HI-NESS:a family of genetically encoded DNA labels based on a bacterial nucleoid-associated protein

The interplay between three-dimensional chromosome organisation and genomic processes such as replication and transcription necessitates in vivo studies of chromosome dynamics. Fluorescent organic dyes are often used for chromosome labelling in vivo. The mode of binding of these dyes to DNA cause its distortion, elongation, and partial unwinding. The structural changes induce DNA damage and interfere with the binding dynamics of chromatin-associated proteins, consequently perturbing gene expression, genome replication, and cell cycle progression. We have developed a minimally-perturbing, genetically encoded fluorescent DNA label consisting of a (photo-switchable) fluorescent protein fused to the DNA-binding domain of H-NS - a bacterial nucleoid-associated protein. We show that this DNA label, abbreviated as HI-NESS (H-NS-based indicator for nucleic acid stainings), is minimally-perturbing to genomic processes and labels chromosomes in eukaryotic cells in culture, and in zebrafish embryos with preferential binding to AT-rich chromatin.Genome Instability and Cance

Handmade clay bricks: chemical, physical and mechanical properties

The clay brick masonry that is much used in historical structures often is in a rather poor state of conservation. In order to intervene correctly in these buildings, it is convenient to characterize the old material. For this purpose, a large sample of clay brick specimens from the 12th to 19th century were collected from six Portuguese monasteries, and were characterized chemically, physically and mechanically. A large variability of the properties was found. Additionally, a sample of handmade new bricks, which are commonly used as replacing material, was also analysed. The results were compared to the old bricks and could be possibly adequate as substitution bricks. Still, significant differences were found in chemical composition, and in water absorption and porosity, which are much lower in modern handmade bricks. With respect to mechanical properties, the range of values found in old bricks was rather high and the degree of deterioration exhibited a large scatter, meaning that a conclusion is hardly possible.The authors gratefully acknowledge the Instituto de Gestao do Patrimonio Arquitectonico e Arqueologico (IGESPAR) for providing the old clay bricks used in the present work. The first author acknowledges the partial funding of this work by the FCT through the following scholarships POCTI SFRH/BD/6409/2001 and POCTI SFRH/BPD/26706/2005

HI-NESS: a family of genetically encoded DNA labels based on a bacterial nucleoid-associated protein

The interplay between three-dimensional chromosome organisation and genomic processes such as replication and transcription necessitates in vivo studies of chromosome dynamics. Fluorescent organic dyes are often used for chromosome labelling in vivo. The mode of binding of these dyes to DNA cause its distortion, elongation, and partial unwinding. The structural changes induce DNA damage and interfere with the binding dynamics of chromatin-associated proteins, consequently perturbing gene expression, genome replication, and cell cycle progression. We have developed a minimally-perturbing, genetically encoded fluorescent DNA label consisting of a (photo-switchable) fluorescent protein fused to the DNA-binding domain of H-NS - a bacterial nucleoid-associated protein. We show that this DNA label, abbreviated as HI-NESS (H-NS-based indicator for nucleic acid stainings), is minimally-perturbing to genomic processes and labels chromosomes in eukaryotic cells in culture, and in zebrafish embryos with preferential binding to AT-rich chromatin.Animal science