Structural model of the CopA copper ATPase of Enterococcus hirae based on chemical cross-linking

The CopA copper ATPase of Enterococcus hirae belongs to the family of heavy metal pumping CPx-type ATPases and shares 43% sequence similarity with the human Menkes and Wilson copper ATPases. Due to a lack of suitable protein crystals, only partial three-dimensional structures have so far been obtained for this family of ion pumps. We present a structural model of CopA derived by combining topological information obtained by intramolecular cross-linking with molecular modeling. Purified CopA was cross-linked with different bivalent reagents, followed by tryptic digestion and identification of cross-linked peptides by mass spectrometry. The structural proximity of tryptic fragments provided information about the structural arrangement of the hydrophilic protein domains, which was integrated into a three-dimensional model of CopA. Comparative modeling of CopA was guided by the sequence similarity to the calcium ATPase of the sarcoplasmic reticulum, Serca1, for which detailed structures are available. In addition, known partial structures of CPx-ATPase homologous to CopA were used as modeling templates. A docking approach was used to predict the orientation of the heavy metal binding domain of CopA relative to the core structure, which was verified by distance constraints derived from cross-links. The overall structural model of CopA resembles the Serca1 structure, but reveals distinctive features of CPx-type ATPases. A prominent feature is the positioning of the heavy metal binding domain. It features an orientation of the Cu binding ligands which is appropriate for the interaction with Cu-loaded metallochaperones in solution. Moreover, a novel model of the architecture of the intramembranous Cu binding sites could be derive

Physical and Biogeochemical Studies in the Subtropical and Tropical Atlantic

Maria S. Merian Cruise Report MSM18/L2 Cruise No. 18, Leg 2 May 11 – June 19, 2011 Mindelo (Cape Verde Islands) – Mindelo (Cape Verde Islands

An advanced towed CTD chain for physical-biological high resolution in situ upper ocean measurements

Submesoscale eddies, fronts, and filaments are ubiquitous in the upper ocean and play an important role in biogeochemical and mixing processes as well as in the energy budget. To capture the high spatial variability of submesoscale processes, it is desirable to simultaneously resolve the vertical and horizontal gradients of hydrographic properties on scales of 10 m to 10 km. We present a revised towed CTD chain, for rapid quasi-synoptic in situ measurements of submesoscale oceanographic features, that is lighter, more robust and scientifically more useful than previous towed CTD chains. This new instrument provides a horizontal resolution of O(1 m) and can be towed at speeds of up to 5 ms-1 for measurements of the upper 100 m of the water column while providing a reasonable vertical resolution of O(1 m – 10 m). Individual CTD probes are equipped with temperature, conductivity, pressure and either rapid response dissolved oxygen or fluorescence sensors at multiple depths, enabling both hydrographic and biogeochemical studies at high resolution. A flexible probe hardware allows either real-time data collection or internal data logging for offline post-processing. Finally, we outline the necessary post-processing steps and provide data examples. With the presented data examples we show and conclude that the advanced towed CTD chain is a flexible and lightweight take on the towed CTD chain concept. It can easily be adapted to scientific needs and provides high quality very high resolution oceanographic data

Immunoadsorption for Treatment of Patients with Suspected Alzheimer Dementia and Agonistic Autoantibodies against Alpha1a-Adrenoceptor—Rationale and Design of the IMAD Pilot Study

