The Role of TRiC-enhanced Actin Folding in Leber Congenital Amaurosis

Purpose: Mutations in TCP-1 ring complex (TRiC) have been associated with Leber Congenital Amaurosis (LCA). TRiC is involved in protein folding and has 8 essential subunits including CCT5. Herein, we studied the retina of TRiC mutant zebrafish to evaluate the possible role of impaired actin and tubulin folding in LCA. Methods: The cct5tf 212b retina was histologically studied using Toluidine Blue staining as well as TUNEL, BrdU-labeling, and Phalloidin assays. Retinal organisation was assessed by quantification of the cellularity utilising DAPI. Results: Laminar organization of cct5tf 212b retinas was intact. Enhanced apoptosis throughout the cct5tf 212b retina was not compensated by higher proliferation rates, leaving the cct5tf 212b retina smaller in size. Quantification of retinal layer cellularity demonstrated that specifically the numbers of the amacrine and the retinal ganglion cells were depleted, suggesting that the cct5tf 212b retina was not uniformly affected by the reduced actin folding. Conclusion: Whereas the current literature suggests that LCA is predominantly affecting retinal photoreceptor cells and the retinal pigment epithelium, cct5tf 212b analyses demonstrated the important role of folding of actin by TRiC, suggesting that cct

Altered microRNA and target gene expression related to Tetralogy of Fallot

MicroRNAs (miRNAs) play an important role in guiding development and maintaining function of the human heart. Dysregulation of miRNAs has been linked to various congenital heart diseases including Tetralogy of Fallot (TOF), which represents the most common cyanotic heart malformation in humans. Several studies have identified dysregulated miRNAs in right ventricular (RV) tissues of TOF patients. In this study, we profiled genome-wide the whole transcriptome and analyzed the relationship of miRNAs and mRNAs of RV tissues of a homogeneous group of 22 non-syndromic TOF patients. Observed profiles were compared to profiles obtained from right and left ventricular tissue of normal hearts. To reduce the commonly observed large list of predicted target genes of dysregulated miRNAs, we applied a stringent target prediction pipeline integrating probabilities for miRNA-mRNA interaction. The final list of disease-related miRNA-mRNA pairs comprises novel as well as known miRNAs including miR-1 and miR-133, which are essential to cardiac development and function by regulating KCNJ2, FBN2, SLC38A3 and TNNI1. Overall, our study provides additional insights into post-transcriptional gene regulation of malformed hearts of TOF patients

