A unique protein profile of peripheral neutrophils from COPD patients does not reflect cytokine-induced protein profiles of neutrophils in vitro

Contains fulltext : 96603.pdf (publisher's version ) (Open Access)ABSTRACT: BACKGROUND: Inflammation, both local and systemic, is a hallmark of chronic obstructive pulmonary disease (COPD). Inflammatory mediators such as TNFalpha and GM-CSF are secreted by lung epithelium, alveolar macrophages and other inflammatory cells and are thought to be important contributors in the pathogenesis of COPD. Indeed, neutrophils are activated by these cytokines and these cells are one of the major inflammatory cell types recruited to the pulmonary compartment of COPD patients. Furthermore, these inflammatory mediators are found in the peripheral blood of COPD patients and, therefore, we hypothesized that TNFalpha/GM-CSF-induced protein profiles can be found in peripheral neutrophils of COPD patients. METHODS: Using fluorescence 2-dimensional difference gel electrophoresis we investigated differentially regulated proteins in peripheral neutrophils from COPD patients and healthy age-matched control subjects. Furthermore, protein profiles from COPD patients were compared with those of neutrophils of healthy age-matched controls that were stimulated with TNFalpha and/or GM-CSF in vitro. Protein gels were compared using DeCyder 7.0 software. RESULTS: We identified 7 significantly regulated protein spots between peripheral neutrophils from COPD patients and age-matched healthy control subjects. Stimulation of peripheral neutrophils with TNFalpha, GM-CSF or TNFalpha + GM-CSF in vitro resulted in 13, 20 and 22 regulated protein spots, respectively. However, these cytokine-induced protein differences did not correspond with the protein differences found in neutrophils from COPD patients. CONCLUSION: These results show that neutrophils from COPD patients have a unique protein profile compared to neutrophils from healthy age-matched controls. Furthermore, the neutrophil profiles of COPD patients do not reflect putative dominant signals induced by TNFalpha, GM-CSF or their combination. Our results indicate that systemic neutrophil responses in COPD patients are caused by a unique but subtle interplay between multiple inflammatory signals

In-situ Infrared Characterization During Atomic Layer Deposition of Lanthanum Oxide

Mechanisms of atomic layer deposition (ALD) growth of lanthanum oxide on H-terminated Si(111) using lanthanum tris(N,N′-diisopropylacetamidinate) (La(iPr-MeAMD)3) are investigated using infrared (IR) absorption spectroscopy. The reactivity of this amidinate precursor is high, with almost all surface Si−H bonds consumed after 5 ALD cycles at 300 °C. Gas phase IR spectra show that, although most of the precursor (La(iPr-MeAMD)3) remains intact, a strong feature at 1665 cm−1, characteristic of a hydrogenated and dissociated free ligand with localized electrons in the N−CN bonds, is present. Such partial precursor dissociation in the gas phase is due to hydrolysis by traces of water vapor remaining in the reactor, even after purging. As a result, some Si−O−La bonds are formed upon reaction with the surface during the first La(iPr-MeAMD)3 pulse, prior to any water pulse. During film growth, acetate/carbonate and hydroxyl impurities are incorporated into the film. Annealing to 500 °C in dry N2 removes these impurities but fosters the growth of interfacial SiO2. Deposition at 300 °C leads to decomposition of adsorbed ligands, as evidenced by the formation of cyanamide or carbodiimide vibrational bands (or both) at 1990 and 2110 cm−1, respectively. Despite this decomposition, ideal self-limited ALD growth is maintained because the decomposed ligands are removed by the subsequent water pulse. Growth of pure lanthanum oxide films is often characterized by nonuniform film thickness if purging is not complete because of reversible absorption of water by the La2O3 film. Uniform ALD growth can be maintained without a rigorous dry purge by introducing alternating trimethylaluminum (TMA)/D2O ALD cycles between La/D2O cycles.Chemistry and Chemical Biolog

Late Neogene chronology: New perspectives in high-resolution stratigraphy

We present an integrated geochronology for late Neogene time (Pliocene, Pleistocene, and Holocene Epochs) based on an analysis of data from stable isotopes, magnetostratigraphy, radiochronology, and calcareous plankton biostratigraphy. Discrepancies between recently formulated astronomical chronologies and magnetochronologies for the past 6 m.y. have been resolved on the basis of new, high-precision Ar/Ar ages in the younger part of this interval, the so-called Brunhes, Matuyama, and Gauss Epochs (= Chrons C1n-C2An; 0-3.58 Ma), and revised analysis of sea floor anomalies in the Pacific Ocean in the older part, the so-called Gilbert Epoch (= Chron C2Ar-C3r; 3.58-5.89 Ma). The magneto- and astrochronologies are now concordant back to the Chron C3r/C3An boundary at 5.89 Ma. The Neogene (Miocene, Pliocene, Pleistocene, and Holocene) and Paleogene are treated here as period/system subdivisions of the Cenozoic Era/Erathem, replacements for the antiquated terms Tertiary and Quaternary. The boundary between the Miocene and Pliocene Series (Messinian/Zanclean Stages), whose global stratotype section and point (GSSP) is currently proposed to be in Sicily, is located within the reversed interval just below the Thvera (C3n.4n) Magnetic Polarity Subchronozone with an estimated age of 5.32 Ma. The Pliocene/Pleistocene boundary, whose GSSP is located at Vrica (Calabria, Italy), is located near the top of the Olduvai (C2n) Magnetic Polarity Subchronozone with an estimated age of 1.81 Ma. The 13 calcareous nannoplankton and 48 planktonic foraminiferal datum events for the Pliocene, and 12 calcareous nannoplankton and 10 planktonic foraminiferal datum events for the Pleistocene, are calibrated to the newly revised late Neogene astronomical/geomagnetic polarity time scale

