@article{SbirkovKwokBhamraetal.2017,
  author    = {Sbirkov, Yordan and Kwok, Colin and Bhamra, Amandeep and Thompson, Andrew J. and Gil, Veronica and Zelent, Arthur and Petrie, Kevin},
  title     = {Semi-quantitative mass spectrometry in AML cells identifies new non-genomic targets of the EZH2 methyltransferase},
  series = {International Journal of Molecular Sciences},
  volume    = {18},
  journal   = {International Journal of Molecular Sciences},
  number    = {7},
  issn      = {1422-0067},
  doi       = {10.3390/ijms18071440},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-285541},
  year      = {2017},
  abstract  = {Alterations to the gene encoding the EZH2 (KMT6A) methyltransferase, including both gain-of-function and loss-of-function, have been linked to a variety of haematological malignancies and solid tumours, suggesting a complex, context-dependent role of this methyltransferase. The successful implementation of molecularly targeted therapies against EZH2 requires a greater understanding of the potential mechanisms by which EZH2 contributes to cancer. One aspect of this effort is the mapping of EZH2 partner proteins and cellular targets. To this end we performed affinity-purification mass spectrometry in the FAB-M2 HL-60 acute myeloid leukaemia (AML) cell line before and after all-trans retinoic acid-induced differentiation. These studies identified new EZH2 interaction partners and potential non-histone substrates for EZH2-mediated methylation. Our results suggest that EZH2 is involved in the regulation of translation through interactions with a number of RNA binding proteins and by methylating key components of protein synthesis such as eEF1A1. Given that deregulated mRNA translation is a frequent feature of cancer and that eEF1A1 is highly expressed in many human tumours, these findings present new possibilities for the therapeutic targeting of EZH2 in AML.},
  language  = {en}
}
@phdthesis{Reichenbach2020,
  author    = {Reichenbach, Juliane Renate},
  title     = {Paternal age effects on sperm DNA methylation and its impact on the next generation},
  doi       = {10.25972/OPUS-19980},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-199805},
  school      = {Universit{\"a}t W{\"u}rzburg},
  year      = {2020},
  abstract  = {The effect of late parenthood on the offspring´s physical and mental health status has recently become an increasingly important topic of discussion. Studies on neurodevelopmental disorders in children of older parents (Naserbakht et al., 2011) outline the negative consequences of aging fathers as unpredictable compared to the better-understood unfavorable maternal influences (Cedars et al. 2015). This may be due to the fact that lifelong production of male gametes becomes more susceptible to error, not only for somatic mutations. Non-genomic mechanisms such as epigenetic methylation also alter DNA dynamically throughout life (Jones et al., 2015) and influence the aging human sperm DNA (Jenkins et al., 2014). These methylation changes may be transmitted to the next generation via epigenetic inheritance mechanisms (Milekic et al., 2015), which may negatively impact the sensitive epigenetic regulation of cell differentiation in the embryonic period (Curley et al., 2011; Spiers et al., 2015). Accordingly, Nardone et al. (2014) reported several hypomethylated regions in autistic patients, illustrating potential epigenetic influences on the multifactorial pathogenesis of neuropsychiatric disorders. In the present study, the methylation status of five gene regions in the sperm DNA of males of different ages was analyzed by two techniques - pyrosequencing and deep bisulfite sequencing. Two gene regions, FOXK1 and DMPK, showed a highly significant age-related methylation loss and FOXK1 a reduced methylation variation at the level of single alleles. In addition, the examined gene region of FOXK1 showed significant methylation changes in the fetal cord blood DNA of the respective offspring of the sperm donor. This fact suggests a transfer of age-related methylation loss to the next generation. Interestingly, a methylation analysis at the level of single alleles showed that the methylation loss was inherited exclusively by the father. FOXK1 is a transcription factor that plays an important role in the epigenetic regulation of the cell cycle during embryonic neuronal development (Huang et al., 2004; Wijchers et al., 2006). For this reason, the methylation status of FOXK1 in the blood of autistic patients and an age- and sex-matched control group was investigated. While both groups showed age-associated FOXK1 methylation loss, a faster dynamics of methylation change was observed in the autistic group. Although further studies are needed to uncover inheritance mechanisms of epigenetic information, the present results show an evident influence of age-related methylation changes on offspring. When advising future fathers, it is important to consider how the paternal epigenome is altered by aging and can have a negative impact on the developing embryo.},
