Structural changes that occur upon photolysis of the Fe(II)a3–CO complex in the cytochrome ba3-oxidase of Thermus thermophilus: A combined X-ray crystallographic and infrared spectral study demonstrates CO binding to CuB

AbstractThe purpose of the work was to provide a crystallographic demonstration of the venerable idea that CO photolyzed from ferrous heme-a3 moves to the nearby cuprous ion in the cytochrome c oxidases. Crystal structures of CO-bound cytochrome ba3-oxidase from Thermus thermophilus, determined at ~2.8–3.2Å resolution, reveal a Fe–C distance of ~2.0Å, a Cu–O distance of 2.4Å and a Fe–C–O angle of ~126°. Upon photodissociation at 100K, X-ray structures indicate loss of Fea3–CO and appearance of CuB–CO having a Cu–C distance of ~1.9Å and an O–Fe distance of ~2.3Å. Absolute FTIR spectra recorded from single crystals of reduced ba3–CO that had not been exposed to X-ray radiation, showed several peaks around 1975cm−1; after photolysis at 100K, the absolute FTIR spectra also showed a significant peak at 2050cm−1. Analysis of the ‘light’ minus ‘dark’ difference spectra showed four very sharp CO stretching bands at 1970cm−1, 1977cm−1, 1981cm−1, and 1985cm−1, previously assigned to the Fea3–CO complex, and a significantly broader CO stretching band centered at ~2050cm−1, previously assigned to the CO stretching frequency of CuB bound CO. As expected for light propagating along the tetragonal axis of the P43212 space group, the single crystal spectra exhibit negligible dichroism. Absolute FTIR spectrometry of a CO-laden ba3 crystal, exposed to an amount of X-ray radiation required to obtain structural data sets before FTIR characterization, showed a significant signal due to photogenerated CO2 at 2337cm−1 and one from traces of CO at 2133cm−1; while bands associated with CO bound to either Fea3 or to CuB in “light” minus “dark” FTIR difference spectra shifted and broadened in response to X-ray exposure. In spite of considerable radiation damage to the crystals, both X-ray analysis at 2.8 and 3.2Å and FTIR spectra support the long-held position that photolysis of Fea3–CO in cytochrome c oxidases leads to significant trapping of the CO on the CuB atom; Fea3 and CuB ligation, at the resolutions reported here, are otherwise unaltered. This article is part of a Special Issue entitled: Respiratory Oxidases

Disparities in inflammation between non-Hispanic black and white individuals with lung cancer in the Greater Chicago Metropolitan area

BackgroundLung cancer incidence and mortality rates are higher in Non-Hispanic Black (NHB) compared to Non-Hispanic White (NHW) individuals in the Chicago metropolitan area, which may be related to exposure to chronic stress which may increase inflammation.Specific aimWe investigated disparities in inflammation as measured by neutrophil to lymphocyte ratio (NLR) in individuals with lung cancer by race and by neighborhood concentrated disadvantage index (CDI).MethodsThis retrospective, cross-sectional study included 263 NHB and NHW adults with lung cancer. We analyzed NLR as a continuous and categorical variable to determine degree and prevalence of inflammation. We used Mann Whitney U, t-tests, Chi square tests, linear and logistic regression models as appropriate.ResultsMore than 60% of subjects had inflammation (NLR ≥ 3) at lung cancer diagnosis. The degree of inflammation was significantly lower in NHB (NLR 5.50 +/- 7.45) compared to NHW individuals (NLR 6.53 +/- 6.53; p=0.01) but did not differ by neighborhood CDI. The prevalence of inflammation (NLR ≥ 3) was significantly lower in NHB (55.07%) compared to NHW individuals (71.20%; p<0.01) and in those from the most disadvantaged (54.07%) compared to the least disadvantaged (71.88%; p<0.01) neighborhoods.ConclusionAt lung cancer diagnosis, there is a lower degree and prevalence of inflammation in NHB compared to NHW individuals, and lower prevalence in those residing in the most disadvantaged neighborhoods. Further research is needed to determine mechanisms of inflammation that may be contributing to lung cancer disparities as well as whether NLR is an appropriate biomarker when examining racial differences in inflammation

Farnesoid X receptor and liver X receptor ligands initiate formation of coated platelets