Background: agonistic autoantibodies (agAABs) against G protein-coupled receptors (GPCR) have been linked to cardiovascular disease. In dementia patients, GPCR-agAABs against the α1- and ß2-adrenoceptors (α1AR- and ß2AR) were found at a prevalence of 50%. Elimination of agAABs by immunoadsorption (IA) was successfully applied in cardiovascular disease. The IMAD trial (Efficacy of immunoadsorption for treatment of persons with Alzheimer dementia and agonistic autoantibodies against alpha1A-adrenoceptor) investigates whether the removal of α1AR-AABs by a 5-day IA procedure has a positive effect (improvement or non-deterioration) on changes of hemodynamic, cognitive, vascular and metabolic parameters in patients with suspected Alzheimer’s clinical syndrome within a one-year follow-up period. Methods: the IMAD trial is designed as an exploratory monocentric interventional trial corresponding to a proof-of-concept phase-IIa study. If cognition capacity of eligible patients scores 19–26 in the Mini Mental State Examination (MMSE), patients are tested for the presence of agAABs by an enzyme-linked immunosorbent assay (ELISA)-based method, followed by a bioassay-based confirmation test, further screening and treatment with IA and intravenous immunoglobulin G (IgG) replacement. We aim to include 15 patients with IA/IgG and to complete follow-up data from at least 12 patients. The primary outcome parameter of the study is uncorrected mean cerebral perfusion measured in mL/min/100 gr of brain tissue determined by magnetic resonance imaging with arterial spin labeling after 12 months. Conclusion: IMAD is an important pilot study that will analyze whether the removal of α1AR-agAABs by immunoadsorption in α1AR-agAAB-positive patients with suspected Alzheimer’s clinical syndrome may slow the progression of dementia and/or may improve vascular functional parameters

Eddy Study to Understand Physical-Chemical-Biological Coupling and the Biological Carbon Pump as a Function of Eddy Type off West Africa, Cruise No. M160, 23.11.2019 - 20.12.2019, Mindelo (Cabo Verde) - Mindelo (Cabo Verde)

Cruise M160 is part of concerted MOSES/REEBUS Eddy Study featuring three major research expeditions (M156, M160, MSM104). It aims to develop both a qualitative and quantitative understanding of the role of physical-chemical-biological coupling in eddies for the biological pump. The study is part of the MOSES “Ocean Eddies” event chain, which follows three major hypotheses to be addressed by the MOSES/REEBUS field campaigns: (1) Mesoscale and sub-mesoscale eddies play an important role in transferring energy along the energy cascade from the large-scale circulation to dissipation at the molecular level. (2) Mesoscale and sub-mesoscale eddies are important drivers in determining onset, magnitude and characteristics of biological productivity in the ocean and contribute significantly to global primary production and particle export and transfer to the deep ocean. (3) Mesoscale and sub-mesoscale eddies are important for shaping extreme biogeochemical environments (e.g., pH, oxygen) in the oceans, thus acting as a source/sink function for greenhouse gases. In contrast to the other two legs, MOSES Eddy Study II during M160 did not include any benthic work but focused entirely on the pelagic dynamics within eddies. It accomplished a multi-disciplinary, multi-parameter and multi-platform study of two discrete cyclonic eddies in an unprecedented complexity. The pre-cruise search for discrete eddies suitable for detailed study during M160 had already started a few months prior to the cruise. Remote sensing data products (sea surface height, sea surface temperature, ocean color/chlorophyll a) were used in combination with eddy detection algorithms and numerical modelling to identify and track eddies in the entire eddy field off West Africa. In addition, 2 gliders and 1 waveglider had been set out from Mindelo/Cabo Verde for pre-cruise mapping of the potential working area north of the Cabo Verdean archipelago. At the start of M160, a few suitable eddies – mostly of cyclonic type – had been identified, some of which were outside the safe operation range of the motorglider plane. As technical problems delayed the flight operations, the first eddy (center at 14.5°N/25°W) for detailed study was chosen to the southwest of the island of Fogo. It was decided to carry out a first hydrographic survey there followed by the deployment of a suite of instruments (gliders, waveglider, floats, drifter short-term mooring). Such instrumented, we left this first eddy and transited – via a strong anticyclonic feature southwest of the island of Santiago – to the region northeast of the island of Sal, i.e. in the working range of the glider plane. During the transit, a full suite of underway measurements as well as CTD/RO section along 22°W (16°-18.5°N) were carried in search for sub-surface expressions of anticyclonic eddy features. In the northeast, we had identified the second strong cyclonic eddy (center at 18°N/22.5°W) which was chosen for detailed study starting with a complete hydrographic survey (ADCP, CTD/RO, other routine station work). After completion of the mesoscale work program, we identified a strong frontal region at the southwestern rim of the cyclonic eddy, which was chosen for the first sub-mesoscale study with aerial observation component. There, the first dye release experiment was carried out which consisted of the dye release itself followed by an intense multi-platforms study of the vertical and horizontal spreading of the initial dye streak. This work was METEOR-Berichte, Cruise M160, Mindelo – Mindelo, 23.11.2019 4 – 20.12.2019 supported and partly guided by aerial observation of the research motorglider Stemme, which was still somewhat compromised by technical issues and meteorological conditions (high cloud cover, Saharan dust event). Nevertheless, this first dye release experiment was successful and showed rapid movement of the dynamic meandering front. After completion of work on this second eddy and execution of a focused sampling program at the Cape Verde Ocean Observation, RV METEOR returned to the first eddy for continuation of the work started there in the beginning of the cruise. This was accompanied by a relocation of the airbase of Stemme from the international airport of Sal to the domestic airport of Fogo. The further execution of the eddy study at this first eddy, which again included a complete hydrographic survey followed by a mesoscale eddy study with dye release, was therefore possible with aerial observations providing important guidance for work on RV METEOR. Overall, M160 accomplished an extremely intense and complex work program with 212 instrument deployments during station work, 137 h of observation with towed instruments and a wide range of underway measurements throughout the cruise. Up to about 30 individually tracked platforms (Seadrones, glider, wavegliders, drifters, floats) were in the water at the same time providing unprecedented and orchestrated observation capabilities in an eddy. All planned work components were achieved and all working groups acquired the expected numbers of instrument deployments and sampling opportunities