  subject      = {Epigenetik},
  language  = {en}
}
@article{HurdGruebelWojciechowskietal.2021,
  author    = {Hurd, Paul J. and Gr{\"u}bel, Kornelia and Wojciechowski, Marek and Maleszka, Ryszard and R{\"o}ssler, Wolfgang},
  title     = {Novel structure in the nuclei of honey bee brain neurons revealed by immunostaining},
  series = {Scientific Reports},
  volume    = {11},
  journal   = {Scientific Reports},
  doi       = {10.1038/s41598-021-86078-5},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-260059},
  pages     = {6852},
  year      = {2021},
  abstract  = {In the course of a screen designed to produce antibodies (ABs) with affinity to proteins in the honey bee brain we found an interesting AB that detects a highly specific epitope predominantly in the nuclei of Kenyon cells (KCs). The observed staining pattern is unique, and its unfamiliarity indicates a novel previously unseen nuclear structure that does not colocalize with the cytoskeletal protein f-actin. A single rod-like assembly, 3.7-4.1 mu m long, is present in each nucleus of KCs in adult brains of worker bees and drones with the strongest immuno-labelling found in foraging bees. In brains of young queens, the labelling is more sporadic, and the rod-like structure appears to be shorter (similar to 2.1 mu m). No immunostaining is detectable in worker larvae. In pupal stage 5 during a peak of brain development only some occasional staining was identified. Although the cellular function of this unexpected structure has not been determined, the unusual distinctiveness of the revealed pattern suggests an unknown and potentially important protein assembly. One possibility is that this nuclear assembly is part of the KCs plasticity underlying the brain maturation in adult honey bees. Because no labelling with this AB is detectable in brains of the fly Drosophila melanogaster and the ant Camponotus floridanus, we tentatively named this antibody AmBNSab (Apis mellifera Brain Neurons Specific antibody). Here we report our results to make them accessible to a broader community and invite further research to unravel the biological role of this curious nuclear structure in the honey bee central brain.},
  language  = {en}
}
@article{AhmadWolberEckardtetal.2012,
  author    = {Ahmad, Ruhel and Wolber, Wanja and Eckardt, Sigrid and Koch, Philipp and Schmitt, Jessica and Semechkin, Ruslan and Geis, Christian and Heckmann, Manfred and Br{\"u}stle, Oliver and McLaughlin, John K. and Sir{\´e}n, Anna-Leena and M{\"u}ller, Albrecht M.},
  title     = {Functional Neuronal Cells Generated by Human Parthenogenetic Stem Cells},
  series = {PLoS One},
  volume    = {7},
  journal   = {PLoS One},
  number    = {8},
  doi       = {10.1371/journal.pone.0042800},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-130268},
  pages     = {e42800},
  year      = {2012},
  abstract  = {Parent of origin imprints on the genome have been implicated in the regulation of neural cell type differentiation. The ability of human parthenogenetic (PG) embryonic stem cells (hpESCs) to undergo neural lineage and cell type-specific differentiation is undefined. We determined the potential of hpESCs to differentiate into various neural subtypes. Concurrently, we examined DNA methylation and expression status of imprinted genes. Under culture conditions promoting neural differentiation, hpESC-derived neural stem cells (hpNSCs) gave rise to glia and neuron-like cells that expressed subtype-specific markers and generated action potentials. Analysis of imprinting in hpESCs and in hpNSCs revealed that maternal-specific gene expression patterns and imprinting marks were generally maintained in PG cells upon differentiation. Our results demonstrate that despite the lack of a paternal genome, hpESCs generate proliferating NSCs that are capable of differentiation into physiologically functional neuron-like cells and maintain allele-specific expression of imprinted genes. Thus, hpESCs can serve as a model to study the role of maternal and paternal genomes in neural development and to better understand imprinting-associated brain diseases.},
