Kopplung von Bilirubinoxidase an Kohlenstoffnanoröhrchen auf Elektroden

In der vorliegenden Arbeit wurde ein Elektrodensystem aus MWCNT-modifizierten Goldelektroden mit kovalent fixierter Bilirubinoxidase elektrochemisch untersucht. Unter aeroben Bedingungen konnte ein DET von der Elektrode zur BOD durch die bioelektrokatalytische Reduktion von Sauerstoff zu Wasser detektiert werden. Das Startpotential der Elektrodenreaktion unter aeroben Bedingungen wurde mit etwa 720±10mV gegen SHE ermittelt. Mit dem hier beschriebenen Elektrodensystem sind in luftgesättigtem Puffer (bei 25±5°C) Stromdichten im Bereich von etwa 500 μA/cm2 möglich. Durch Ultraschallbehandlung der Kohlenstoffnanoröhren vor der Präparation, Variation der Auftragungsmenge und der BOD-Konzentration während der Immobilisierung konnte eine Erhöhung des voltammetrischen Meÿsignals erzielt werden. Weiterhin war dieses Elektrodensystem bei physiologischem pH-Wert und relativ hoher Ionenstärke einsetzbar und erwies sich im Vergleich zu anderen Proteinelektroden mit bis zu 2 Monaten als sehr lagerstabil. Unter anaeroben Bedingungen wurde ein Kupferzentrum der Bilirubinoxidase einer direkten Redoxumwandlung unterworfen. Der Nachweis dieses Redoxprozesses gelang erstmalig mit kovalent fixierter Bilirubinoxidase. Bei pH 7 in 100mM CiP-Puffer mit 25mV/s wurde ein formales Potential von etwa 680±10mV gegen SHE bestimmt. Dieses Potential entspricht in etwa den in den letzten Jahren publizierten Standardpotentialen für das T1-Kupferzentrum von BOD.[1] Weitere Experimente mit Bilirubin und den Inhibitoren NaF und NaN3 unter aeroben und anaeroben Bedingungen deuteten ebenfalls daraufhin, daÿ beim DET mit BOD die Elektronen von der Elektrode auf das T1-Zentrum übertragen werden

Halbleiternanopartikel-modifizierte Elektrode zum Nachweis von Substraten von NADH-abhängigen Enzymreaktionen

Es wurde ein Elektrodensystem entwickelt, das aufbauend auf Halbleiternanopartikeln (so genannte Quantenpunkte) die sensitive Detektion des Enzymkofaktors NADH (nicotinamide adenine dinucleotide) erlaubt. Kolloidale halbleitende CdSe/ZnS-Nanokristalle sind durch ein Dithiol über Chemisorption an Gold gebunden. Das Stromsignal kann durch die Beleuchtung der Quantenpunkt modifizierten Oberfläche beeinflusst werden. Durch Photoanregung entstehen Elektron-Loch- Paare in den Nanopartikeln, die als anodischer oder kathodischer Photostrom detektiert werden können. Die Immobilisierung der Nanokristalle ist durch amperometrische Photostrom- und Quarzmikrowaage-Messungen (quartz crystal microbalance) verifiziert. Diese Studie zeigt, dass CdSe/ZnS-Quantenpunktmodifizierte Elektroden eine konzentrationsabhängige NADH-Detektion im Bereich von 20μM bis 2mM bei relativ niedrigem Potential (um 0V vs Ag/AgCl, 1 M KCl) ermöglichen. Somit können solche Elektroden in Kombination mit NADH-produzierenden Reaktionen für die lichtgesteuerte Analyse der entsprechenden Substrate des Biokatalysators genutzt werden. Es wird gezeigt, dass mit einem solchen Elektrodensystem und Photostrommessungen ein Glukosenachweis möglich ist.An electrode system based on semiconductive nanoparticles (so called quantum dots) was developed which allows the sensitive detection of the enzyme cofactor NADH (nicotinamide adenine dinucleotide). Colloidal semiconductive CdSe/ZnS nanocrystals are bound to gold via a dithiol compound by chemisorption. The current signal can be influenced by illumination of the quantum dot-modified electrode surface. Because of photoexcitation electron-holepairs are generated in the nanoparticles which can be detected as anodic or cathodic photocurrent. The immobilisation of the nanocrystals is verified by photocurrent and quarz crystal microbalance (QCM) measurements. This study shows that CdSe/ZnS-quatum dot-modified electrodes provide a concentration-dependent detection of NADH in the range of 20μM up to 2mM at relatively low overpotentials (around 0V vs Ag/ AgCl, 1 M KCl). Such electrodes can be used in combination with NADH-producing reactions for the lighttriggered analysis of the corresponding substrate of the biocatalyst. The detection of glucose with such an electrode system and photocurrent measurements is shown

