Strain construction to test the significance of antiactivation in Pseudomonas quorum sensing

The quorum sensing regulatory pathway has been extensively studied for its impact on the virulence of Pseudomonas aeruginosa, an opportunistic pathogen that causes acute and chronic infections in immunocompromised individuals such as those suffering from the genetic disorder cystic fibrosis. Part of the quorum sensing pathway that has not been adequately described is the mechanism behind the accumulation of a threshold concentration of autoinducer. Recently, a protein, QteE, has been shown to inhibit the transcriptional regulator LasR. LasR binds the autoinducer molecule 3-oxo-dodecanoyl-homoserine lactone produced by the synthase LasI. QteE could also have a role in preventing the autoinducer of one cell binding its own LasR and inducing quorum sensing without a threshold concentration of autoinducer, essentially “short-circuiting” the cell. A co-culturing experiment with two QteE-deficient strains of P. aeruginosa, one LasI-proficient and the other LasI-deficient, would determine if this hypothesis is correct. The co-culturing experiment requires a method to differentiate between two different strains of bacteria grown together in the same medium, which in this case is tagging one strain with a red fluorescent protein, mCherry. For this thesis, I integrated a plasmid containing mCherry (pVM3) into the chromosome of the P. aeruginosa strains, and then confirmed their fluorescence with spectroscopy and microscopy. I also attempted to confirm the chromosomal location of the insertion by PCR

The CUL4-DDB1 ubiquitin ligase complex controls adult and embryonic stem cell differentiation and homeostasis

Little is known on post-transcriptional regulation of adult and embryonic stem cell maintenance and differentiation. Here we characterize the role of Ddb1, a component of the CUL4-DDB1 ubiquitin ligase complex. Ddb1 is highly expressed in multipotent hematopoietic progenitors and its deletion leads to abrogation of both adult and fetal hematopoiesis, targeting specifically transiently amplifying progenitor subsets. However, Ddb1 deletion in non-dividing lymphocytes has no discernible phenotypes. Ddb1 silencing activates Trp53 pathway and leads to significant effects on cell cycle progression and rapid apoptosis. The abrogation of hematopoietic progenitor cells can be partially rescued by simultaneous deletion of Trp53. Conversely, depletion of DDB1 in embryonic stem cell (ESC) leads to differentiation albeit negative effects on cell cycle and apoptosis. Mass spectrometry reveals differing protein interactions between DDB1 and distinct DCAFs, the substrate recognizing components of the E3 complex, between cell types. Our studies identify CUL4-DDB1 complex as a novel post-translational regulator of stem and progenitor maintenance and differentiation

Spin correlations and exchange in square lattice frustrated ferromagnets

The J1-J2 model on a square lattice exhibits a rich variety of different forms of magnetic order that depend sensitively on the ratio of exchange constants J2/J1. We use bulk magnetometry and polarized neutron scattering to determine J1 and J2 unambiguously for two materials in a new family of vanadium phosphates, Pb2VO(PO4)2 and SrZnVO(PO4)2, and we find that they have ferromagnetic J1. The ordered moment in the collinear antiferromagnetic ground state is reduced, and the diffuse magnetic scattering is enhanced, as the predicted bond-nematic region of the phase diagram is approached.Comment: 4 pages, 4 figure

Thallium isotopes in early diagenetic pyrite – a paleoredox proxy?

Author Posting. © The Author(s), 2011. This is the author's version of the work. It is posted here by permission of Elsevier B.V. for personal use, not for redistribution. The definitive version was published in Geochimica et Cosmochimica Acta 75 (2011): 6690-6704, doi:10.1016/j.gca.2011.07.047.This paper presents the first study of Tl isotopes in early diagenetic pyrite. Measurements from two sections deposited during the Toarcian Ocean Anoxic Event (T-OAE, ~183Ma) are compared with data from Late Neogene (<10Ma) pyrite samples from ODP legs 165 and 167 that were deposited in relatively oxic marine environments. The Tl isotope compositions of Late Neogene pyrites are all significantly heavier than seawater, which most likely indicates that Tl in diagenetic pyrite is partially sourced from ferromanganese oxy-hydroxides that are known to display relatively heavy Tl isotope signatures. One of the T-OAE sections from Peniche in Portugal displays pyrite thallium isotope compositions indistinguishable from Late Neogene samples, whereas samples from Yorkshire in the UK are depleted in the heavy isotope of Tl. These lighter compositions are best explained by the lack of ferromanganese precipitation at the sediment–water interface due the sulphidic (euxinic) conditions thought to be prevalent in the Cleveland Basin where the Yorkshire section was deposited. The heavier signatures in the Peniche samples appear to result from an oxic water column that enabled precipitation of ferromanganese oxy-hydroxides at the sediment–water interface. The Tl isotope profile from Yorkshire is also compared with previously published molybdenum isotope ratios determined on the same sedimentary succession. There is a suggestion of an anti-correlation between these two isotope systems, which is consistent with the expected isotope shifts that occur in seawater when marine oxic (ferromanganese minerals) fluxes fluctuate. The results outlined here represent the first evidence that Tl isotopes in early diagenetic pyrite have potential to reveal variations in past ocean oxygenation on a local scale and potentially also for global oceans. However, much more information about Tl isotopes in different marine environments, especially in anoxic/euxinic basins, is needed before Tl isotopes can be confidently utilized as a paleo-redox tracer.SGN is funded by a NERC fellowship

Variability in chemotherapy delivery for elderly women with advanced stage ovarian cancer and its impact on survival

Given the survival benefits of adjuvant chemotherapy for advanced ovarian cancer (OC), we examined the associations of survival with the time interval from debulking surgery to initiation of chemotherapy and with the duration of chemotherapy. Among patients ⩾65 years with stages III/IV OC diagnosed between 1991 and 2002 in the Surveillance, Epidemiology, and End Results-Medicare database, we developed regression models of predictors of the time interval from surgery to initiation of chemotherapy and of the total duration of chemotherapy. Survival was examined with Cox proportional hazards models. Among 2558 patients, 1712 (67%) initiated chemotherapy within 6 weeks of debulking surgery, while 846 (33%) began treatment >6 weeks. Older age, black race, being unmarried, and increased comorbidities were associated with delayed initiation of chemotherapy. Delay of chemotherapy was associated with an increase in mortality (hazard ratio (HR)=1.11; 95% CI, 1.0–1.2). Among 1932 patients in the duration of treatment analysis, the 1218 (63%) treated for 3–7 months had better survival than the 714 (37%) treated for ⩽3 months (HR=0.84; 95% CI, 0.75–0.94). This analysis represents one of the few studies describing treatment delivery and outcome in women with advanced OC. Delayed initiation and early discontinuation of chemotherapy were common and associated with increased mortality

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

Assembly and architecture of precursor nodes during fission yeast cytokinesis

Mapping of fission yeast precursor node interaction modules and assembly reveals important steps in contractile ring assembly