The liver X receptors (LXRs) and farnesoid X receptor (FXR) have been identified in human platelets. Ligands of these receptors have been shown to have nongenomic inhibitory effects on platelet activation by platelet agonists. This, however, seems contradictory with the platelet hyper-reactivity that is associated with several pathological conditions that are associated with increased circulating levels of molecules that are LXR and FXR ligands, such as hyperlipidemia, type 2 diabetes mellitus, and obesity. We, therefore, investigated whether ligands for the LXR and FXR receptors were capable of priming platelets to the activated state without stimulation by platelet agonists. Treatment of platelets with ligands for LXR and FXR converted platelets to the procoagulant state, with increases in phosphatidylserine exposure, platelet swelling, reduced membrane integrity, depolarization of the mitochondrial membrane, and microparticle release observed. Additionally, platelets also displayed features associated with coated platelets such as P-selectin exposure, fibrinogen binding, fibrin generation that is supported by increased serine protease activity, and inhibition of integrin αIIbβ3. LXR and FXR ligand-induced formation of coated platelets was found to be dependent on both reactive oxygen species and intracellular calcium mobilization, and for FXR ligands, this process was found to be dependent on cyclophilin D. We conclude that treatment with LXR and FXR ligands initiates coated platelet formation, which is thought to support coagulation but results in desensitization to platelet stimuli through inhibition of αIIbβ3 consistent with their ability to inhibit platelet function and stable thrombus formation in vivo

COLD GASS, an IRAM legacy survey of molecular gas in massive galaxies: I. Relations between H2, HI, stellar content and structural properties

We are conducting COLD GASS, a legacy survey for molecular gas in nearby galaxies. Using the IRAM 30m telescope, we measure the CO(1-0) line in a sample of ~350 nearby (D=100-200 Mpc), massive galaxies (log(M*/Msun)>10.0). The sample is selected purely according to stellar mass, and therefore provides an unbiased view of molecular gas in these systems. By combining the IRAM data with SDSS photometry and spectroscopy, GALEX imaging and high-quality Arecibo HI data, we investigate the partition of condensed baryons between stars, atomic gas and molecular gas in 0.1-10L* galaxies. In this paper, we present CO luminosities and molecular hydrogen masses for the first 222 galaxies. The overall CO detection rate is 54%, but our survey also uncovers the existence of sharp thresholds in galaxy structural parameters such as stellar mass surface density and concentration index, below which all galaxies have a measurable cold gas component but above which the detection rate of the CO line drops suddenly. The mean molecular gas fraction MH2/M* of the CO detections is 0.066+/-0.039, and this fraction does not depend on stellar mass, but is a strong function of NUV-r colour. Through stacking, we set a firm upper limit of MH2/M*=0.0016+/-0.0005 for red galaxies with NUV-r>5.0. The average molecular-to-atomic hydrogen ratio in present-day galaxies is 0.3, with significant scatter from one galaxy to the next. The existence of strong detection thresholds in both the HI and CO lines suggests that "quenching" processes have occurred in these systems. Intriguingly, atomic gas strongly dominates in the minority of galaxies with significant cold gas that lie above these thresholds. This suggests that some re-accretion of gas may still be possible following the quenching event.Comment: Accepted for publications in MNRAS. 32 pages, 25 figure

Toward a 21st-century health care system: Recommendations for health care reform

The coverage, cost, and quality problems of the U.S. health care system are evident. Sustainable health care reform must go beyond financing expanded access to care to substantially changing the organization and delivery of care. The FRESH-Thinking Project (www.fresh-thinking.org) held a series of workshops during which physicians, health policy experts, health insurance executives, business leaders, hospital administrators, economists, and others who represent diverse perspectives came together. This group agreed that the following 8 recommendations are fundamental to successful reform: 1. Replace the current fee-for-service payment system with a payment system that encourages and rewards innovation in the efficient delivery of quality care. The new payment system should invest in the development of outcome measures to guide payment. 2. Establish a securely funded, independent agency to sponsor and evaluate research on the comparative effectiveness of drugs, devices, and other medical interventions. 3. Simplify and rationalize federal and state laws and regulations to facilitate organizational innovation, support care coordination, and streamline financial and administrative functions. 4. Develop a health information technology infrastructure with national standards of interoperability to promote data exchange. 5. Create a national health database with the participation of all payers, delivery systems, and others who own health care data. Agree on methods to make de-identified information from this database on clinical interventions, patient outcomes, and costs available to researchers. 6. Identify revenue sources, including a cap on the tax exclusion of employer-based health insurance, to subsidize health care coverage with the goal of insuring all Americans. 7. Create state or regional insurance exchanges to pool risk, so that Americans without access to employer-based or other group insurance could obtain a standard benefits package through these exchanges. Employers should also be allowed to participate in these exchanges for their employees' coverage. 8. Create a health coverage board with broad stakeholder representation to determine and periodically update the affordable standard benefit package available through state or regional insurance exchanges