  language  = {en}
}
@article{AlmanzarKleinSchmalzingetal.2016,
  author    = {Almanzar, Giovanni and Klein, Matthias and Schmalzing, Marc and Hilligardt, Deborah and El Hajj, Nady and Kneitz, Hermann and Wild, Vanessa and Rosenwald, Andreas and Benoit, Sandrine and Hamm, Henning and Tony, Hans-Peter and Haaf, Thomas and Goebeler, Matthias and Prelog, Martina},
  title     = {Disease Manifestation and Inflammatory Activity as Modulators of Th17/Treg Balance and RORC/FoxP3 Methylation in Systemic Sclerosis},
  series = {International Archives of Allergy and Immunology},
  volume    = {171},
  journal   = {International Archives of Allergy and Immunology},
  number    = {2},
  issn      = {1018-2438},
  doi       = {10.1159/000450949},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-196577},
  pages     = {141-154},
  year      = {2016},
  abstract  = {Background: There is much evidence that T cells are strongly involved in the pathogenesis of localized and systemic forms of scleroderma (SSc). A dysbalance between FoxP3+ regulatory CD4+ T cells (Tregs) and inflammatory T-helper (Th) 17 cells has been suggested. Methods: The study aimed (1) to investigate the phenotypical and functional characteristics of Th17 and Tregs in SSc patients depending on disease manifestation (limited vs. diffuse cutaneous SSc, dcSSc) and activity, and (2) the transcriptional level and methylation status of Th17- and Treg-specific transcription factors. Results: There was a concurrent accumulation of circulating peripheral IL-17-producing CCR6+ Th cells and FoxP3+ Tregs in patients with dcSSc. At the transcriptional level, Th17- and Treg-associated transcription factors were elevated in SSc. A strong association with high circulating Th17 and Tregs was seen with early, active, and severe disease presentation. However, a diminished suppressive function on autologous lymphocytes was found in SSc-derived Tregs. Significant relative hypermethylation was seen at the gene level for RORC1 and RORC2 in SSc, particularly in patients with high inflammatory activity. Conclusions: Besides the high transcriptional activity of T cells, attributed to Treg or Th17 phenotype, in active SSc disease, Tregs may be insufficient to produce high amounts of IL-10 or to control proliferative activity of effector T cells in SSc. Our results suggest a high plasticity of Tregs strongly associated with the Th17 phenotype. Future directions may focus on enhancing Treg functions and stabilization of the Treg phenotype.},
  language  = {en}
}
@phdthesis{Berking2021,
  author    = {Berking, Ann-Cathrine},
  title     = {Assoziationsuntersuchung von ausgew{\"a}hlten Polymorphismen der Gene DNMT3A und DNMT3B mit der Panikst{\"o}rung},
  doi       = {10.25972/OPUS-23468},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-234687},
  school      = {Universit{\"a}t W{\"u}rzburg},
  year      = {2021},
  abstract  = {Currently, the vulnerability-stress model, in the sense of a multifactorial explanatory model, is considered to be the most appropriate to represent the etiopathogenesis of anxiety disorders. Epigenetic mechanisms are understood as a bridge between genetic factors and environmental factors. This includes the methylation of specific DNA regions, which is mediated by DNA methyltransferases. These enzymes have rarely been the focus of psychiatric research in relation to anxiety disorders. Therefore, this work deals with selected single nucleotide polymorphisms of the DNMT3A and DNMT3B gene and investigates whether these SNPs and/or their haplotypes are associated panic disorder and/or with dimensional psychological characteristics, such as anxiety-related cognition or anxiety sensitivity. In summary, a significant or nominally significant association of two SNPs with anxiety-related characteristics such was shown. To better assess these associations, replications with sufficient test strength are required . Given the demonstrated association with PSWQ, investigation of another anxiety phenotype, Generalized Anxiety Disorder, is also sensible. As a further step, the functionality of the significantly associated SNPs should be performed. In addition, another DNMT, Dnmt1, is associated with fear conditioning, and the methylation patterns of the DNMTs themselves also appear to have an impact on the development of anxiety disorders. Therefore, an investigation of the DNMT1 gene and the methylation patterns of the DNMT genes are further reasonable steps to better understand a possible influence of DNMTs on the development of anxiety disorders and on anxiety-related psychological characteristics.},
