Bilder in Texten – Analyse von Eingewöhnungskonzepten mit der dokumentarischen Methode

Der Beitrag fokussiert das methodische Vorgehen bei der Analyse von Eingewöhnungskonzepten als Konzeptpapieren in Schrift und Bild, wie sie von Kindertageseinrichtungen selbst erstellt bzw. an Eltern und weitere Adressat_innen ausgegeben werden. Rahmen der Analyse bildet das Forschungsprojekt „Eingewöhnungskonzepte zwischen Programmatik und Praxis“, in dem untersucht wird, wie pädagogische Fachkräfte in und mit solchen Konzepten ihren gesellschaftlichen Auftrag zur Betreuung der Kinder interpretieren und wie sie öffentliche und familiale Kleinkindbetreuung dabei ins Verhältnis setzen. Die Beispielanalyse konzentriert sich hier vornehmlich auf das Bildmaterial in solchen Dokumenten, die von Einrichtungen im Internet publiziert wurden. Mit der Dokumentarischen Methode – die als etablierte Methode bislang vor allem zur Analyse von Interviews und Gruppendiskussionen, aber auch Bilddokumenten eingesetzt wird – wird dafür ein Analyseverfahren der rekonstruktiven Sozialforschung genutzt, um empirischen Zugang zur konzeptionellen Ebene frühpädagogischer Praxis zu gewinnen. Durch die schrittweise nachvollziehbare formulierende und reflektierende Interpretation einer Abbildung auf dem Deckblatt eines Eingewöhnungskonzepts sowie die abschließende komparative Analyse wird die spezifische Art der Herstellung und Kontextualisierung der Bilder herausgearbeitet. Deutlich werden die Wissensbestände, die in den Konzepten eine bedeutsame Rolle spielen und wie sie bei der Thematisierung der Eingewöhnung – z. B. gegenüber den Eltern – zur Geltung gebracht werden. DOI 10.18442/09

Immunostimulation by OX40 Ligand Transgenic Ewing Sarcoma Cells

Interleukin-2 (IL-2) transgenic Ewing sarcoma cells can induce tumor specific T and NK cell responses and reduce tumor growth in vivo and in vitro. Nevertheless, the efficiency of this stimulation is not high enough to inhibit tumor growth completely. In addition to recognition of the cognate antigen, optimal T cell stimulation requires signals from so-called co-stimulatory molecules. Several members of the tumor necrosis factor superfamily (TNFSF) have been identified as co-stimulatory molecules that can augment anti-tumor immune responses. OX40 (CD134) and OX40 ligand (OX40L = CD252; also known as tumor necrosis factor ligand family member 4) is one example for such receptor/ligand pair with co-stimulatory function. In the present investigation we generated OX40L transgenic Ewing sarcoma cells and tested their immuno-stimulatory activity in vitro. OX40L transgenic Ewing sarcoma cells showed preserved expression of Ewing sarcoma associated (anti)gens including lipase member I (LIPI), cyclin D1 (CCND1), cytochrome P450 family member 26B1 (CYP26B1) and the Ewing sarcoma breakpoint region 1-friend leukemia virus integration 1 (EWSR1-FLI1) oncogene. OX40L expressing tumor cells showed a trend for enhanced immune stimulation against Ewing sarcoma cells in combination with IL-2 and stimulation of CD137. Our data suggest that inclusion of the OX40/OX40L pathway of co-stimulation might improve immunotherapy strategies for treatment of Ewing sarcoma

Immunogenicity of the Envelope Surface Unit of Human Endogenous Retrovirus K18 in Mice

The triggers for the development of multiple sclerosis (MS) have not been fully understood to date. One hypothesis proposes a viral etiology. Interestingly, viral proteins from human endogenous retroviruses (HERVs) may play a role in the pathogenesis of MS. Allelic variants of the HERV-K18 env gene represent a genetic risk factor for MS, and the envelope protein is considered to be an Epstein–Barr virus-trans-activated superantigen. To further specify a possible role for HERV-K18 in MS, the present study examined the immunogenicity of the purified surface unit (SU). HERV-K18(SU) induced envelope-specific plasma IgG in immunized mice and triggered proliferation of T cells isolated from these mice. It did not trigger phenotypic changes in a mouse model of experimental autoimmune encephalomyelitis. Further studies are needed to investigate the underlying mechanisms of HERV-K18 interaction with immune system regulators in more detail

Oxidative damage control in a human (mini-) organ: Nrf2 activation protects against oxidative stress-induced hair growth inhibition

The in situ control of redox insult in human organs is of major clinical relevance, yet remains incompletely understood. Activation of Nrf2, the “master regulator” of genes controlling cellular redox homeostasis, is advocated as a therapeutic strategy for diseases with severely impaired redox balance. It remains to be shown whether this strategy is effective in human organs, rather than isolated human cell types. We have therefore explored the role of Nrf2 in a uniquely accessible human (mini-) organ, human scalp hair follicles (HFs). Microarray and qPCR analysis of human HFs following Nrf2 activation using sulforaphane identified the modulation of phase II metabolism, ROS clearance, the pentose phosphate pathway and glutathione homeostasis. Nrf2 knockdown (siRNA) in cultured human HFs confirmed the regulation of key Nrf2 target genes (i.e. HO-1, NQO1, GSR, GCLC, ABCC1, PRDX1). Importantly, Nrf2 activation significantly reduced ROS levels and associated lipid peroxidation. Nrf2 pre-activation reduced oxidative stress-stimulated (H2O2 or menadione) premature catagen and hair growth inhibition, significantly ameliorated the H2O2-dependent increase in matrix keratinocyte apoptosis and reversed the ROS-induced reduction in proliferation. This study thus provides direct evidence for the crucial role of Nrf2 in protecting human organ function (i.e. scalp HFs) against redox insult