Glutamic Acid Residues in HIV-1 p6 Regulate Virus Budding and Membrane Association of Gag

The HIV-1 Gag p6 protein regulates the final abscission step of nascent virions from the cell membrane by the action of its two late (l-) domains, which recruit Tsg101 and ALIX, components of the ESCRT system. Even though p6 consists of only 52 amino acids, it is encoded by one of the most polymorphic regions of the HIV-1 gag gene and undergoes various posttranslational modifications including sumoylation, ubiquitination, and phosphorylation. In addition, it mediates the incorporation of the HIV-1 accessory protein Vpr into budding virions. Despite its small size, p6 exhibits an unusually high charge density. In this study, we show that mutation of the conserved glutamic acids within p6 increases the membrane association of Pr55 Gag followed by enhanced polyubiquitination and MHC-I antigen presentation of Gag-derived epitopes, possibly due to prolonged exposure to membrane bound E3 ligases. The replication capacity of the total glutamic acid mutant E0A was almost completely impaired, which was accompanied by defective virus release that could not be rescued by ALIX overexpression. Altogether, our data indicate that the glutamic acids within p6 contribute to the late steps of viral replication and may contribute to the interaction of Gag with the plasma membrane

Methanotrophy under Versatile Conditions in the Water Column of the Ferruginous Meromictic Lake La Cruz (Spain)

Lakes represent a considerable natural source of methane to the atmosphere compared to their small global surface area. Methanotrophs in sediments and in the water column largely control methane fluxes from these systems, yet the diversity, electron accepting capacity, and nutrient requirements of these microorganisms have only been partially identified. Here, we investigated the role of electron acceptors alternative to oxygen and sulfate in microbial methane oxidation at the oxycline and in anoxic waters of the ferruginous meromictic Lake La Cruz, Spain. Active methane turnover in a zone extending well below the oxycline was evidenced by stable carbon isotope-based rate measurements. We observed a strong methane oxidation potential throughout the anoxic water column, which did not vary substantially from that at the oxic/anoxic interface. Both in the redox-transition and anoxic zones, only aerobic methane-oxidizing bacteria (MOB) were detected by fluorescence in situ hybridization and sequencing techniques, suggesting a close coupling of cryptic photosynthetic oxygen production and aerobic methane turnover. Additions of nitrate, nitrite and to a lesser degree iron and manganese oxides also stimulated bacterial methane consumption. We could not confirm a direct link between the reduction of these compounds and methane oxidation and we cannot exclude the contribution of unknown anaerobic methanotrophs. Nevertheless, our findings from Lake La Cruz support recent laboratory evidence that aerobic methanotrophs may be able to utilize alternative terminal electron acceptors under oxygen limitation

Role of Intraspecies Recombination in the Spread of Pathogenicity Islands within the Escherichia coli Species

Horizontal gene transfer is a key step in the evolution of bacterial pathogens. Besides phages and plasmids, pathogenicity islands (PAIs) are subjected to horizontal transfer. The transfer mechanisms of PAIs within a certain bacterial species or between different species are still not well understood. This study is focused on the High-Pathogenicity Island (HPI), which is a PAI widely spread among extraintestinal pathogenic Escherichia coli and serves as a model for horizontal transfer of PAIs in general. We applied a phylogenetic approach using multilocus sequence typing on HPI-positive and -negative natural E. coli isolates representative of the species diversity to infer the mechanism of horizontal HPI transfer within the E. coli species. In each strain, the partial nucleotide sequences of 6 HPI–encoded genes and 6 housekeeping genes of the genomic backbone, as well as DNA fragments immediately upstream and downstream of the HPI were compared. This revealed that the HPI is not solely vertically transmitted, but that recombination of large DNA fragments beyond the HPI plays a major role in the spread of the HPI within E. coli species. In support of the results of the phylogenetic analyses, we experimentally demonstrated that HPI can be transferred between different E. coli strains by F-plasmid mediated mobilization. Sequencing of the chromosomal DNA regions immediately upstream and downstream of the HPI in the recipient strain indicated that the HPI was transferred and integrated together with HPI–flanking DNA regions of the donor strain. The results of this study demonstrate for the first time that conjugative transfer and homologous DNA recombination play a major role in horizontal transfer of a pathogenicity island within the species E. coli