  language  = {de}
}
@phdthesis{Jonas2008,
  author    = {Jonas, Ren{\´e}},
  title     = {Arsen-induzierte Zyto- und Gentoxizit{\"a}t sowie deren Modulation},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-28772},
  school      = {Universit{\"a}t W{\"u}rzburg},
  year      = {2008},
  abstract  = {Arsen ist daf{\"u}r bekannt, dass es mutagen und kanzerogen wirkt und ein gentoxisches Potential besitzt. Die Mechanismen, durch die diese Effekte ausge{\"u}bt werden, sind noch nicht vollst{\"a}ndig aufgekl{\"a}rt. Es konnte jedoch gezeigt werden, dass Parameter, die mit der Freisetzung reaktiver Sauerstoffspezies (ROS), z.B. Superoxiddismutaseaktivit{\"a}t und H{\"a}moxygenase-Genexpression, und Ver{\"a}nderungen des epigenetischen Musters der DNA, z.B. Depletion von S-Adenosylmethionin, in Zusammenhang stehen, durch Arsen beeinflusst werden. In dieser Studie wurde versucht, das gentoxische Potential von Arsen mit Hilfe des Comet Assay, eines Standard-Gentoxizit{\"a}tstests, zu charakterisieren sowie zu pr{\"u}fen, ob dieser Test eine geeignete Messmethode f{\"u}r die gentoxische Wirkung von Arsen darstellt. Dies wurde unter Heranziehung verschiedener additiver Messgr{\"o}ßen wie der Vitalit{\"a}t und der Proliferation sowie der parallelen Quantifizierung der Mitose-, C-Mitose-, Mikrokern- und Apoptosefrequenzen der verwendeten murinen L5178Y-Zellen durchgef{\"u}hrt. Des Weiteren wurde der den Arsen-bedingten DNA-Sch{\"a}den zugrundeliegende Mechanismus genauer beleuchtet. Unter Zuhilfenahme verschiedener Modulatoren wurden durch Arsen induzierter oxidativer Stress und durch Arsen induzierte Ver{\"a}nderung der epigenetischen DNA-Struktur untersucht. Ferner wurde gepr{\"u}ft, inwieweit die Inhibition von oxidativem Stress und Hypomethylierung der DNA zur Verringerung von potenziellen Folgen wie der Entstehung unnat{\"u}rlicher Mitosemorphologien und chromosomaler Aberrationen beitragen k{\"o}nnen, die wiederum eventuell in der Entstehung von Karzinomen resultieren k{\"o}nnen. F{\"u}r die Modulation der Freisetzung von ROS wurden als prooxidative Substanz 4-Nitrochinolin-1-Oxid und als Antioxidantien Benfotiamin (Vitamin-B1-Prodrug), N-Acetylcystein (NAC) und \&\#945;-Tocopherol (Vitamin E) ausgew{\"a}hlt. Das Methylierungs¬muster der DNA sollte durch das hypomethylierende Agens 5-Azacytidin und durch die potenziell hypermethylierenden Verbindungen S-Adenosylmethionin (SAM) und Folat beeinflusst werden. Die Untersuchungen bez{\"u}glich des gentoxischen Potentials von Arsen und die Eignung des Comet Assay f{\"u}r dessen Quantifizierung ergaben, dass unter Miteinbeziehung der erw{\"a}hnten additiven Parameter und der Quantifizierung nach Behandlung mit unterschiedlichen Arsen-Konzentrationen nach unterschiedlich langen Behandlungszeiten die im Comet Assay erzielten Werte als korrekt und zuverl{\"a}ssig angesehen werden k{\"o}nnen. Des Weiteren zeigten die Untersuchungen der Freisetzung von ROS und der Ver{\"a}nderung des DNA-Methylierungsmusters mit Hilfe von Modulatoren, dass beide Mechanismen an den Arsen-induzierten Effekten beteiligt sind. Nicht nur konnte mit Hilfe der Modulatoren jeweils die Inhibition der Freisetzung von ROS und der DNA-Hypomethylierung erreicht werden, es konnte zudem gezeigt werden, dass die Substanzen auch die Reduktion der erh{\"o}hten Anzahl unnat{\"u}rlicher Mitosemorphologien und chromosomaler Aberrationen bewirkten. Dieser Zusammenhang konnte in dieser Studie zum ersten Mal aufgezeigt werden und k{\"o}nnte im Hinblick auf die potenzielle Erniedrigung der Krebsinzidenzen durch Supplementierung der Bev{\"o}lkerung in Gebieten mit Arsen-belastetem Trinkwasser mit den genannten Modulatoren von Bedeutung sein.},
  subject      = {Oxidativer Stress},
  language  = {de}
}
@article{MaierhoferFlunkertDittrichetal.2017,
  author    = {Maierhofer, Anna and Flunkert, Julia and Dittrich, Marcus and M{\"u}ller, Tobias and Schindler, Detlev and Nanda, Indrajit and Haaf, Thomas},
  title     = {Analysis of global DNA methylation changes in primary human fibroblasts in the early phase following X-ray irradiation},
  series = {PLoS ONE},
  volume    = {12},
  journal   = {PLoS ONE},
  number    = {5},
  doi       = {10.1371/journal.pone.0177442},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-170895},
  pages     = {e0177442},
  year      = {2017},
