Engineered Orthogonal Quorum Sensing Systems for Synthetic Gene Regulation in Escherichia coli

Gene regulators that are controlled by membrane-permeable compounds called homoserine lactones (HSLs) have become popular tools for building synthetic gene networks that coordinate behaviors across populations of engineered bacteria. Synthetic HSL-signaling systems are derived from natural DNA and protein elements from microbial quorum signaling pathways. Crosstalk, where a single HSL can activate multiple regulators, can lead to faults in networks composed of parallel signaling pathways. Here, we report an investigation of quorum sensing components to identify synthetic pathways that exhibit little to no crosstalk in liquid and solid cultures. In previous work, we characterized the response of a single regulator (LuxR) to 10 distinct HSL-synthase enzymes. Our current study determined the responses of five different regulators (LuxR, LasR, TraR, BjaR, and AubR) to the same set of synthases. We identified two sets of orthogonal synthase-regulator pairs (BjaI/BjaR + EsaI/TraR and LasI/LasR + EsaI/TraR) that show little to no crosstalk when they are expressed in Escherichia coli BL21. These results expand the toolbox of characterized components for engineering microbial communities

Congenital Zika Virus Infection in Immunocompetent Mice Causes Postnatal Growth Impediment and Neurobehavioral Deficits

A small percentage of babies born to Zika virus (ZIKV)-infected mothers manifest severe defects at birth, including microcephaly. Among those who appeared healthy at birth, there are increasing reports of postnatal growth or developmental defects. However, the impact of congenital ZIKV infection in postnatal development is poorly understood. Here, we report that a mild congenital ZIKV-infection in pups born to immunocompetent pregnant mice did not display apparent defects at birth, but manifested postnatal growth impediments and neurobehavioral deficits, which include reduced locomotor and cognitive deficits that persisted into adulthood. We found that the brains of these pups were smaller, had a thinner cortical layer 1, displayed increased astrogliosis, decreased expression of microcephaly- and neuron development- related genes, and increased pathology as compared to mock-infected controls. In summary, our results showed that even a mild congenital ZIKV infection in immunocompetent mice could lead to postnatal deficits, providing definitive experimental evidence for a necessity to closely monitor postnatal growth and development of presumably healthy human infants, whose mothers were exposed to ZIKV infection during pregnancy

Manual / Issue 9 / Out of Line

Manual, a journal about art and its making. Out of Line. The nineth issue. This issue of *Manual*—themed Out of Line—is a collection about the way that lines disrupt, point outward. In poetry, the attention to detail one takes in crafting a line is all about making the line disappear, making something it holds to take front stage. . . . The space between the lines creating the image . . . the space around that argues for the importance of all that the lines hold. Manual 9 (Out of Line) complemented Lines of Thought: Drawing from Michelangelo to Now, presented in collaboration with the British Museum, on view at the RISD Museum October 6, 2017 to January 7, 2018. Softcover, 76 pages. Published 2017 by the RISD Museum. Manual 9 (Out of Line) contributors include Fida Adely, Reginald Dwayne Betts, Stefano Bloch, Mimi Cabell, Namita Vijay Dharia, Douglas W. Doe, Jared A. Goldstein, Lucinda Hitchcock, Jan Howard, Kate Irvin, Douglas Kearney, Amber Lopez, Jeffrey Moser, Sheida Soleimani, and Craig Taylor.https://digitalcommons.risd.edu/risdmuseum_journals/1035/thumbnail.jp

Representativeness of the European social partner organisations : human health sector

Aquesta publicació s'elabora a partir de les contribucions de cadascú dels membres nacionals que integren la Network of Eufound Correspondent. Pel cas d'Espanya la contribució ha estat realitzada per l'Oscar MolinaThis study provides information allowing for an assessment of the representativeness of the actors involved in the European sectoral social dialogue committee for the human health sector. Their relative representativeness legitimises their right to be consulted, their role and effective participation in the European sectoral social dialogue, and their capacity to negotiate agreements. The aim of Eurofound's studies on representativeness is to identify the relevant national and European social partner organisations in the field of industrial relations in the EU Member States. This study identified the European Federation of Public Service Unions (EPSU) - representing employees - and the European Hospital and Healthcare Employers' Association (HOSPEEM) - representing employers - as the most representative European-level social partner organisations in the human health sector. The member organisations of the European Confederation of Independent Trade Unions (CESI) and UNI Europa also organise employees in the sector in several Member States

Diagnosis of sheep fasciolosis caused by Fasciola hepatica using cathepsin L enzyme-linked immunosorbent assays (ELISA)

