Quantification of global DNA methylation with infrared fluorescence in liver and muscle tissues of differentially fed boars

Methylation of cytosine residues at CpG sites is involved in various biological processes to control gene regulation and gene expression. Global DNA methylation is changed in different tumors and in cloned animals. Global DNA methylation can be accurately quantified by dot blot analysis with infrared (IR) fluorophores. Methylated lambda DNA was used as model DNA to develop and validate an immunochemical assay with IR fluorescence detection. Two different IR fluorophores were used, one to detect 5-methylcytosine and another to account for DNA loading. A sensitive infrared detection method was established which is suitable for accurate and reproducible quantification of global DNA methylation across a wide dynamic range. This method was subsequently employed to quantify global DNA methylation in liver and in muscle tissues of boars which have received either a control diet or a methyl supplemented diet in an ongoing study. A significant difference in global DNA methylation is indicated in muscle but not in liver tissue between the two groups of boars.

The bovine dilated cardiomyopathy locus maps to a 1.0-Mb interval on chromosome 18

Cardiomyopathies are myocardial diseases that lead to cardiac dysfunction, heart failure, arrhythmia, and sudden death. In human medicine, cardiomyopathies frequently warrant heart transplantation in children and adults. Bovine dilated cardiomyopathy (BDCMP) is a heart muscle disorder that has been observed during the last 30 years in cattle of Holstein-Friesian origin. In Switzerland BDCMP affects Swiss Fleckvieh and Red Holstein breeds. BDCMP is characterized by a cardiac enlargement with ventricular remodeling and chamber dilatation. The common symptoms in affected animals are subacute subcutaneous edema, congestion of the jugular veins, and tachycardia with gallop rhythm. A cardiomegaly with dilatation and hypertrophy of all heart chambers, myocardial degeneration, and fibrosis are typical postmortem findings. It was shown that all BDCMP cases reported worldwide traced back to a red factor-carrying Holstein-Friesian bull, ABC Reflection Sovereign. An autosomal recessive mode of inheritance was proposed for BDCMP. Recently, the disease locus was mapped to a 6.7-Mb interval MSBDCMP06-BMS2785 on bovine Chr 18 (BTA18). In the present study the BDCMP locus was fine mapped by using a combined strategy of homozygosity mapping and association study. A BAC contig of 2.9 Mb encompassing the crucial interval was constructed to establish the correct marker order on BTA18. We show that the disease locus is located in a gene-rich interval of 1.0 Mb and is flanked by the microsatellite markers DIK3006 and MSBDCMP51.

DNA methylation patterns at the IGF2-H19 locus in sperm of Swiss Landrace and Swiss Large White boars

DNA methylation patterns at the IGF2-H19 locus were investigated in sperm DNA from Swiss Landrace (SL) and Swiss Large White (LW) boars. The putative IGF2 differentially methylated regions (DMR) 0, 1 and 2, a quantitative trait nucleotide (QTN) region in the intron 3 and a CpG island in the intron 4 of the IGF2 gene as well as three regions around porcine CTCF binding sites within the H19 differentially methylated domain (DMD) were selected for the DNA methylation analysis. In both breeds putative IGF2 DMR0, 1, 2 and H19 DMD were hypermethylated. Significant differences in DNA methylation content were found between the two breeds in the two DMD regions proximal to the H19 gene. The IGF2 QTN region and the CpG island in the IGF2 intron 4 were hypomethylated in sperm DNA of both breeds. The methylation analysis revealed significantly more methylated CpG sites in the intron 4 of sperm from the LW breed than in that from SL. No difference was found in global DNA methylation between the two breeds. These results indicate differences in DNA methylation patterns between breeds and it remains to be established whether variation in DNA methylation patterns impacts on phenotypic traits.