Teaching vocabulary to learners with special needs

Diese Diplomarbeit besteht aus einem theoretischen und einem empirischen Teil, welcher eine Fallstudie beinhaltet, die mit einer legasthenischen Lernenden über einen Zeitraum von ungefähr vier Monaten intensiven Lernens durchgeführt wurde. Der theoretische Teil fokussiert das Lehren und Lernen von (englischem)Vokabular im allgemeinen und konzentriert sich auf verschiedene Techniken und Strategien der Präsentation, des Wiederholens und des Testens von Wortschatz. Diese Techniken und Strategien haben zum Ziel, den Lernenden den Zugang zu und das Erinnern von Vokabular zu erleichtern und die Merkfähigkeit zu fördern bzw. zu maximieren. Des weiteren werden im theoretischen Teil, zusätzlich zur Diskussion eines weiten Spektrums von erinnerungsfördernden Techniken, die LehrerInnen in den unterschiedlichen Phasen der Wortschatzarbeit anwenden können, die zugrundeliegenden Theorien und Prinzipien, auf welchen diese Techniken und Strategien gegründet sind sowie die Gründe, warum diese einen essentiellen Part im Lehren und Lernen von Wortschatz darstellen sollten, dargelegt. Darüber hinaus beschäftigt sich der theoretische Teil dieser Arbeit mit der Verwendung einer Vielzahl von Wortschatzwiederholungs- und Wortschatzüberprüfungsaktivitäten und enthält Unterrichtsmaterialien, die im Lehrprojekt verwendet wurden. Der erste Teil des empirischen Teils beschäftigt sich mit drei vom Autor dieser Arbeit aufgestellten Hauptprinzipien effektiver Wortschatzarbeit, gemäß welcher das Lehrprojekt ausgeführt wurde, und liefert außerdem eine kurze Beschreibung der Schwierigkeiten bezüglich des Fremdsprachenerwerbs der Lernenden sowie eine kurze Präsentation der didaktischen Einheiten und des Wortschatzes, mit dem im Unterrichtsprojekt gearbeitet wurde. Der zweite Teil enthält Auszüge und Beispiele des tatsächlichen Unterrichtsprojekts, um die möglichen Auswirkungen der Anwendung der oben genannten Hauptprinzipien auf die Merkfähigkeit der Lernenden zu illustrieren. Bezüglich der dem Unterrichtsprojekt entnommenen Beispiele und Überprüfungen (oder Tests), die dazu dienen die praktische Anwendung der Techniken und deren Auswirkungen darzustellen, muss erwähnt werden, dass der Autor während des Unterrichtsprojektes detaillierte Aufzeichnungen gemacht hat und eine Vielzahl von Tests erstellt hat, um den Lernprozess bzw. den Lernfortschritt der Lernenden zu dokumentieren. Schließlich werden die im Lehrprojekt erhaltenen Daten und Ergebnisse besprochen und reflektiert und mögliche Auswirkungen auf das Lehren und Lernen von Fremdsprachen werden herausgearbeitet.This thesis consists of a theoretical part and an empirical part that includes a case study carried out with a dyslexic learner over a period of approximately four months of intensive study. The theoretical part focuses on vocabulary teaching and learning in general, and on various techniques and strategies of vocabulary presentation, recycling and testing aimed at facilitating and enhancing the learner's accessibility to and retention of lexis. Moreover, besides discussing a wide range of techniques of retention enhancement teachers may apply at the various stages of vocabulary work, the theory part also includes the underlying theories and basic principles which these techniques and strategies are founded on and the reasons why they should form an integral part of vocabulary teaching and learning. Furthermore, the theoretical part also discusses the usage of a large number of vocabulary recycling and testing activities and therefore contains materials used in the teaching project. The first part of the empirical part is concerned with the three main principles of effective vocabulary teaching and learning (proposed by the author of this thesis) according to which the teaching project was carried out and provides a brief description of the learner's difficulties regarding second language acquisition and of the didactic units and the lexis dealt with in the teaching project. The second part includes examples taken from the actual teaching in order to illustrate the impact on the learner's retention of lexis the application of the three main principles might have had. Regarding the examples and tests taken from the teaching project in order to illustrate the actual usage of the techniques and their effects, it has to be mentioned that the author has made use of detailed field notes and designed a variety of tests in order to record the learner's learning process and progress, respectively. Finally, the data and findings will be discussed and reflected on, and possible implications on language teaching and learning will be highlighted

