Benchmarking of 4C-seq pipelines based on real and simulated data

"jats:sec" "jats:title"Motivation"/jats:title" "jats:p"With its capacity for high-resolution data output in one region of interest, chromosome conformation capture combined with high-throughput sequencing (4C-seq) is a state-of-the-art next-generation sequencing technique that provides epigenetic insights, and regularly advances current medical research. However, 4C-seq data are complex and prone to biases, and while specialized programs exist, an unbiased, extensive benchmarking is still lacking. Furthermore, neither substantial datasets with fully characterized ground truth, nor simulation programs for realistic 4C-seq data have been published."/jats:p" "/jats:sec" "jats:sec" "jats:title"Results"/jats:title" "jats:p"We conducted a benchmarking study on 66 4C-seq samples from 20 datasets, and developed a novel 4C-seq simulation software, Basic4CSim, to allow for detailed comparisons of 4C-seq algorithms on 50 simulated datasets with 10–120 samples each. Simulations and benchmarking were adapted to address different characteristics of 4C-seq data. Simulated data were compared with published samples to validate simulation settings. We identified differences between 4C-seq algorithms in terms of precision, recall, interaction structure, and run time, and observed general trends. Novel differential pipeline versions of single-sample based 4C-seq algorithms were included in the benchmarking. While no single tool was optimally suited for both near-cis and far-cis, and both single-sample and differential analyses, choosing a high-performing algorithm variant did improve results considerably. For near-cis scenarios, r3Cseq, peakC and FourCSeq offered high precision, while fourSig demonstrated high overall F1 scores in far-cis analyses. Finally, 4C-seq simulations may aid in the development of improved analysis algorithms."/jats:p" "/jats:sec" "jats:sec" "jats:title"Availability and implementation"/jats:title" "jats:p"Basic4CSim is available at https://github.com/walter–ca/Basic4CSim."/jats:p" "/jats:sec" "jats:sec" "jats:title"Supplementary information"/jats:title" "jats:p"Supplementary data are available at Bioinformatics online."/jats:p" "/jats:sec Document type: Articl

C-axis optical properties of high Tc cuprates

A review is given of the experimental status of the interlayer coupling energy in the cuprates. A second c-axis plasmon is identified in the double layer compound Y123 for various dopings. The anomalous transport properties along the c-direction and in the planar directions are compared to model calculations based on strongly anisotropic scattering. An excellent description of the optical data at optimal doping is obtained if an anomalously large anisotropy of the scattering rate between cold spots and hot spots is assumed. This raises questions as to the physical meaning of these parameters.Comment: 4 pages, LaTeX, espcrc2.sty, 3 figures in encapsulated postscript forma

Label-free enrichment of avian Leucocytozoon using flow cytrometric sorting

Chakarov N, Greiner J, Hauser S, et al. Label-free enrichment of avian Leucocytozoon using flow cytrometric sorting. Parasitology. 2012;139(12):1547-1552

Efficient animal-serum free 3D cultivation method for adult human neural crest-derived stem cell therapeutics

Greiner J, Hauser S, Widera D, et al. Efficient animal-serum free 3D cultivation method for adult human neural crest-derived stem cell therapeutics. European Cells & Materials. 2011;22:403-419

Efficient animal-serum free 3D cultivation method for adult human neural crest-derived stem cell therapeutics

Due to their broad differentiation potential and their persistence into adulthood, human neural crest-derived stem cells (NCSCs) harbour great potential for autologous cellular therapies, which include the treatment of neurodegenerative diseases and replacement of complex tissues containing various cell types, as in the case of musculoskeletal injuries. The use of serum-free approaches often results in insufficient proliferation of stem cells and foetal calf serum implicates the use of xenogenic medium components. Thus, there is much need for alternative cultivation strategies. In this study we describe for the first time a novel, human blood plasma based semi-solid medium for cultivation of human NCSCs. We cultivated human neural crest-derived inferior turbinate stem cells (ITSCs) within a blood plasma matrix, where they revealed higher proliferation rates compared to a standard serum-free approach. Three-dimensionality of the matrix was investigated using helium ion microscopy. ITSCs grew within the matrix as revealed by laser scanning microscopy. Genetic stability and maintenance of stemness characteristics were assured in 3D cultivated ITSCs, as demonstrated by unchanged expression profile and the capability for self-renewal. ITSCs pre-cultivated in the 3D matrix differentiated efficiently into ectodermal and mesodermal cell types, particularly including osteogenic cell types. Furthermore, ITSCs cultivated as described here could be easily infected with lentiviruses directly in substrate for potential tracing or gene therapeutic approaches. Taken together, the use of human blood plasma as an additive for a completely defined medium points towards a personalisable and autologous cultivation of human neural crest-derived stem cells under clinical grade conditions

Temporal autoregulation during human PU.1 locus SubTAD formation

Epigenetic control of gene expression occurs within discrete spatial chromosomal units called topologically associating domains (TADs), but the exact spatial requirements of most genes are unknown; this is of particular interest for genes involved in cancer. We therefore applied high-resolution chromosomal conformation capture sequencing to map the three-dimensional (3D) organization of the human locus encoding the key myeloid transcription factor PU.1 in healthy monocytes and acute myeloid leukemia (AML) cells. We identified a dynamic similar to 75-kb unit (SubTAD) as the genomic region in which spatial interactions between PU.1 gene regulatory elements occur during myeloid differentiation and are interrupted in AML. Within this SubTAD, proper initiation of the spatial chromosomal interactions requires PU.1 autoregulation and recruitment of the chromatin-adaptor protein LDB1 (LIM domain-binding protein 1). However, once these spatial interactions have occurred, LDB1 stabilizes them independently of PU.1 autoregulation. Thus, our data support that PU.1 autoregulates its expression in a "hit-and-run" manner by initiating stable chromosomal loops that result in a transcriptionally active chromatin architecture