Prevalence and clinical impact of endoscopic pseudomembranes in patients with inflammatory bowel disease and Clostridium difficile infection

Background and aim: Limited data suggests that pseudomembranes are uncommon in patients with inflammatory bowel disease (IBD) and C. difficile associated disease (CDAD), but the reason for this is unknown. We aimed to evaluate the rate of pseudomembranes in this population, identify predictive factors for pseudomembranes' presence and assess its clinical impact. Methods: This was a sub-study of a retrospective European Crohn's & Colitis Organization (ECCO) multi-center study on the outcome of hospitalized IBD patients with C. difficile. The present study included only patients who underwent lower endoscopy during hospitalization, and compared demographic and clinical parameters in the group of patients with discernable pseudomembranes versus those without. Results: Out of 155 patients in the original cohort, 93 patients underwent lower endoscopy and constituted the study population. Endoscopic pseudomembranes were found in 12 (13%) of these patients. Patients with pseudomembranes presented more commonly with fever (p=0.02) compared to patients without pseudomembranes. No difference between the two groups was found with respect to the use of immunosuppressant drugs, background demographics or disease characteristics. Neither was there a difference between the group with or without pseudomembranes in the frequency of severe adverse clinical outcome or in the duration of hospitalization. On multi-variate analysis the presence of fever remained independently associated with the finding of pseudomembranes (OR 6, 95% CI 1.2–32, p=0.03). Conclusions: This study documents that hospitalized IBD patients with CDAD have low rate of endoscopic pseudomembranes, which is not accounted for by the use of immunosuppressant drugs. IBD patients with CDAD and discernable pseudomembranes more commonly present with fever, but their clinical outcome is similar to patients without pseudomembranes. © 2009 Published by Elsevier B.V. on behalf of European Crohn's and Colitis Organisation

Ortsaufgelöster Nachweis von Elementspuren mittels Laserablation und Resonanzionisations-Massenspektrometrie und Untersuchungen zur Laserablations-Dynamik

Im Rahmen dieser Arbeit wurde eine neue Methode für einen empfindlichen und isotopenselektiven Elementnachweis entwickelt. Unter Einsatz von Laserablation geschieht der Probenaufschluß direkt und mit einer Ortsauflösung von unter 30 m. Hierzu wurde ein hochauflösendes MALDI-TOF-Massenspektrometer, welches üblicherweise für biochemische Fragestellungen eingesetzt wird, mit einem spektroskopischen Aufbau zur resonanten Ionisation von Elementgehalten modifiziert. Die Methode ist somit insbesondere für die Untersuchung von Elementspuren in Festkörperproben mit mikroskopischer Struktur konzipiert. Methodische Entwicklungsarbeiten wurden anhand des Elements Gadolinium durchgeführt. Durch die Verwendung gepulster Farbstofflaser stehen ausreichend hohe Laserfelder zur Verfügung, um unabhängig von Hyperfeinstruktur und Isotopieverschiebung Übergänge aller Isotope im Rahmen des Resonanzionisations-Verfahrens zu sättigen. Darauf konnte eine Isotopenverhältnisanalyse mit einer Genauigkeit im Prozentbereich verwirklicht werden. Verschiedene Anregungsleitern wurden untersucht, und mit elementspezifischen Resonanzüberhöhungen bis zu zwei Größenordnungen über dem nicht-resonant gebildeten Untergrund konnte eine Nachweiseffizienz von über 10-4 (entsprechend sub-fg/g-Niveau) erzielt werden. Dazu wurden Simulationsrechnungen zum atomaren Sättigungsverhalten in starken resonanten Laserfeldern durchgeführt. Erste Anwendungen des Laserablationsverfahrens waren Proben kosmologischer Herkunft. Der physikalische Prozeß der Laserablation bei Metallen wurde unter Hochvakuum-Bedingung systematisch in Abhängigkeit der Laserfluenz untersucht. In der ablatierten Plasmaphase erwies sich der Neutralanteil als besonders geeignet für geschwindigkeitsselektive Laserionisations-Messungen. Eine bimodale Struktur wurde beobachtet, bestehend aus einer thermischen und einer schockwellen-induzierten Komponente. Der ionische Anteil der ablatierten Dampfphase konnte über variable elektrische Feldpulse untersucht werden. Laserablation unter Atmosphärenbedingung wurde an einem beschichteten Messingtarget untersucht. Dabei wurde die Entstehung von permanenten Oberflächenstrukturen beobachtet, welche sich durch Nichtgleichgewichts-Prozesse in der Dampfphase erklären lassen.Within the present work, a novel method for a sensitive and isotope selective detection of elemental traces has been developed. The sample decomposition is thereby realized by laser ablation, with a spatial resolution below 30 m. Therefore, a high resolution MALDI-TOF mass spectrometer, which is normally used for biochemical applications, has been combined with a spectroscopic setup for the resonant ionization of elements. This method is especially conceived for the investigations of elemental traces in solid samples with microscopic structure, where a conventional chemical treatment fails because of possible contaminations. By the use of pulsed dye lasers, sufficient laser field strengths are available for transition saturations of all isotopes of the test element gadolinium within the resonance ionization procedure, and an isotopic ratio analysis at a percent level of accuracy is realized. Different spectroscopic excitation schemes have been investigated, and an element specific resonance enhancement up to two orders of magnitude from non-resonant background has been obtained. Thereby, the detection efficiency exceeds a value of 10-4 (corresponding to sub-fg/g-level). Additional simulation work has been carried out, mainly to describe the atomic saturation behavior under intense resonant laser fields. Subsequently, the laser ablation method has been applied to samples of cosmological relevance. In an additional section, physical processes during laser ablation of metallic substrates have been investigated. After analyzing the composition of the ablated phase in dependency of the applied ablation laser fluence, the neutral fraction has been used for a velocity selective laser ionization measurement. A bimodal structure has emerged, indicating the presence of a thermal and shockwave-driven component. The ionic part of the ablation plume has been investigated by means of controled electric field manipulation of ion energy classes. Additional investigations have been performed on the base of a coated brass target under atmospheric condition, where nonequilibrium phenomena during the ablation process could be directly visualized from the remaining surface morphology