  abstract  = {Epigenetic alterations may contribute to the generation of cancer cells in a multi-step process of tumorigenesis following irradiation of normal body cells. Primary human fibroblasts with intact cell cycle checkpoints were used as a model to test whether X-ray irradiation with 2 and 4 Gray induces direct epigenetic effects (within the first cell cycle) in the exposed cells. ELISA-based fluorometric assays were consistent with slightly reduced global DNA methylation and hydroxymethylation, however the observed between-group differences were usually not significant. Similarly, bisulfite pyrosequencing of interspersed LINE-1 repeats and centromeric α-satellite DNA did not detect significant methylation differences between irradiated and non-irradiated cultures. Methylation of interspersed ALU repeats appeared to be slightly increased (one percentage point; p = 0.01) at 6 h after irradiation with 4 Gy. Single-cell analysis showed comparable variations in repeat methylation among individual cells in both irradiated and control cultures. Radiation-induced changes in global repeat methylation, if any, were much smaller than methylation variation between different fibroblast strains. Interestingly, α-satellite DNA methylation positively correlated with gestational age. Finally, 450K methylation arrays mainly targeting genes and CpG islands were used for global DNA methylation analysis. There were no detectable methylation differences in genic (promoter, 5' UTR, first exon, gene body, 3' UTR) and intergenic regions between irradiated and control fibroblast cultures. Although we cannot exclude minor effects, i.e. on individual CpG sites, collectively our data suggest that global DNA methylation remains rather stable in irradiated normal body cells in the early phase of DNA damage response.},
  language  = {en}
}
@article{WenzArndtSamnick2022,
  author    = {Wenz, Jan and Arndt, Felix and Samnick, Samuel},
  title     = {A new concept for the production of \(^{11}\)C-labelled radiotracers},
  series = {EJNMMI Radiopharmacy and Chemistry},
  volume    = {7},
  journal   = {EJNMMI Radiopharmacy and Chemistry},
  doi       = {10.1186/s41181-022-00159-y},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-300731},
  year      = {2022},
  abstract  = {Background The GMP-compliant production of radiopharmaceuticals has been performed using disposable units (cassettes) with a dedicated synthesis module. To expand this "plug 'n' synthesize" principle to a broader scope of modules we developed a pressure controlled setup that offers an alternative to the usual stepper motor controlled rotary valves. The new concept was successfully applied to the synthesis of N-methyl-[\(^{11}\)C]choline, L-S-methyl-[\(^{11}\)C]methionine and [11C]acetate. Results The target gas purification of cyclotron produced [\(^{11}\)C]CO\(_2\) and subsequent conversion to [\(^{11}\)C]MeI was carried out on a TRACERlab Fx C Pro module. The labelling reactions were controlled with a TRACERlab Fx FE module. With the presented modular principle we were able to produce N-methyl-[\(^{11}\)C]choline and L-S-methyl-[\(^{11}\)C]methionine by loading a reaction loop with neat N,N'-dimethylaminoethanol (DMAE) or an ethanol/water mixture of NaOH and L-homocysteine (L-HC), respectively and a subsequent reaction with [\(^{11}\)C]MeI. After 18 min N-methyl-[\(^{11}\)C]choline was isolated with 52\% decay corrected yield and a radiochemical purity of > 99\%. For L-S-methyl-[\(^{11}\)C]methionine the total reaction time was 19 min reaction, yielding 25\% of pure product (> 97\%). The reactor design was used as an exemplary model for the technically challenging [\(^{11}\)C]acetate synthesis. The disposable unit was filled with 1 mL MeMgCl (0.75 M) in tetrahydrofuran (THF) bevore [\(^{11}\)C]CO\(_2\) was passed through. After complete release of [\(^{11}\)C]CO\(_2\) the reaction mixture was quenched with water and guided through a series of ion exchangers (H\(^+\), Ag\(^+\) and OH\(^-\)). The product was retained on a strong anion exchanger, washed with water and finally extracted with saline. The product mixture was acidified and degassed to separate excess [\(^{11}\)C]CO\(_2\) before dispensing. Under these conditions the total reaction time was 18 ± 2 min and pure [\(^{11}\)C]acetate (n = 10) was isolated with a decay corrected yield of 51 ± 5\%. Conclusion Herein, we described a novel single use unit for the synthesis of carbon-11 labelled tracers for preclinical and clinical applications of N-methyl-[\(^{11}\)C]choline, L-S-methyl-[\(^{11}\)C]methionine and [11C]acetate.},
  language  = {en}
}