators in dorsal root ganglion neurons

<p>Abstract</p> <p>Background</p> <p>Chloride currents in peripheral nociceptive neurons have been implicated in the generation of afferent nociceptive signals, as Cl^-accumulation in sensory endings establishes the driving force for depolarizing, and even excitatory, Cl^-currents. The intracellular Cl^-concentration can, however, vary considerably between individual DRG neurons. This raises the question, whether the contribution of Cl^-currents to signal generation differs between individual afferent neurons, and whether the specific Cl^-levels in these neurons are subject to modulation. Based on the hypothesis that modulation of the peripheral Cl^-homeostasis is involved in the generation of inflammatory hyperalgesia, we examined the effects of inflammatory mediators on intracellular Cl^-concentrations and on the expression levels of Cl^-transporters in rat DRG neurons.</p> <p>Results</p> <p>We developed an <it>in vitro </it>assay for testing how inflammatory mediators influence Cl^-concentration and the expression of Cl^-transporters. Intact DRGs were treated with 100 ng/ml NGF, 1.8 μM ATP, 0.9 μM bradykinin, and 1.4 μM PGE₂for 1–3 hours. Two-photon fluorescence lifetime imaging with the Cl^--sensitive dye MQAE revealed an increase of the intracellular Cl^-concentration within 2 hours of treatment. This effect coincided with enhanced phosphorylation of the Na⁺-K⁺-2Cl^-cotransporter NKCC1, suggesting that an increased activity of that transporter caused the early rise of intracellular Cl^-levels. Immunohistochemistry of NKCC1 and KCC2, the main neuronal Cl^-importer and exporter, respectively, exposed an inverse regulation by the inflammatory mediators. While the NKCC1 immunosignal increased, that of KCC2 declined after 3 hours of treatment. In contrast, the mRNA levels of the two transporters did not change markedly during this time. These data demonstrate a fundamental transition in Cl^-homeostasis toward a state of augmented Cl^-accumulation, which is induced by a 1–3 hour treatment with inflammatory mediators.</p> <p>Conclusion</p> <p>Our findings indicate that inflammatory mediators impact on Cl^-homeostasis in DRG neurons. Inflammatory mediators raise intracellular Cl^-levels and, hence, the driving force for depolarizing Cl^-efflux. These findings corroborate current concepts for the role of Cl^-regulation in the generation of inflammatory hyperalgesia and allodynia. As the intracellular Cl^-concentration rises in DRG neurons, afferent signals can be boosted by excitatory Cl^-currents in the presynaptic terminals. Moreover, excitatory Cl^-currents in peripheral sensory endings may also contribute to the generation or modulation of afferent signals, especially in inflamed tissue.</p

Habit reversal therapy in obsessive compulsive related disorders : A systematic review of the evidence and consort evaluation of randomized controlled trials

Background: Habit Reversal Therapy (HRT) has long been used in the treatment of Tourette Syndrome and Tic Disorders. It has more recently been used to treat Trichotillomania and skin picking behaviors, both considered as Obsessive Compulsive Related Disorders (OCRD). Objectives: This literature review sought to establish and quality assess the existing randomized controlled trial evidence supporting the use of HRT in the DSM-5 family of OCRDs. Search Methods: EMBASE, PsycINFO, PubMed, and Cochrane databases were searched for key terms relating to each OCRD (as classified in the DSM-5), and HRT. Selection Criteria: Titles and abstracts were screened, and any literature matching pre-specified criteria were then selected to be reviewed further. Of these, 8 Randomized Controlled Trials (RCT) relating to Trichotillomania, and 2 RCTs relating to Excoriation Disorder, were extracted and reviewed against the 2010 Consolidating Standards of Reporting Trials (CONSORT) statement. Results: The review identified 10 RCTs of HRT, but these were limited to patients with a primary diagnosis of Trichotillomania or “excoriation behavior,” only. There were some reports of the use of HRT in Tourette Syndrome or Tic Disorder with secondary OCD, but the OCD symptoms were not reliably reported on. Conclusion: There is a gap in the current literature regarding the use of HRT in the DSM-5 OCRDs. In those RCTs that have been reported, the quality of study methodology was questionable as evaluated by CONSORT criteria. The implications of these findings are discussed, and suggestions are made for future research.Peer reviewe

Implementation and Evaluation of Therapeutic Online Coaching Using Habit Reversal Training in Children With Tourette's Disorder - A Pilot Study

Cognitive-behavioral interventions can be difficult to implement in daily routine, which is often essential for generalizing treatment effects to natural settings. Furthermore, there is a lack of adequate care options concerning habit reversal training for children with Tourette's disorder. The objective of this study is to evaluate therapeutic online coaching via videoconferencing in the natural environment of children with Tourette's disorder in addition to face-to-face therapy (blended therapy). Online coaching took place twice a week for a maximum of 12 weeks. In a single-case study (n = 5; patients aged 8-11 years), the first results were obtained for exploratory purposes, especially with regard to the feasibility and reduction of symptoms and impairment. Various outcome measures were assessed (severity of symptoms, impairment, practical implementation, and satisfaction). Despite some principal limitations, the findings provide first hints that blended therapy is feasible and improves symptoms in some children with tics

Webcam-Based Online Coaching with Children and Adolescents with Obsessive-Compulsive Disorders – A Single-Case Study