Bimodal velocity distribution of atoms released from nanosecondultraviolet laser ablation

We have investigated the velocity distributions of atoms released from a metallic gadolinium surface by UV laser ablation. The fluences of the nanosecond laser pulses were chosen for a pure release of neutrals and at a higher fluence level for the release of both neutrals and ions. In both cases a thermal Maxwell-Boltzmann slope has been observed for the low velocities, whereas for high velocities strong deviations from a thermal distribution have been seen. The observed velocity distribution has been explained by a bimodal structure including a thermal phase and a shockwave driven “blow-off” phase

Spatially resolved ultra-trace analysis of elements combining resonanceionization with a MALDI-TOF spectrometer

A combined setup for spatially resolved mass analysis of trace amounts of elements and macromolecules is presented. Using a MALDI-TOF mass spectrometer, a laser spectroscopic setup for resonant ionization of neutral atoms has been implemented. This allows for an efficient and selective detection of trace elements by means of resonance ionization mass spectrometry (RIMS). The instrumental scheme is described, and methodological developments are presented. In a first application pure, laser desorption/ionization with TOF-MS was used to measure mass distributions of cosmic nanodiamonds. For further applications regarding the spatially resolved ultra-trace analysis of elements in solid samples, an implanted target was used to characterize both laser desorption/ionization and laser desorption/resonance ionization for the detection of trace elements within. A perspective of the setup is given and future investigations are outlined

A laser desorption/resonance enhanced photoionisation TOF-system forthe spatially resolved trace analysis of elements

A novel method for direct and spatially resolved elemental trace analysis with high sensitivity and elemental selectivity is presented. The concept is based on the combination of a commercial MALDI-TOF mass spectrometer with a pulsed laser system for resonant postionisation of neutrals. While the MALDI method is usually applied for investigations of large organic compounds and biomolecules, the technique discussed here concerns the low mass range around 1 less than or equal to A less than or equal to 300. The analytical performances of the setup with respect to mass analysis, spatial resolution and overall detection efficiency are discussed. (C) 2004 Elsevier B.V. All rights reserved

Water Influences on the Copper Active Site in Hemocyanin

Active metal sites play a key role in the biochemistry of oxygen transport by hemocyanins. Observing the changes in the local electronic structure of the copper sites upon oxygenation is thus essential for understanding their biological functionality. Here, direct access to the electronic structure of the active copper sites in hemocyanin is achieved via L-edge X-ray absorption spectroscopy under physiological conditions. We compare the deoxygenated and the oxygenated states of native hemocyanin and find evidence that the oxygenation does not simply switch the copper valence state between Cu I and Cu II, as assumed classically. In the deoxygenated state, water molecules can enter the active site and keep the copper atoms partially oxidized. The role of water in this process has never been revealed before for lack of L-edge spectroscopy on copper in solution. Besides providing a more detailed electronic picture for the oxygenation process, this study opens a new chapter in investigating the function of proteins under in-vivo-like conditions

H1N1 vaccines in a large observational cohort of patients with inflammatory bowel disease treated with immunomodulators and biological therapy.

BACKGROUND: Safety data are lacking on influenza vaccination in general and on A (H1N1)v vaccination in particular in patients with inflammatory bowel disease (IBD) receiving immmunomodulators and/or biological therapy. AIMS AND METHODS: The authors conducted a multicentre observational cohort study to evaluate symptoms associated with influenza H1N1 adjuvanted (Pandemrix, Focetria, FluvalP) and non-adjuvanted (Celvapan) vaccines and to assess the risk of flare of IBD after vaccination. Patients with stable IBD treated with immunomodulators and/or biological therapy were recruited from November 2009 until March 2010 in 12 European countries. Harvey-Bradshaw Index and Partial Mayo Score were used to assess disease activity before and 4 weeks after vaccination in Crohn's disease (CD) and ulcerative colitis (UC). Vaccination-related events up to 7 days after vaccination were recorded. RESULTS: Of 575 patients enrolled (407 CD, 159 UC and nine indeterminate colitis; 53.9% female; mean age 40.3 years, SD 13.9), local and systemic symptoms were reported by 34.6% and 15.5% of patients, respectively. The most common local and systemic reactions were pain in 32.8% and fatigue in 6.1% of subjects. Local symptoms were more common with adjuvanted (39.3%) than non-adjuvanted (3.9%) vaccines (p < 0.0001), whereas rates of systemic symptoms were similar with both types (15.0% vs 18.4%, p = 0.44). Among the adjuvanted group, Pandemrix more often induced local reactions than FluvalP and Focetria (51.2% vs 27.6% and 15.4%, p < 0.0001). Solicited adverse events were not associated with any patient characteristics, specific immunomodulatory treatment, or biological therapy. Four weeks after vaccination, absence of flare was observed in 377 patients with CD (96.7%) and 151 with UC (95.6%). CONCLUSION: Influenza A (H1N1)v vaccines are well tolerated in patients with IBD. Non-adjuvanted vaccines are associated with fewer local reactions. The risk of IBD flare is probably not increased after H1N1 vaccination