Global Health Education: a cross-sectional study among German medical students to identify needs, deficits and potential benefits (Part 1 of 2: Mobility patterns & educational needs and demands)

<p>Abstract</p> <p>Background</p> <p>In recent years, education and training in global health has been the subject of recurring debate in many countries. However, in Germany, there has been no analysis of the educational needs or demands of medical students, or the educational deficits or potential benefits involved in global health education. Our purpose is to analyse international health elective patterns of medical students enrolled at German universities and assess whether or how they prepare for their electives abroad. We examine the exposure of medical students enrolled at German universities to training courses in tropical medicine or global health and assess students' perceived needs and demands for education in global health.</p> <p>Methods</p> <p>Cross-sectional study among medical students in Germany including all 36 medical schools during the second half of the year 2007. All registered medical students were eligible to participate in the study. Recruitment occurred via electronic mailing-lists of students' unions. We developed a web-based, semi-structured questionnaire to capture students' international mobility patterns, preparation before electives, destination countries, exposure to and demand for global health learning opportunities.</p> <p>Results</p> <p>1126 online-replies were received and analysed from all registered medical students in Germany (N = 78.067). 33.0% of all respondents (370/1126) declared at least one international health elective and of these, 36.0% (133/370) completed their electives in developing countries. 36.0% (131/363) did not prepare specifically at all, 59.0% (214/363) prepared either by self-study or declared a participation in specific preparation programmes. 87.8% of 5^thand 6^thyear students had never participated in a global health course and 72.6% (209/288) had not completed a course in tropical medicine. 94.0% (861/916) endorsed the idea of introducing global health into medical education.</p> <p>Conclusion</p> <p>Students in our sample are highly mobile during their studies. International health electives are common, also in developing countries. Formal preparation beyond self-study is virtually non-existent amongst our sample and the participation rate in courses of tropical medicine or global health is appallingly low. We have identified unmet perceived needs and the demand for more learning opportunities in global health in our sample, urging for reforms to adjust curricula to a globalising world.</p

A USCLIVAR Project to Assess and Compare the Responses of Global Climate Models to Drought-Related SST Forcing Patterns: Overview and Results

The USCLI VAR working group on drought recently initiated a series of global climate model simulations forced with idealized SST anomaly patterns, designed to address a number of uncertainties regarding the impact of SST forcing and the role of land-atmosphere feedbacks on regional drought. Specific questions that the runs are designed to address include: What are the mechanisms that maintain drought across the seasonal cycle and from one year to the next? What is the role of the leading patterns of SST variability, and what are the physical mechanisms linking the remote SST forcing to regional drought, including the role of land-atmosphere coupling? The runs were carried out with five different atmospheric general circulation models (AGCM5), and one coupled atmosphere-ocean model in which the model was continuously nudged to the imposed SST forcing. This paper provides an overview of the experiments and some initial results focusing on the responses to the leading patterns of annual mean SST variability consisting of a Pacific El Nino/Southern Oscillation (ENSO)-like pattern, a pattern that resembles the Atlantic Multi-decadal Oscillation (AMO), and a global trend pattern. One of the key findings is that all the AGCMs produce broadly similar (though different in detail) precipitation responses to the Pacific forcing pattern, with a cold Pacific leading to reduced precipitation and a warm Pacific leading to enhanced precipitation over most of the United States. While the response to the Atlantic pattern is less robust, there is general agreement among the models that the largest precipitation response over the U.S. tends to occur when the two oceans have anomalies of opposite sign. That is, a cold Pacific and warm Atlantic tend to produce the largest precipitation reductions, whereas a warm Pacific and cold Atlantic tend to produce the greatest precipitation enhancements. Further analysis of the response over the U.S. to the Pacific forcing highlights a number of noteworthy and to some extent unexpected results. These include a seasonal dependence of the precipitation response that is characterized by signal-to-noise ratios that peak in spring, and surface temperature signal-to-noise ratios that are both lower and show less agreement among the models than those found for the precipitation response. Another interesting result concerns what appears to be a substantially different character in the surface temperature response over the U.S. to the Pacific forcing by the only model examined here that was developed for use in numerical weather prediction. The response to the positive SST trend forcing pattern is an overall surface warming over the world's land areas with substantial regional variations that are in part reproduced in runs forced with a globally uniform SST trend forcing. The precipitation response to the trend forcing is weak in all the models