A Connectionist Method To Solve Job Shop Problems

We propose a novel framework to solve job shop scheduling problems based on connectionist ideas of distributed information processing. In our approach each operation of a given job shop problem is considered to be a simple agent looking for a position in time such that all its time and resource constraints are satisfied. Each agent considers the current time position of its constraint neighbors to gradually change its own position to reach this goal. All agents together form a recurrent dynamical system which either self-organizes after some iterations to a feasible schedule or fails to do so depending on the constrainedness of the problem. By gradually increasing the constrainedness through decreasing the allowable overall processing time for a valid schedule, better and better solutions are found up to the point where no further improvements can be made. INTRODUCTION In classical job shop scheduling we are given a set of jobs, each of which consists of a chain of operations, and a s..

Strukturelle Charakterisierung der PDZ3-Domäne der PTP-BL und des Komplexes aus PDZ3-Domäne und dem carboxy-terminalen Dodecapeptid der PRK2

PDZ-Domänen sind weit verbreitete globuläre Protein-Protein-Interaktionsdomänen, die bevorzugt kurze, lineare, meist carboxy-terminale Motive ihrer Ligandproteine selektiv erkennen und binden können. Sie weisen eine konservierte $\alpha$ / $\beta$-Faltung, bestehend aus fünf bis sechs $\beta$-Faltblattsträngen und zwei $\alpha$-Helices, auf, und binden ihre Liganden in einer Furche zwischen $\beta$B-Strang und $\alpha$B-Helix. Essentiell für die Interaktion sind die vier carboxy-terminalen Aminosäuren des Bindungsmotivs des Liganden. In der vorliegenden Arbeit konnte die Struktur der PDZ3-Domäne der PTP-BL sowohl in der Peptid-freien Apoform als auch in der Komplexform gelöst werden. Als Ligand wurde ein Dodecapeptid verwendet, welches den Carboxy-Terminus des natürlichen Liganden der PDZ3-Domäne der PTP-BL, der PRK2, nachahmt