Publication history: Accepted - 3 July 2021; Published online - 6 July 2021.Fasciolosis, a global parasitic disease of agricultural livestock, is caused by the liver fluke Fasciola hepatica. Management and strategic control of fasciolosis on farms depends on early assessment of the extent of disease so that control measures can be implemented quickly. Traditionally, this has relied on the detection of eggs in the faeces of animals, a laborious method that lacks sensitivity, especially for sub-clinical infections, and identifies chronic infections only. Enzyme linked immunosorbent assays (ELISA) offer a quicker and more sensitive serological means of diagnosis that could detect early acute infection before significant liver damage occurs. The performance of three functionally-active recombinant forms of the major F. hepatica secreted cathepsins L, rFhCL1, rFhCL2, rFhCL3, and a cathepsin B, rFhCB3, were evaluated as antigens in an indirect ELISA to serologically diagnose liver fluke infection in experimentally and naturally infected sheep. rFhCL1 and rFhCL3 were the most effective of the four antigens detecting fasciolosis in sheep as early as three weeks after experimental infection, at least five weeks earlier than both coproantigen and faecal egg tests. In addition, the rFhCL1 and rFhCL3 ELISAs had a very low detection limit for liver fluke in lambs exposed to natural infection on pastures and thus could play a major role in the surveillance of farms and a ‘test and treat’ approach to disease management. Finally, antibodies to all three cathepsin L proteases remain high throughout chronic infection but decline rapidly after drug treatment with the flukicide, triclabendazole, implying that the test may be adapted to trace the effectiveness of drug treatment.This work was supported by a European Research Council Advanced Grant (HELIVAC, 322725) and Science Foundation Ireland (SFI) Professorship grant (17/RP/5368) awarded to J.P. Dalton

Effects of antiplatelet therapy on stroke risk by brain imaging features of intracerebral haemorrhage and cerebral small vessel diseases: subgroup analyses of the RESTART randomised, open-label trial

Background Findings from the RESTART trial suggest that starting antiplatelet therapy might reduce the risk of recurrent symptomatic intracerebral haemorrhage compared with avoiding antiplatelet therapy. Brain imaging features of intracerebral haemorrhage and cerebral small vessel diseases (such as cerebral microbleeds) are associated with greater risks of recurrent intracerebral haemorrhage. We did subgroup analyses of the RESTART trial to explore whether these brain imaging features modify the effects of antiplatelet therapy

Leptin-signaling inhibition results in efficient anti-tumor activity in estrogen receptor positive or negative breast cancer

Introduction: We have shown previously that treatment with pegylated leptin peptide receptor antagonist 2 (PEG-LPrA2) reduced the expression of vascular endothelial growth factor (VEGF), vascular endothelial growth factor receptor type 2 (VEGFR2) and growth of 4T1-breast cancer (BC) in syngeneic mice. In this investigation, PEG-LPrA2 was used to evaluate whether the inhibition of leptin signaling has differential impact on the expression of pro-angiogenic and pro-proliferative molecules and growth of human estrogen receptor-positive (ER+) and estrogen receptor-negative (ER-) BC xenografts hosted by immunodeficient mice. Methods: To test the contribution of leptin signaling to BC growth and expression of leptin-targeted molecules, PEG-LPrA2 treatment was applied to severe immunodeficient mice hosting established ER+ (MCF-7 cells; ovariectomized/supplemented with estradiol) and ER- (MDA-MB231 cells) BC xenografts. To further assess leptin and PEG-LPrA2 effects on ER+ and ER- BC, the expression of VEGF and VEGFR2 (protein and mRNA) was investigated in cell cultures. Results: PEG-LPrA2 more effectively reduced the growth of ER+ (>40-fold) than ER- BC (twofold) and expression of pro-angiogenic (VEGF/VEGFR2, leptin/leptin receptor OB-R, and IL-1 receptor type I) and pro-proliferative molecules (proliferating cell nuclear antigen and cyclin D1) in ER+ than in ER- BC. Mouse tumor stroma in ER+ BC expressed high levels of VEGF and leptin that was induced by leptin signaling. Leptin upregulated the transcriptional expression of VEGF/VEGFR2 in MCF-7 and MDA-MB231 cells. Conclusions: These results suggest that leptin signaling plays an important role in the growth of both ER+ and ER- BC that is associated with the leptin regulation of pro-angiogenic and pro-proliferative molecules. These data provide support for the potential use of leptin-signaling inhibition as a novel treatment for ER+ and ER- BC

Molten salt synthesis of Ce doped zirconolite for the immobilisation of pyroprocessing wastes and separated plutonium

Molten salt mediated synthesis of zirconolite Ca0.9Zr0.9Ce0.2Ti2O7 was investigated, as a target ceramic matrix for the clean-up of waste molten salts from pyroprocessing of spent nuclear fuels and the immobilisation of separated plutonium. A systematic study of reaction variables, including, reaction temperature, time, atmosphere, reagents and composition, was made to optimise the yield of the target zirconolite phase. Zirconolite 2 M and 3T polytypes were formed as the major phase (with minor perovskite) between 1000 – 1400 °C, in air, with the relative proportion of 2 M polytype increasing with temperature. Synthesis under 5% H2/N2 or Ar increased the proportion of minor perovskite phase and reduced the yield of the zirconolite phase. The yield of zirconolite polytypes was maximised with the addition of 10 wt% TiO2 and 5 wt% TiO2, yielding 91.7 ± 2.0 wt% zirconolite, primarily as the 2 M polytype, after reaction at 1200 °C for 2 h, in air. The particle size and morphology of the zirconolite product bears a close resemblance to that of the TiO2 precursor, demonstrating a dominant template growth mechanism. Although the molten salt mediated synthesis of zirconolite is effective at lower reaction temperature and time, compared to reactive sintering, this investigation has demonstrated that the approach does not offer any clear advantage with over conventional reactive sintering for the envisaged application