Objective: Although there is clear evidence-based knowledge regarding state-of-the-art treatment for pediatric obsessive-compulsive disorder (OCD), two main issues remain in clinical practice: (1) Exposure-based cognitive behavioral therapy (CBT) is limited in terms of availability and accessibility or is not adequately provided, and (2) despite large effect sizes of exposure-based CBT, the achieved recovery rates of 50-60 % still show room for improvement. These issues have prompted an increasing focus on delivering exposure-based CBT in new and innovative ways. This study aims to evaluate an intensive therapist-administered online coaching program consisting of exposure with response prevention via video teleconferencing (VTC) as an add-on to weekly outpatient CBT (blended therapy). Method: The blended therapy is examined in n = 5 children and adolescents with OCD using an AB design and multi-informant ratings. Results: This single-case study shows promising results, indicating that a decrease in OCD severity and related functional impairment can be attributed to blended therapy. Moreover, satisfaction with online coaching was high. Conclusions: Despite some principal limitations, the results support the effectiveness and feasibility of blended therapy

Webcam-Based Online Coaching with Children and Adolescents with Obsessive-Compulsive Disorders - A Single-Case Study

Objective: Although there is clear evidence-based knowledge regarding state-of-the-art treatment for pediatric obsessive-compulsive disorder (OCD), two main issues remain in clinical practice: (1) Exposure-based cognitive behavioral therapy (CBT) is limited in terms of availability and accessibility or is not adequately provided, and (2) despite large effect sizes of exposure-based CBT, the achieved recovery rates of 50-60% still show room for improvement. These issues have prompted an increasing focus on delivering exposure-based CBT in new and innovative ways. This study aims to evaluate an intensive therapist-administered online coaching program consisting of exposure with response prevention via video teleconferencing (VTC) as an add-on to weekly outpatient CBT (blended therapy). Method: The blended therapy is examined inn = 5 children and adolescents with OCD using an AB design and multi-informant ratings. Results: This single-case study shows promising results, indicating that a decrease in OCD severity and related functional impairment can be attributed to blended therapy. Moreover, satisfaction with online coaching was high. Conclusions: Despite some principal limitations, the results support the effectiveness and feasibility of blended therapy

Assessment of Self-Concept of Children and Adolescents. Validation of a Modified Version of Harter's Self-Perception Profile for Children in a Clinical Sample

Theoretical Background: The Self-Perception Profile for Children (abbr. SPPC) is widely used internationally and assesses the self-concept of children and adolescents. Purpose: The psychometric properties of a modified version of the SPPC will be tested for a German clinical sample (n = 195, age range 6- 18 years). Methods: Confirmatory factor analyzes, reliability analyzes and analyzes for convergent, divergent and differential validity are considered. Results: The bifactor model and the model with five correlated factors have the best fit indices. The internal consistencies of the scales are satisfactory. The omega statistics indicate psychometric weaknesses. Evidence for convergent, divergent and differential validity can be found. Conclusions: The modified version of the SPPC can be used to assess the self-concept of children and adolescents for a clinical sample; adaptations to optimize the quality of the instrument are discussed

Telephone-assisted self-help for parents of children with attention-deficit/hyperactivity disorder who have residual functional impairment despite methylphenidate treatment: a randomized controlled trial

BackgroundSelf-help parenting interventions have been shown to be effective in the management of children with attention-deficit/hyperactivity disorder (ADHD) and may be useful when there are barriers to face-to-face therapist-led parent trainings. Previous studies indicate that behavioral interventions might be a useful adjunct to medication in children with residual ADHD symptoms, and regarding comorbid oppositional symptoms and multiple domains of functional impairment. In the present study, we examined whether a telephone-assisted self-help (TASH) parenting behavioral intervention (written materials plus telephone counseling) enhanced the effects of methylphenidate treatment in children with ADHD. MethodsIn this randomized controlled trial, parents of 103 school-aged children with ADHD and residual functional impairment despite methylphenidate treatment were randomly assigned to either the enhancement group, which received the TASH intervention as adjunct to routine clinical care (including continued medication), or to the active control group, which received routine clinical care only (including continued medication). Parent-completed outcome measures at baseline and at 12months (postassessment) included functional impairment, ADHD symptoms, oppositional defiant disorder (ODD) symptoms, parenting behavior, and parental satisfaction with the intervention (ClinicalTrials.gov: NCT01660425; URL: ). ResultsIntention-to-treat analyses of covariance (ANCOVAs), which controlled for baseline data, revealed significant and moderate intervention effects for ODD symptoms and negative parenting behavior at the postassessment, whereas per-protocol analyses additionally showed significant and moderate effects on functional impairment (primary outcome). Parents expressed high satisfaction with the program. ConclusionsThe TASH program enhances effects of methylphenidate treatment in families who complete the intervention. Thediscontinuation rate of about 30% and comparison between completing and discontinuing families suggest that the program may be more suitable for families with a higher educational level and fewer additional stresses

A new treatment for children with chronic tic disorders - Resource activation

The aim of this pilot study is to evaluate the efficacy of a resource activation (RA) program as an alternative intervention for children and adolescents with tic disorders. RA interventions focus on the strengths and abilities of the patients. This is expected to improve the patients' perceptions regarding their own resources, which may indirectly result in a reduction of tic symptoms, impairment and comorbid conditions. A within-subject design with two phases (8 weeks diagnostic; 16 sessions treatment) was analyzed using multilevel modeling (n = 24). During the treatment phase, significant reductions of tics were found in clinical rating (Yale Global Tic Severity Scale; YGTSS), parent rating and video observation (motor tics). Moreover, an improvement was shown on most tic-related impairment and subjective burden (SB) ratings. No significant improvement was found regarding comorbid problems and self-esteem. Compared to the preceding diagnostic phase, a significant incremental treatment effect emerged in clinical rating of tic symptoms (YGTSS) and video observation (motor tics). This pilot study provides first hints that RA may represent an effective treatment for reducing tic symptoms, impairment and SB. However, further research is needed in order to establish RA as an effective treatment for tic disorders