The Deubiquitinase OTUB1 Is a Key Regulator of Energy Metabolism

Dysregulated energy metabolism is a major contributor to a multitude of pathologies, including obesity and diabetes. Understanding the regulation of metabolic homeostasis is of utmost importance for the identification of therapeutic targets for the treatment of metabolically driven diseases. We previously identified the deubiquitinase OTUB1 as substrate for the cellular oxygen sensor factor-inhibiting HIF (FIH) with regulatory effects on cellular energy metabolism, but the physiological relevance of OTUB1 is unclear. Here, we report that the induced global deletion of OTUB1 in adult mice (Otub1 iKO) elevated energy expenditure, reduced age-dependent body weight gain, facilitated blood glucose clearance and lowered basal plasma insulin levels. The respiratory exchange ratio was maintained, indicating an unaltered nutrient oxidation. In addition, Otub1 deletion in cells enhanced AKT activity, leading to a larger cell size, higher ATP levels and reduced AMPK phosphorylation. AKT is an integral part of insulin-mediated signaling and Otub1 iKO mice presented with increased AKT phosphorylation following acute insulin administration combined with insulin hypersensitivity. We conclude that OTUB1 is an important regulator of metabolic homeostasis.     

Short communication: Evaluation of an automated in-house hematology analyzer for bovine blood

The objective of this study was to evaluate the suitability of the V-Sight hematology analyzer (A. Menarini Pharma GmbH, Vienna, Austria) for bovine blood by a comparison with a reference device (Advia 2120i, Siemens AG, Erlangen, Germany). In total, 97 blood samples were obtained from 75 dairy cows. Analyzed parameters included counts of white blood cells (WBC), lymphocytes, monocytes, granulocytes, red blood cells (RBC), and platelets (PLT), as well as hemoglobin concentration (HGB), hematocrit (HCT), mean corpuscular volume, mean corpuscular hemoglobin (MCH), MCH concentration (MCHC), mean platelet volume (MPV), and plateletcrit (PCT). Based on Passing-Bablok regression, the V-Sight provided accurate and precise results for MCH and MCHC only. The PCT results were comparable to the reference method, but precision was inconclusive. Significant proportional differences were detected for monocytes, granulocytes, HCT, and PLT. For all other analytes, significant proportional and systemic differences were observed. The WBC and lymphocyte results from the V-Sight were characterized by poor accuracy, poor precision, and a high number of false positive outliers. Bland-Altman analysis indicated negative biases for all WBC parameters, the erythrocyte indices, and PLT. Positive biases were observed for RBC, HGB, HCT, MPV, and PCT. Correlation coefficients of >0.9 between the V-Sight and the reference method were found only for RBC, HGB, HCT, and MPV. Intraassay precision of the V-Sight analyzer was acceptable (coefficient of variation <5%) for granulocytes, the erythrocyte indices, and MPV. It was unacceptable (coefficient of variation ≥5%) for WBC, lymphocytes, monocytes, as well as RBC, and inconclusive for HGB, HCT, PLT, and PCT. Sensitivity was high for all RBC counts and indices as well as PLT, but low for monocytes, granulocytes, and MPV. Specificity was high for monocytes and granulocytes, but low for RBC, HCT, MCH, and MCHC. With accurate and precise results for only 2 out of 13 parameters, the V-Sight cannot be recommended for analysis of bovine blood.     

Wind energy potential in southern Sweden-Example of planning methodology

This study presents a planning model for Swedish activities in the field of wind power. Models and results of calculations of the land-based wind energy potential with the Wind Atlas Analysis and Application Programme, WA^sP, are described and analysed in a geographical information systems (GIS) called the ArcView^® GIS system. One county in southern Sweden was chosen as a case study to present the methods used. The results from that case study indicate a great wind energy potential but there are nevertheless many factors limiting that potential. The study calls for a further development of planning tools in the field. Important areas for the future are the development of knowledge in market issues, wind power technology, environmental issues, and public opinion on constructing wind turbines.     

Optimal sensor placement for joint parameter and state estimation problems in large-scale dynamical systems with applications to thermo-mechanics

We consider large-scale dynamical systems in which both the initial state and some parameters are unknown. These unknown quantities must be estimated from partial state observations over a time window. A data assimilation framework is applied for this purpose. Specifically, we focus on large-scale linear systems with multiplicative parameter-state coupling as they arise in the discretization of parametric linear time-dependent partial differential equations. Another feature of our work is the presence of a quantity of interest different from the unknown parameters, which is to be estimated based on the available data. In this setting, we employ a simplicial decomposition algorithm for an optimal sensor placement and set forth formulae for the efficient evaluation of all required quantities. As a guiding example, we consider a thermo-mechanical PDE system with the temperature constituting the system state and the induced displacement at a certain reference point as the quantity of interest.     

Nitroreductase-Mediated Release of Inhibitors of Lysine-Specific Demethylase 1 (LSD1) from Prodrugs in Transfected Acute Myeloid Leukaemia Cells

Lysine-specific demethylase 1 (LSD1) has evolved as a promising therapeutic target for cancer treatment, especially in acute myeloid leukaemia (AML). To approach the challenge of site-specific LSD1 inhibition, we developed an enzyme-prodrug system with the bacterial nitroreductase NfsB (NTR) that was expressed in the virally transfected AML cell line THP1-NTR⁺. The cellular activity of the NTR was proven with a new luminescent NTR probe. We synthesised a diverse set of nitroaromatic prodrugs that by design do not affect LSD1 and are reduced by the NTR to release an active LSD1 inhibitor. The emerging side products were differentially analysed using negative controls, thereby revealing cytotoxic effects. The 2-nitroimidazolyl prodrug of a potent LSD1 inhibitor emerged as one of the best prodrug candidates with a pronounced selectivity window between wild-type and transfected THP1 cells. Our prodrugs are selectively activated and release the LSD1 inhibitor locally, proving their suitability for future targeting approaches.     

Regression Techniques for Oceanographic Parameter Retrieval Using Space-Borne Microwave Radiometry

Variations of conventional multiple regression techniques are applied to the problem of remote sensing of oceanographic parameters from space. The techniques are specifically adapted to the scanning multichannel microwave radiometer (SMMR) launched on the Seasat and Nimbus 7 satellites to determine ocean surface temperature, wind speed, and atmospheric water content. The retrievals are studied primarily from a theoretical viewpoint, to illustrate the retrieval error structure, the relative importances of different radiometer channels, and the tradeoffs between spatial resolution and retrieval accuracy. Comparisons between regressions using simulated and actual SMMR data are discussed; they show similar behavior.     

Pressure cell for investigations of solid-liquid interfaces by neutron reflectivity

We describe an apparatus for measuring scattering length density and structure of molecular layers at planar solid-liquid interfaces under high hydrostatic pressure conditions. The device is designed for in situ characterizations utilizing neutron reflectometry in the pressure range 0.1-100 MPa at temperatures between 5 and 60?°C. The pressure cell is constructed such that stratified molecular layers on crystalline substrates of silicon, quartz, or sapphire with a surface area of 28 cm(2) can be investigated against noncorrosive liquid phases. The large substrate surface area enables reflectivity to be measured down to 10(-5) (without background correction) and thus facilitates determination of the scattering length density profile across the interface as a function of applied load. Our current interest is on the stability of oligolamellar lipid coatings on silicon surfaces against aqueous phases as a function of applied hydrostatic pressure and temperature but the device can also be employed to probe the structure of any other solid-liquid interface.