Erfolgsfaktoren beruflich selbstständiger Berufs- und Laufbahnberater

Die qualitative Untersuchung hat zum Ziel, die Erfolgsfaktoren von beruflich selbstständigen Berufs- und Laufbahnberatern zu erfassen. Der theoretische Rahmen umfasst psychologische Untersuchungen, die die Absichtsentstehung, den Zusammenhang mit Laufbahntheorien und die Voraussetzungen für längerfristigen Erfolg erklären. Neben den konkreten Erfolgsfaktoren, zu denen Persönlichkeitsmerkmale, Fähigkeiten und Fertigkeiten und die daraus folgenden Handlungen gehören, werden auch Motive und die persönlichen Definitionen erfragt. Im Zentrum des Interesses stehen diejenigen Faktoren, die auf die handelnde Personen zurückzuführen sind. Sie werden hier als persönliche Erfolgsfaktoren bezeichnete. Um die Fragestellung zu beantworten wurde eine inhaltsanalytische Verfahrensweise angewendet. Mittels eines halbstandardisierten Interviewleitfadens werden 4 Personen befragt. Die Ergebnisse werden einerseits in Form von Falldarstellungen und andererseits in Form eines theoretisch fundierten Ordnungssystems präsentiert. Die Untersuchung zeigt, dass Erfolgsfaktoren aus den Bereichen ökonomische Faktoren, Umweltfaktoren, Berufsbiografie und Persönlichkeitsmerkmale auch bei Berufs- und Laufbahnberatern gefunden werden konnten. Die Ergebnisse bestätigen damit die theoretischen Grundlagen weitgehend. Auf einer niedrigeren Abstraktionsebene konnten spezifische Erfolgsfaktoren identifiziert werden

Norrin stimulates cell proliferation in the superficial retinal vascular plexus and is pivotal for the recruitment of mural cells

Mutations in Norrin, the ligand of a receptor complex consisting of FZD4, LRP5 and TSPAN12, cause severe developmental blood vessel defects in the retina and progressive loss of the vascular system in the inner ear, which lead to congenital blindness and progressive hearing loss, respectively. We now examined molecular pathways involved in developmental retinal angiogenesis in a mouse model for Norrie disease. Comparison of morphometric parameters of the superficial retinal vascular plexus (SRVP), including the number of filopodia, vascular density and number of branch points together with inhibition of Notch signaling by using DAPT, suggest no direct link between Norrin and Notch signaling during formation of the SRVP. We noticed extensive vessel crossing within the SRVP, which might be a loss of Wnt- and MAP kinase-characteristic feature. In addition, endomucin was identified as a marker for central filopodia, which were aligned in a thorn-like fashion at P9 in Norrin knockout (Ndpy/−) mice. We also observed elevated mural cell coverage in the SRVP of Ndpy/− mice and explain it by an altered expression of PDGFβ and its receptor (PDGFRβ). In vivo cell proliferation assays revealed a reduced proliferation rate of isolectin B4-positive cells in the SRVP from Ndpy/− mice at postnatal day 6 and a decreased mitogenic activity of mutant compared with the wild-type Norrin. Our results suggest that the delayed outgrowth of the SRVP and decreased angiogenic sprouting in Ndpy/− mice are direct effects of the reduced proliferation of endothelial cells from the SRV

Novel CRYGC Mutation in Conserved Ultraviolet-Protective Tryptophan (p.Trp131Arg) Is Linked to Autosomal Dominant Congenital Cataract

Congenital cataract (CC), the most prevalent cause of childhood blindness and amblyopia, necessitates prompt and precise genetic diagnosis. The objective of this study is to identify the underlying genetic cause in a Swiss patient with isolated CC. Whole exome sequencing (WES) and copy number variation (CNV) analysis were conducted for variant identification in a patient born with a total binocular CC without a family history of CC. Sanger Sequencing was used to confirm the variant and segregation analysis was used to screen the non-affected parents. The first de novo missense mutation at c.391T>C was identified in exon 3 of CRYGC on chromosome 2 causing the substitution of a highly conserved Tryptophan to an Arginine located at p.Trp131Arg. Previous studies exhibit significant changes in the tertiary structure of the crystallin family in the following variant locus, making CRYGC prone to aggregation aggravated by photodamage resulting in cataract. The variant can be classified as pathogenic according to the American College of Medical Genetics and Genomics (ACMG) criteria (PP3 + PM1 + PM2 + PS2; scoring 10 points). The identification of this novel variant expands the existing knowledge on the range of variants found in the CRYGC gene and contributes to a better comprehension of cataract heterogeneity

Functional Characterization of an In-Frame Deletion in the Basic Domain of the Retinal Transcription Factor ATOH7