Genome-wide examination of the transcriptional response to ecdysteroids 20-hydroxyecdysone and ponasterone A in Drosophila melanogaster

<p>Abstract</p> <p>Background</p> <p>The 20-hydroxyecdysone (20E) hierarchy of gene activation serves as an attractive model system for studying the mode of steroid hormone regulated gene expression and development. Many structural analogs of 20E exist in nature and among them the plant-derived ponasterone A (PoA) is the most potent. PoA has a higher affinity for the 20E nuclear receptor, composed of the ecysone receptor (EcR) and Ultraspiracle proteins, than 20E and a comparison of the genes regulated by these hormones has not been performed. Furthermore, in <it>Drosophila </it>different cell types elicit different morphological responses to 20E yet the cell type specificity of the 20E transcriptional response has not been examined on a genome-wide scale. We aim to characterize the transcriptional response to 20E and PoA in <it>Drosophila </it>Kc cells and to 20E in salivary glands and provide a robust comparison of genes involved in each response.</p> <p>Results</p> <p>Our genome-wide microarray analysis of Kc167 cells treated with 20E or PoA revealed that far more genes are regulated by PoA than by 20E (256 vs 148 respectively) and that there is very little overlap between the transcriptional responses to each hormone. Interestingly, genes induced by 20E relative to PoA are enriched in functions related to development. We also find that many genes regulated by 20E in Kc167 cells are not regulated by 20E in salivary glands of wandering 3^rdinstar larvae and we show that 20E-induced levels of <it>EcR </it>isoforms <it>EcR-RA, ER-RC</it>, and <it>EcR-RD/E </it>differ between Kc cells and salivary glands suggesting a possible cause for the observed differences in 20E-regulated gene transcription between the two cell types.</p> <p>Conclusions</p> <p>We report significant differences in the transcriptional responses of 20E and PoA, two steroid hormones that differ by only a single hydroxyl group. We also provide evidence that suggests that PoA induced death of non-adapted insects may be related to PoA regulating different set of genes when compared to 20E. In addition, we reveal large differences between Kc cells and salivary glands with regard to their genome-wide transcriptional response to 20E and show that the level of induction of certain EcR isoforms differ between Kc cells and salivary glands. We hypothesize that the differences in the transcriptional response may in part be due to differences in the EcR isoforms present in different cell types.</p

Molecular and atomic gas in dust lane early-type galaxies - I : Low star-formation efficiencies in minor merger remnants

In this work we present IRAM-30m telescope observations of a sample of bulge-dominated galaxies with large dust lanes, which have had a recent minor merger. We find these galaxies are very gas rich, with H2 masses between 4x10^8 and 2x10^10 Msun. We use these molecular gas masses, combined with atomic gas masses from an accompanying paper, to calculate gas-to-dust and gas-to-stellar mass ratios. The gas-to-dust ratios of our sample objects vary widely (between ~50 and 750), suggesting many objects have low gas-phase metallicities, and thus that the gas has been accreted through a recent merger with a lower mass companion. We calculate the implied minor companion masses and gas fractions, finding a median predicted stellar mass ratio of ~40:1. The minor companion likely had masses between ~10^7 - 10^10 Msun. The implied merger mass ratios are consistent with the expectation for low redshift gas-rich mergers from simulations. We then go on to present evidence that (no matter which star-formation rate indicator is used) our sample objects have very low star-formation efficiencies (star-formation rate per unit gas mass), lower even than the early-type galaxies from ATLAS3D which already show a suppression. This suggests that minor mergers can actually suppress star-formation activity. We discuss mechanisms that could cause such a suppression, include dynamical effects induced by the minor merger.Peer reviewe