Dual,Site‐Specific Modification of Antibodies by Using Solid‐Phase Immobilized Microbial Transglutaminase

Microbial transglutaminase (MTG) was stably solid‐phase immobilized on glass microbeads by using a second‐generation dendronized polymer. Immobilized MTG enabled the efficient generation of site‐specifically conjugated proteins, including antibody fragments, as well as whole antibodies through distinct glutamines and, unprecedentedly, also through lysines with various bifunctional substrates with defined stoichiometries. With this method, we generated dual, site‐specifically modified antibodies comprising a fluorescent probe and a metal chelator for radiolabeling—a strategy anticipated to design antibodies for imaging and simultaneous therapy. Furthermore, we provide evidence that immobilized MTG features higher siteselectivity than soluble MTG.     

Diving at diminished atmospheric pressure: air decompression tables for different altitudes

Fifty subjects performed 106 simulated dives at a final ambient pressure of 0.7 at (3000 m above sea level). One hundred and forty-three subjects performed 278 actual controlled dives at altitudes 900-1700 m above sea level. From the experience of these dives, air-decompression tables for altitudes 0-3200 m above sea level were calculated. Tables up to 2000 m above sea level were tested on humans under wet conditions.     

[18F]Flurpiridaz: Facile and Improved Precursor Synthesis for this Next-Generation Cardiac Positron Emission Tomography Imaging Agent

[¹⁸F]Flurpiridaz is a recently developed positron emission tomography tracer that is currently being investigated in phase III clinical trials to measure myocardial blood flow. The relatively long physical half-life of fluorine-18 alongside the high spatial resolution and outstanding myocardium-to-background ratio fuels its potential to be the next gold standard for the early detection of coronary artery disease. Notwithstanding the expected widespread use of [¹⁸F]flurpiridaz, the reported multistep synthesis of its precursor for radiofluorination involves a hazardous alkylation step using carcinogenic ethylene oxide, and a low overall chemical yield of 7 %. In this work, we have improved the overall yield more than fivefold and concurrently replaced the hazardous step. Specificity of binding of [¹⁸F]flurpiridaz to mitochondrial complex 1 was demonstrated by in vitro autoradiography on mouse heart tissue sections. These results thus pave the way for assessing myocardial blood flow and coronary flow reserve in mouse models of cardiovascular disease.     

A Click Approach to Structurally Diverse Conjugates Containing a Central Di‐1,2,3‐triazole Metal Chelate

Assemble & chelate : Click chemistry enables the efficient and selective synthesis of structurally diverse conjugates containing a central di‐1,2,3‐triazole chelator for complexation with [^99mTc(CO)₃]⁺. Use of appropriate building blocks allows the modulation of pharmacological relevant characteristics of the conjugate, or the introduction of secondary probes suitable for imaging modalities other than single photon emission computed tomography (SPECT).

     

Microbial Transglutaminase and c‐myc‐Tag: A Strong Couple for the Functionalization of Antibody‐Like Protein Scaffolds from Discovery Platforms

Antibody‐like proteins selected from discovery platforms are preferentially functionalized by site‐specific modification as this approach preserves the binding abilities and allows a side‐by‐side comparison of multiple conjugates. Here we present an enzymatic bioconjugation platform that targets the c‐myc‐tag peptide sequence (EQKLISEEDL) as a handle for the site‐specific modification of antibody‐like proteins. Microbial transglutaminase (MTGase) was exploited to form a stable isopeptide bond between the glutamine on the c‐myc‐tag and various primary‐amine‐functionalized substrates. We attached eight different functionalities to a c‐myc‐tagged antibody fragment and used these bioconjugates for downstream applications such as protein multimerization, immobilization on surfaces, fluorescence microscopy, fluorescence‐activated cell sorting, and in vivo nuclear imaging. The results demonstrate the versatility of our conjugation strategy for transforming a c‐myc‐tagged protein into any desired probe.     

Development of [(18)F]-PSS223 as a PET tracer for imaging of metabotropic glutamate receptor subtype 5 (mGluR5)

Involvement of metabotropic glutamate receptor subtype 5 (mGluR5) in physiological and pathophysiological processes in the brain has been demonstrated, and hence mGluR5 has emerged as an important drug target. [(11)C]-ABP688 is clinically the most successful mGluR5 positron emission tomography (PET) tracer to date and it allows visualization and quantification of mGluR5. Due to the short half-life of carbon-11, clinical use of [(11)C]-ABP688 is limited to facilities with an on-site cyclotron and a fluorine-18 (half-life 110 min) analogue would be more practical. Based on the [(11)C]-ABP688 structural motif, a novel derivative [(18)F]-PSS223 was prepared and evaluated as a PET tracer for imaging of mGluR5 in vitro and in vivo. Our results show favourable in vitro binding properties; however rapid defluorination of [(18)F]-PSS223 does not allow visualization of mGluR5 in the rat brain.     

Configurable front-end converters for multicurrent locomotives operated on 16 2/3 Hz AC and 3 kV DC systems

A configurable converter topology is presented as front-end converter for a locomotive that can be operated as well on a high voltage AC-system as on a medium voltage DC-system. The configurable converter is realized on the base of a multilevel chain, where all modules are fed through reversible and isolated DC-DC stages. The medium frequency transformers used in the isolation stages of the DC-DC converters allow a significant weight reduction of the locomotive, in comparison with the classical low-frequency voltage-transformers. A first solution using hard switching techniques is presented, using a three-times silicon-conversion. A second solution based on ZVT/ZCS techniques is then presented, with the advantage of having ideally only a two times silicon conduction. Prototype modules have been realized with association of diodes and IGBTs in order to provide reverse-blocking devices. First results of the two-stage conversion are also presented.     

A three-phase multilevel converter for high-power induction motors

A new converter topology for drives is presented in this paper: a three-phase multilevel converter with separately regulated DC power supplies. The DC voltages are provided by medium-frequency DC-DC converters. The applications for the converter are especially high-power traction systems, where the voltage applied to the induction motor is bigger than 1 kV. The motor current is of a very high quality, compared to a classical three-phase converter. This allows keeping the switching frequency low by using phase-shifted pulsewidth modulation (PWM) carriers. Different modulation methods have been developed and simulated. Experimental tests have been made on a 12 kW prototype     

An Electronc Circuit for Red/Infrared Oximeters

An inexpensive electronic circuit is described for operation of a red/infrared oximeter that does not use fiberoptics. The circuit provides low-evel amplitude modulation of the LED currents. Narrow-band detectors separate the backscattered signal into the red and infrared components. An AGC loop performs the division (Ei/Er).