The preservation of morphological features, such as protrusions and concavities, and of the topology of input shapes is important when establishing reference data for benchmarking segmentation algorithms or when constructing a mean or median shape. We present a contourwise topology-preserving fusion method, called shape-aware topology-preserving means (SATM), for merging complex simply connected shapes. The method is based on key point matching and piecewise contour averaging. Unlike existing pixelwise and contourwise fusion methods, SATM preserves topology and does not smooth morphological features. We also present a detailed comparison of SATM with state-of-the-art fusion techniques for the purpose of benchmarking and median shape construction. Our experiments show that SATM outperforms these techniques in terms of shape-related measures that reflect shape complexity, manifesting itself as a reliable method for both establishing a consensus of segmentation annotations and for computing mean shapes.

• Keywords
• Average shape, Mean shape, Median shape, Segmentation mask fusion, Shape analysis,
• Publication type
• Journal Article MeSH

The nucleolus is a well-organized site of ribosomal gene transcription. Moreover, many DNA repair pathway proteins, including ATM, ATR kinases, MRE11, PARP1 and Ku70/80, localize to the nucleolus (Moore et al., 2011 ). We analyzed the consequences of DNA damage in nucleoli following ultraviolet A (UVA), C (UVC), or γ-irradiation in order to test whether and how radiation-mediated genome injury affects local motion and morphology of nucleoli. Because exposure to radiation sources can induce changes in the pattern of UBF1-positive nucleolar regions, we visualized nucleoli in living cells by GFP-UBF1 expression for subsequent morphological analyses and local motion studies. UVA radiation, but not 5 Gy of γ-rays, induced apoptosis as analyzed by an advanced computational method. In non-apoptotic cells, we observed that γ-radiation caused nucleolar re-positioning over time and changed several morphological parameters, including the size of the nucleolus and the area of individual UBF1-positive foci. Radiation-induced nucleoli re-arrangement was observed particularly in G2 phase of the cell cycle, indicating repair of ribosomal genes in G2 phase and implying that nucleoli are less stable, thus sensitive to radiation, in G2 phase.

• Keywords
• DNA damage, UBF1, live cells, nucleolus, nuncleoli tracking,
• MeSH
• Apoptosis radiation effects   MeSH
• Cell Nucleolus radiation effects   MeSH
• Cell Line MeSH
• Cell Cycle radiation effects   MeSH
• G2 Phase radiation effects   MeSH
• Transcription, Genetic MeSH
• Mice MeSH
• Cell Line, Tumor MeSH
• DNA Damage radiation effects   MeSH
• Pol1 Transcription Initiation Complex Proteins genetics   metabolism   MeSH
• Ultraviolet Rays MeSH
• Computational Biology MeSH
• Gamma Rays adverse effects   MeSH
• Animals MeSH
• Check Tag
• Mice MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• transcription factor UBF MeSH Browser
• Pol1 Transcription Initiation Complex Proteins MeSH

BACKGROUND: Protein exchange kinetics correlate with the level of chromatin condensation and, in many cases, with the level of transcription. We used fluorescence recovery after photobleaching (FRAP) to analyse the kinetics of 18 proteins and determine the relationships between nuclear arrangement, protein molecular weight, global transcription level, and recovery kinetics. In particular, we studied heterochromatin-specific heterochromatin protein 1β (HP1β) B lymphoma Mo-MLV insertion region 1 (BMI1), and telomeric-repeat binding factor 1 (TRF1) proteins, and nucleolus-related proteins, upstream binding factor (UBF) and RNA polymerase I large subunit (RPA194). We considered whether the trajectories and kinetics of particular proteins change in response to histone hyperacetylation by histone deacetylase (HDAC) inhibitors or after suppression of transcription by actinomycin D. RESULTS: We show that protein dynamics are influenced by many factors and events, including nuclear pattern and transcription activity. A slower recovery after photobleaching was found when proteins, such as HP1β, BMI1, TRF1, and others accumulated at specific foci. In identical cells, proteins that were evenly dispersed throughout the nucleoplasm recovered more rapidly. Distinct trajectories for HP1β, BMI1, and TRF1 were observed after hyperacetylation or suppression of transcription. The relationship between protein trajectory and transcription level was confirmed for telomeric protein TRF1, but not for HP1β or BMI1 proteins. Moreover, heterogeneity of foci movement was especially observed when we made distinctions between centrally and peripherally positioned foci. CONCLUSION: Based on our results, we propose that protein kinetics are likely influenced by several factors, including chromatin condensation, differentiation, local protein density, protein binding efficiency, and nuclear pattern. These factors and events likely cooperate to dictate the mobility of particular proteins.

• Publication type
• Journal Article MeSH