Bright ligand-activatable fluorescent protein for high-quality multicolor live-cell super-resolution microscopy

We introduce UnaG as a green-to-dark photoswitching fluorescent protein capable of high-quality super-resolution imaging with photon numbers equivalent to the brightest photoswitchable red protein. UnaG only fluoresces upon binding of a fluorogenic metabolite, bilirubin, enabling UV-free reversible photoswitching with easily controllable kinetics and low background under Epi illumination. The on- and off-switching rates are controlled by the concentration of the ligand and the excitation light intensity, respectively, where the dissolved oxygen also promotes the off-switching. The photo-oxidation reaction mechanism of bilirubin in UnaG suggests that the lack of ligand-protein covalent bond allows the oxidized ligand to detach from the protein, emptying the binding cavity for rebinding to a fresh ligand molecule. We demonstrate super-resolution single-molecule localization imaging of various subcellular structures genetically encoded with UnaG, which enables facile labeling and simultaneous multicolor imaging of live cells. UnaG has the promise of becoming a default protein for high-performance super-resolution imaging. Photoconvertible proteins occupy two color channels thereby limiting multicolour localisation microscopy applications. Here the authors present UnaG, a new green-to-dark photoswitching fluorescent protein for super-resolution imaging, whose activation is based on a noncovalent binding with bilirubin

Parâmetros de cultivo e a enfermidade da mancha-branca em fazendas de camarões de Santa Catarina

O objetivo deste trabalho foi avaliar a relação entre parâmetros físico-químicos e biológicos do cultivo de camarão com a enfermidade causada pelo vírus da síndrome da mancha-branca (WSSV). Foram avaliadas a mortalidade, as características físicas e químicas da água e do solo, e foram realizados os exames e as análises microscópicas a fresco, presença de víbrios na hemolinfa, além do diagnóstico histopatológico e molecular (PCR) do WSSV, em viveiros de oito fazendas de Santa Catarina, entre 2008 e 2009. O vírus foi detectado em cinco fazendas com registros da enfermidade em ciclos anteriores. A temperatura da água foi semelhante entre as fazendas com e sem WSSV, e a mortalidade ocorreu a intervalos ascendentes entre 24,6 e 29,3ºC. Não foram encontradas diferenças significativas nos parâmetros de análises microscópicas a fresco, tempo de coagulação e presença de víbrios na hemolinfa e sinais clínicos, entre viveiros com e sem WSSV. As concentrações de nitrito, sílica, fenol e alcalinidade na água e pH do solo apresentaram diferenças significativas entre os viveiros afetados ou não por WSSV. Os valores de nitrito, sílica e pH do solo estavam dentro dos limites recomendados para o cultivo de camarões, no entanto, isto não ocorreu com a alcalinidade e a concentração de fenol, o que sugere uma relação dos últimos com a manifestação da enfermidade da mancha-branca

The role of tenascin-C in tissue injury and tumorigenesis

The extracellular matrix molecule tenascin-C is highly expressed during embryonic development, tissue repair and in pathological situations such as chronic inflammation and cancer. Tenascin-C interacts with several other extracellular matrix molecules and cell-surface receptors, thus affecting tissue architecture, tissue resilience and cell responses. Tenascin-C modulates cell migration, proliferation and cellular signaling through induction of pro-inflammatory cytokines and oncogenic signaling molecules amongst other mechanisms. Given the causal role of inflammation in cancer progression, common mechanisms might be controlled by tenascin-C during both events. Drugs targeting the expression or function of tenascin-C or the tenascin-C protein itself are currently being developed and some drugs have already reached advanced clinical trials. This generates hope that increased knowledge about tenascin-C will further improve management of diseases with high tenascin-C expression such as chronic inflammation, heart failure, artheriosclerosis and cancer

Comparative genomic analysis of innate immunity reveals novel and conserved components in crustacean food crop species

Abstract Background Growing global demands for crustacean food crop species have driven large investments in aquaculture research worldwide. However, large-scale production is susceptible to pathogen-mediated destruction particularly in developing economies. Thus, a thorough understanding of the immune system components of food crop species is imperative for research to combat pathogens. Results Through a comparative genomics approach utilising extant data from 55 species, we describe the innate immune system of the class Malacostraca, which includes all food crop species. We identify 7407 malacostracan genes from 39 gene families implicated in different aspects of host defence and demonstrate dynamic evolution of innate immunity components within this group. Malacostracans have achieved flexibility in recognising infectious agents through divergent evolution and expansion of pathogen recognition receptors genes. Antiviral RNAi, Toll and JAK-STAT signal transduction pathways have remained conserved within Malacostraca, although the Imd pathway appears to lack several key components. Immune effectors such as the antimicrobial peptides (AMPs) have unique evolutionary profiles, with many malacostracan AMPs not found in other arthropods. Lastly, we describe four putative novel immune gene families, potentially representing important evolutionary novelties of the malacostracan immune system. Conclusion Our analyses across the broader Malacostraca have allowed us to not only draw analogies with other arthropods but also to identify evolutionary novelties in immune modulation components and form strong hypotheses as to when key pathways have evolved or diverged. This will serve as a key resource for future immunology research in crustacean food crops