Global Health Education: a cross-sectional study among German medical students to identify needs, deficits and potential benefits (Part 2 of 2: Knowledge gaps and potential benefits)

<p>Abstract</p> <p>Background</p> <p>In Germany, educational deficits or potential benefits involved in global health education have not been analysed till now.</p> <p>Objective</p> <p>We assess the importance medical students place on learning about social determinants of health (SDH) and assess their knowledge of global health topics in relation to (i) mobility patterns, their education in (ii) tropical medicine or (iii) global health.</p> <p>Methods</p> <p>Cross-sectional study among medical students from all 36 medical schools in Germany using a web-based, semi-structured questionnaire. Participants were recruited via mailing-lists of students' unions, all medical students registered in 2007 were eligible to participate in the study. We captured international mobility patterns, exposure to global health learning opportunities and attitudes to learning about SDH. Both an objective and subjective knowledge assessment were performed.</p> <p>Results</p> <p>1126 online-replies were received and analysed. International health electives in developing countries correlated significantly with a higher importance placed on all provided SDH (p ≤ 0.006). Participation in tropical medicine (p < 0.03) and global health courses (p < 0.02) were significantly associated with a higher rating of 'culture, language and religion' and the 'economic system'. Global health trainings correlated with significantly higher ratings of the 'educational system' (p = 0.007) and the 'health system structure' (p = 0.007), while the item 'politics' was marginally significant (p = 0.053).</p> <p>In the knowledge assessment students achieved an average score of 3.6 (SD 1.5; Mdn 4.0), 75% achieved a score of 4.0 or less (Q₂₅= 3.0; Q₇₅= 4.0) from a maximum achievable score of 8.0. A better performance was associated with international health electives (p = 0.032), participation in tropical medicine (p = 0.038) and global health (p = 0.258) courses.</p> <p>Conclusion</p> <p>The importance medical students in our sample placed on learning about SDH strongly interacts with students' mobility, and participation in tropical medicine and global health courses. The knowledge assessment revealed deficits and outlined needs to further analyse education gaps in global health. Developing concerted educational interventions aimed at fostering students' engagement with SDH could make full use of synergy effects inherent in student mobility, tropical medicine and global health education.</p

Enhancing precision in human neuroscience

Human neuroscience has always been pushing the boundary of what is measurable. During the last decade, concerns about statistical power and replicability – in science in general, but also specifically in human neuroscience – have fueled an extensive debate. One important insight from this discourse is the need for larger samples, which naturally increases statistical power. An alternative is to increase the precision of measurements, which is the focus of this review. This option is often overlooked, even though statistical power benefits from increasing precision as much as from increasing sample size. Nonetheless, precision has always been at the heart of good scientific practice in human neuroscience, with researchers relying on lab traditions or rules of thumb to ensure sufficient precision for their studies. In this review, we encourage a more systematic approach to precision. We start by introducing measurement precision and its importance for well-powered studies in human neuroscience. Then, determinants for precision in a range of neuroscientific methods (MRI, M/EEG, EDA, Eye-Tracking, and Endocrinology) are elaborated. We end by discussing how a more systematic evaluation of precision and the application of respective insights can lead to an increase in reproducibility in human neuroscience

New Strong-Field QED Effects at ELI: Nonperturbative Vacuum Pair Production

Since the work of Sauter, and Heisenberg, Euler and K\"ockel, it has been understood that vacuum polarization effects in quantum electrodynamics (QED) predict remarkable new phenomena such as light-light scattering and pair production from vacuum. However, these fundamental effects are difficult to probe experimentally because they are very weak, and they are difficult to analyze theoretically because they are highly nonlinear and/or nonperturbative. The Extreme Light Infrastructure (ELI) project offers the possibility of a new window into this largely unexplored world. I review these ideas, along with some new results, explaining why quantum field theorists are so interested in this rapidly developing field of laser science. I concentrate on the theoretical tools that have been developed to analyze nonperturbative vacuum pair production.Comment: 20 pages, 9 figures; Key Lecture at the ELI Workshop and School on "Fundamental Physics with Ultra-High Fields", 29 Sept - 2 Oct. 2008, Frauenworth Monastery, Germany; v2: refs updated, English translations of reviews of Nikishov and Ritu