Changes in German sport participation

Sport has become a highly differentiated social phenomenon in recent years. Changes in society, such as individualization, the growing significance of the health and body culture, and changing values, are considered to be generative mechanisms for increasing social importance and the differentiation of modern sport. Although discussions in sport sociology attribute the changes observed in recent decades of sport participation to a socially determined differentiation of sport, this premise has hardly ever been empirically tested. The present study examines to what extent the postulated developments in sport can be observed on the micro level of those engaging in sport, by examining sport behaviour from a contemporary historical perspective. Based on a life-course approach to research, a total of 1739 over 50-year-olds in Germany were asked about their sport participation as part of a retrospective longitudinal study. Results show that the increasing differentiation of sport can be documented by more diversified forms of individual sport careers. During a 30-year observation period the popularity of competitive sport decreased and the variety of ways in which sport was organized increased. A differentiated analysis based on examining three birth cohorts showed that the reported change in sport participation can be attributed to age, cohort and period effects. In addition, the present study examines how specific events in contemporary history are reflected in individual sporting careers. Sport careers in Chemnitz (Eastern Germany) and Braunschweig (Western Germany) differed before German reunification, but these differences have evened out after the political changes and the process of transformation.

In search of epigenetic marks in testes and sperm cells of differentially fed boars

In search of transmittable epigenetic marks we investigated gene expression in testes and sperm cells of differentially fed F0 boars from a three generation pig feeding experiment that showed phenotypic differences in the F2 generation. RNA samples from 8 testes of boars that received either a diet enriched in methylating micronutrients or a control diet were analyzed by microarray analysis. We found moderate differential expression between testes of differentially fed boars with a high FDR of 0.82 indicating that most of the differentially expressed genes were false positives. Nevertheless, we performed a pathway analysis and found disparate pathway maps of development_A2B receptor: action via G-protein alpha s, cell adhesion_Tight junctions and cell adhesion_Endothelial cell contacts by junctional mechanisms which show inconclusive relation to epigenetic inheritance. Four RNA samples from sperm cells of these differentially fed boars were analyzed by RNA-Seq methodology. We found no differential gene expression in sperm cells of the two groups (adjusted P-value>0.05). Nevertheless, we also explored gene expression in sperm by a pathway analysis showing that genes were enriched for the pathway maps of bacterial infections in cystic fibrosis (CF) airways, glycolysis and gluconeogenesis p.3 and cell cycle_Initiation of mitosis. Again, these pathway maps are miscellaneous without an obvious relationship to epigenetic inheritance. It is concluded that the methylating micronutrients moderately if at all affects RNA expression in testes of differentially fed boars. Furthermore, gene expression in sperm cells is not significantly affected by extensive supplementation of methylating micronutrients and thus RNA molecules could not be established as the epigenetic mark in this feeding experiment.

Aktive Online-Nachsorge für Patienten mit Schizophrenie-Diagnose? Bedarf und Risiken aus Sicht der Behandler

Kognitive Interventionen haben sich bei der Behandlung von Schizophrenie als wirksam erwiesen und werden mittlerweile in nationalen Richtlinien empfohlen. Viele Patienten mit Schizophrenie-Diagnose haben jedoch keinen Zugang zu post-stationären, ambulanten Psychotherapie-Plätzen nach stationären Klinikaufenthalten. Eine Möglichkeit, Patienten den Zugang zu kognitiven Interventionen zu ermöglichen, sind Online-Nachsorge-Programme. Die vorliegende Befragung hatte zum Ziel, den Bedarf sowie mögliche Risiken und Nachteile von Online-Nachsorge für Schizophrenie aus Sicht der Behandler zu erfassen. Dreißig psychologische und ärztliche BehandlerInnen von Patienten mit Schizophrenie-Diagnose nahmen an einer Online-Befragung teil (Alter: M=34.5 Jahre, SD=9.8; 24 bis 60 Jahre; 63% weiblich; 70% PsychologInnen; 73% in Weiterbildung Psychotherapie, 20% approbiert). Die Stichprobe hatte im Mittel M=6.2 Jahre Berufserfahrung (SD=7.5) und war zum Großteil bei 26 bis 100 Behandlungen von Patienten mit Schizophrenie-Diagnose involviert. Im Mittel erhält die Hälfte der Patienten nach Einschätzung der Behandler keine post-stationäre psychotherapeutische Nachsorge mit nachgewiesen wirksamen und hilfreichen psychotherapeutischen Verfahren (M=50.2%, SD=38.2%). Die Behandler befürworteten die Online-Nachsorge bei Patienten, die keinen ambulanten Therapieplatz zur Verfügung haben, ohne weiteres (53.3%) oder zumindest nach wissenschaftlicher Überprüfung auf Unbedenklichkeit und Wirksamkeit (46.7%). Die nicht-repräsentative Online-Erhebung mit vor allem jungen, in Ausbildung befindlichen BehandlerInnen legt den Schluss nahe, dass Online-Nachsorge mit aktiven Interventionen für Patienten mit Schizophrenie-Diagnose aus Sicht der Behandler prinzipiell vertretbar und erwünscht ist. Mögliche Voraussetzungen (wie z.B. Medikation) für die Teilnahme an Online-Nachsorge-Programmen sowie mögliche Risiken aus Sicht von BehandlerInnen werden diskutiert.