Knocking out C9ORF72 Exacerbates Axonal Trafficking Defects Associated with Hexanucleotide Repeat Expansion and Reduces Levels of Heat Shock Proteins

In amyotrophic lateral sclerosis (ALS) motor neurons (MNs) undergo dying-back, where the distal axon degenerates before the soma. The hexanudeotide repeat expansion (HRE) in C9ORF72 is the most common genetic cause of ALS, but the mechanism of pathogenesis is largely unknown with both gain- and loss-of-function mechanisms being proposed. To better understand C9ORF72-ALS pathogenesis, we generated isogenic induced pluripotent stem cells. MNs with HRE in C9ORF72 showed decreased axonal trafficking compared with gene corrected MNs. However, knocking out C9ORF72 did not recapitulate these changes in MNs from healthy controls, suggesting a gain-of-function mechanism. In contrast, knocking out C9ORF72 in MNs with HRE exacerbated axonal trafficking defects and increased apoptosis as well as decreased levels of HSP70 and HSP40, and inhibition of HSPs exacerbated ALS phenotypes in MNs with HRE. Therefore, we propose that the HRE in C9ORF72 induces ALS pathogenesis via a combination of gain- and loss-of-function mechanisms

Generation of four iPSC lines from four patients with Leigh syndrome carrying homoplasmic mutations m.8993T > G or m.8993T > C in the mitochondrial gene MT-ATP6.

We report the generation of four human iPSC lines (8993-A12, 8993-B12, 8993-C11, and 8993-D7) from fibroblasts of four patients affected by maternally inherited Leigh syndrome (MILS) carrying homoplasmic mutations m.8993T > G or m.8993T > C in the mitochondrial gene MT-ATP6. We used Sendai viruses to deliver reprogramming factors OCT4, SOX2, KLF4, and c-MYC. The established iPSC lines expressed pluripotency markers, exhibited a normal karyotype, were capable to form cells of the three germ layers in vitro, and retained the MT-ATP6 mutations at the same homoplasmic level of the parental fibroblasts

WebHERV: A Web Server for the Computational Investigation of Gene Expression Associated With Endogenous Retrovirus-Like Sequences

More than eight percent of the human genome consists of human endogenous retroviruses (HERVs). Typically, the expression of HERVs is repressed, but varying activities of HERVs have been observed in diseases ranging from cancer to neuro-degeneration. Such activities can include the transcription of HERV-derived open reading frames, which can be translated into proteins. However, as a consequence of mutations that disrupt open reading frames, most HERV-like sequences have lost their protein-coding capacity. Nevertheless, these loci can still influence the expression of adjacent genes and, hence, mediate biological effects. Here, we present WebHERV (http://calypso.informatik.uni-halle.de/WebHERV/), a web server that enables the computational prediction of active HERV-like sequences in the human genome based on a comparison of genome coordinates of expressed sequences uploaded by the user and genome coordinates of HERV-like sequences stored in the specialized key-value store DRUMS. Using WebHERV, we predicted putative candidates of active HERV-like sequences in Hodgkin lymphoma (HL) cell lines, validated one of them by a modified SMART (switching mechanism at 5′ end of RNA template) technique, and identified a new alternative transcription start site for cytochrome P450, family 4, subfamily Z, polypeptide 1 (CYP4Z1)

The biogenesis and characterization of mammalian microRNAs of mirtron origin

Mirtrons, short hairpin pre-microRNA (miRNA) mimics directly produced by intronic splicing, have recently been identified and experimentally confirmed in invertebrates. While there is evidence to suggest several mammalian miRNAs have mirtron origins, this has yet to be experimentally demonstrated. Here, we characterize the biogenesis of mammalian mirtrons by ectopic expression of splicing-dependent mirtron precursors. The putative mirtrons hsa-miR-877, hsa-miR-1226 and mmu-miR-1224 were designed as introns within eGFP. Correct splicing and function of these sequences as introns was shown through eGFP fluorescence and RT–PCR, while all mirtrons suppressed perfectly complementary luciferase reporter targets to levels similar to that of corresponding independently expressed pre-miRNA controls. Splicing-deficient mutants and disruption of key steps in miRNA biogenesis demonstrated that mirtron-mediated gene knockdown was splicing-dependent, Drosha-independent and had variable dependence on RNAi pathway elements following pre-miRNA formation. The silencing effect of hsa-miR-877 was further demonstrated to be mediated by the generation of short anti-sense RNA species expressed with low abundance. Finally, the mammalian mirtron hsa-miR-877 was shown to reduce mRNA levels of an endogenous transcript containing hsa-miR-877 target sites in neuronal SH-SY5Y cells. This work confirms the mirtron origins of three mammalian miRNAs and suggests that they are a functional class of splicing-dependent miRNAs which are physiologically active