Basic helix-loop-helix (bHLH) transcription factors are evolutionarily conserved and structurally similar proteins important in development. The temporospatial expression of atonal bHLH transcription factor 7 (ATOH7) directs the differentiation of retinal ganglion cells and mutations in the human gene lead to vitreoretinal and/or optic nerve abnormalities. Characterization of pathogenic ATOH7 mutations is needed to understand the functions of the conserved bHLH motif. The published ATOH7 in-frame deletion p.(Arg41_Arg48del) removes eight highly conserved amino acids in the basic domain. We functionally characterized the mutant protein by expressing V5-tagged ATOH7 constructs in human embryonic kidney 293T (HEK293T) cells for subsequent protein analyses, including Western blot, cycloheximide chase assays, Förster resonance energy transfer fluorescence lifetime imaging, enzyme-linked immunosorbent assays and dual-luciferase assays. Our results indicate that the in-frame deletion in the basic domain causes mislocalization of the protein, which can be rescued by a putative dimerization partner transcription factor 3 isoform E47 (E47), suggesting synergistic nuclear import. Furthermore, we observed (i) increased proteasomal degradation of the mutant protein, (ii) reduced protein heterodimerization, (iii) decreased DNA-binding and transcriptional activation of a reporter gene, as well as (iv) inhibited E47 activity. Altogether our observations suggest that the DNA-binding basic domain of ATOH7 has additional roles in regulating the nuclear import, dimerization, and protein stability

Selected Rhizosphere Bacteria Help Tomato Plants Cope with Combined Phosphorus and Salt Stresses

Plants are often challenged by multiple abiotic stresses simultaneously. The inoculation of beneficial bacteria is known to enhance plant growth under these stresses, such as phosphorus starvation or salt stress. Here, for the first time, we assessed the efficiency of selected beneficial bacterial strains in improving tomato plant growth to better cope with double stresses in salty and P-deficient soil conditions. Six strains of Arthrobacter and Bacillus with different reservoirs of plant growth-promoting traits were tested in vitro for their abilities to tolerate 2–16% (w/v) NaCl concentrations, and shown to retain their motility and phosphate-solubilizing capacity under salt stress conditions. Whether these selected bacteria promote tomato plant growth under combined P and salt stresses was investigated in greenhouse experiments. Bacterial isolates from Cameroonian soils mobilized P from different phosphate sources in shaking culture under both non-saline and saline conditions. They also enhanced plant growth in P-deficient and salt-affected soils by 47–115%, and their PGP effect was even increased in higher salt stress conditions. The results provide valuable information for prospective production of effective bio-fertilizers based on the combined application of local rock phosphate and halotolerant phosphate-solubilizing bacteria. This constitutes a promising strategy to improve plant growth in P-deficient and salt-affected soils.Peer Reviewe

Novel mutations in CACNA1F and NYX in Dutch families with X-linked congenital stationary night blindness

PURPOSE: To describe the clinical features and genetic analysis of eight X-linked congenital stationary night blindness (XLCSNB) Dutch patients. METHODS: Electroretinogram (ERG) measurements were assessed in Dutch patients. Molecular genetic testing by denaturing high performance liquid chromatography (DHPLC), single stranded conformation polymorphism (SSCP) analysis, and direct sequencing of the CACNA1F and NYX genes were performed in the patients possessing a negative Schubert Bornschein ERG. RESULTS: Molecular genetic testing of CACNA1F and NYX revealed three novel and two known CACNA1F sequence variants as well as two novel sequence alterations in the NYX gene. While one of the CACNA1F sequence variants (5756G>A, R1919H) has been previously described as a common polymorphism in Japanese families, we did not found this transition in 100 European control alleles. CONCLUSIONS: In a pool of eight diagnosed XLCSNB patients, five showed a sequence variation in the CACNA1F and two in the NYX gene. In only one of the eight patients no sequence alteration could be detected. This might be explained by a mutation in other, as yet unidentified coding or regulatory sequences of NYX or CACNA1F or additional genes