Post-Eocene coupled oroclines in the Talesh (NW Iran): paleomagnetic constraints

The Talesh Mountains (NW Iran) witnessed a long deformation history from the Triassic Cimmerian orogeny to the ongoing Arabia-Eurasia collision. This protracted multi-stage deformation has generated a remarkably curved orogen with a puzzling kinematic and deformational history. In this study, we investigate the origin of the Talesh curvature through paleomagnetic analyses on rocks of Paleozoic, Mesozoic and Cenozoic age. Our results indicate that at least two major, large-scale, vertical-axis-rotations took place since the Late Cretaceous: 1) a pre-Eocene 73° ± 17° clockwise rotation and 2) post-Eocene differential rotations that formed the Z-shaped mountain belt within a crustal-scale shear zone. The latter involved an increasing amount of clockwise (CW) rotation from south (16°) to north (48°). The orocline formation likely started during the Oligocene where an approximately east-west oriented mountain belt was buckled by the Arabia-Eurasia collision, with Arabia acting as a rigid indenter and the South Caspian basin as a rigid backstop. We hypothesise that the NE-SW oriented Aras and Lahijan fault zones, inherited from transform faults related to the Mesozoic opening of the Caspian-Black Sea back-arc, accommodated the coupled orocline formation

Examining the distribution and impact of single nucleotide polymorphisms in the capsular locus of Streptococcus pneumoniae serotype 19A

Streptococcus pneumoniae serotype 19A prevalence has increased after implementation of PCV7 and PCV10 vaccines. In this study, we have provided, with high accuracy, the genetic diversity of the 19A serotype in a cohort of Dutch invasive pneumococcal disease patients and asymptomatic carriers obtained in the period 2004-2016. Whole genomes of the 338 pneumococcal isolates in this cohort were sequenced and their capsule (cps) loci compared to examine the diversity and determine the impact on the production of CPS sugar precursors and CPS shedding. We discovered 79 types with a unique CPS locus sequence. Most variation was observed in the rmlB and rmlD genes of the TDP-Rha synthesis pathway, and in the wzg gene, of unknown function. Interestingly, gene variation in the cps locus was conserved in multiple alleles. Using RmlB and RmlD protein models, we predict that enzymatic function is not affected by the single nucleotide polymorphisms as identified. To determine if RmlB and RmlD function was affected, we analyzed nucleotide sugar levels using UHPLC-MS. CPS precursors differed between 19A cps locus subtypes, including TDP-Rha, but no clear correlation was observed. Also, a significant difference in multiple nucleotide sugar levels was observed between phylogenetically branched groups. Because of indications of a role for Wzg in capsule shedding, we analyzed if this was affected. No clear indication of a direct role in shedding was found. We thus describe genotypic variety in rmlB, rmlD and wzg in serotype 19A the Netherlands, for which we have not discovered an associated phenotype

Probing Tectonic Topography in the Aftermath of Continental Convergence in Central Europe

Continental topography is at the interface of processes taking place at depth in the Earth,at its surface,and above it.Topography influences society, not only in terms of slow processes of landscape change and earthquakes,but also in terms of how it affects climate.The Pannonian Basin–Carpathian Orogen System in Central and Eastern Europe represents a key natural laboratory for the development of a new generation of models for ongoing orogeny and its effect on continental topography development (Figure 1).This system comprises some of the best documented sedimentary basins in the world,located within the Alpine orogenic belt, at the transition between the western European lithosphere and the East European Craton. It includes one of the most active seismic zones in Europe,with intermediate depth (50–220 km) mantle earthquakes of significant magnitude occurring in a geographically restricted area in the Vrancea zone of southeastern Romania

Cigarette smoke-exposed neutrophils die unconventionally but are rapidly phagocytosed by macrophages

Pulmonary accumulation of neutrophils is typical for active smokers who are also predisposed to multiple inflammatory and infectious lung diseases. We show that human neutrophil exposure to cigarette smoke extract (CSE) leads to an atypical cell death sharing features of apoptosis, autophagy and necrosis. Accumulation of tar-like substances in autophagosomes is also apparent. Before detection of established cell death markers, CSE-treated neutrophils are effectively recognized and non-phlogistically phagocytosed by monocyte-derived macrophages. Blockade of LOX-1 and scavenger receptor A, but not MARCO or CD36, as well as pre-incubation with oxLDL, inhibited phagocytosis, suggesting that oxLDL-like structures are major phagocytosis signals. Specific lipid (β-carotene and quercetin), but not aqueous, antioxidants increased the pro-phagocytic effects of CSE. In contrast to non-phlogistic phagocytosis, degranulation of secondary granules, as monitored by lactoferrin release, was apparent on CSE exposure, which is likely to promote pulmonary inflammation and tissue degradation. Furthermore, CSE-exposed neutrophils exhibited a compromised ability to ingest the respiratory pathogen, Staphylococcus aureus, which likely contributes to bacterial persistence in the lungs of smokers and is likely to promote further pulmonary recruitment of neutrophils. These data provide mechanistic insight into the lack of accumulation of apoptotic neutrophil populations in the lungs of smokers and their increased susceptibility to degradative pulmonary diseases and bacterial infections