Reevaluation of the MASTER-2009 MLI and H-10 Debris Modeling

The current paper is an excerpt from the doctoral thesis ”Multi-Layer Insulation as Contribution to Orbital Debris”written at the Institute of Aerospace Systems of the Technische Universit ̈at of Braunschweig. The Multi-Layer In-sulation (MLI) population included in ESA’s MASTER-2009 (M eteoroid and Space-Debris Terrestrial Environment Reference) software is based on models for two mechanisms: One model simulates the release of MLI debris during fragmentation events while another estimates the continuo us release of larger MLI pieces due to aging related deterioration of the material. The aim of the thesis was to revise the MLI models from the base up followed by a re-validation of the simulated MLI debris population. The validation is based on comparison to measurement data of the GEO and GTO debris environment obtained by the Astronomical Institute of the University of Bern (AIUB) using ESA’s Space Debris Telescope (ESASDT), the 1-m Zeiss telescope located at the Optical Ground Station (OGS) at the Teide Observatory at Tenerife, Spain. The re-validation led to the conclusion that MLI may cover a much smaller portion of the observed objects than previously published. Further investigation of the resulting discrepancy revealed that the contribution of altogether nine known Ariane H-10 upper stage explosion events which occurred between 1984 and 2002 has very likely been underestimated in past simulations.

Sensibilisierung und Motivierung von Lehrenden für eine Kompetenzorientierung in der Lehre

Handreichung Lernergebnisse Teil 2: Anwendungsbeispiele einer outcomeorientierten Programmentwicklung. Eine Publikation der wissenschaftlichen Begleitung des Bund-Länder-Wettbewerbs "Aufstieg durch Bildung: offene Hochschulen". Berlin.

Exact Unification and Admissibility

A new hierarchy of "exact" unification types is introduced, motivated by the study of admissible rules for equational classes and non-classical logics. In this setting, unifiers of identities in an equational class are preordered, not by instantiation, but rather by inclusion over the corresponding sets of unified identities. Minimal complete sets of unifiers under this new preordering always have a smaller or equal cardinality than those provided by the standard instantiation preordering, and in significant cases a dramatic reduction may be observed. In particular, the classes of distributive lattices, idempotent semigroups, and MV-algebras, which all have nullary unification type, have unitary or finitary exact type. These results are obtained via an algebraic interpretation of exact unification, inspired by Ghilardi's algebraic approach to equational unification.

Evidence for two protein coding transcripts at the Igf2as locus

The insulin-like growth factor 2 antisense (Igf2as) gene is part of the Ins-Igf2-H19 imprinted gene cluster. The function of the paternally expressed Igf2as is still elusive. In our previous work, we showed that Igf2as transcripts were located in the cytoplasm of C2C12 mouse myoblast cells, associated with polysomes and polyadenylated suggesting that Igf2as is protein coding. In the present work, the protein coding capacity of Igf2as was investigated. We demonstrate for the first time the existence of a polypeptide translated from an Igf2as construct. Furthermore, an RNA-Seq analysis was performed using RNA prepared from skeletal muscles of newborn wild-type and ∆ DMR1-U2 mice to further elucidate the function of Igf2as transcripts. We found no evidence for a regulatory role of Igf2as in the imprinted gene cluster. Interestingly, the RNA-Seq analysis indicated that Igf2as plays a role in the energy metabolism, the cell cycle, histone acetylation and muscle contraction pathways. Our Igf2as investigations further elucidated that there are two distinct Igf2as transcripts corresponding to two putative ORFs.