A New Colorimetric Assay for Methionyl Aminopeptidases: Examination of the Binding of a New Class of Pseudopeptide Analog Inhibitors

A direct and convenient spectrophotometric assay has been developed for methionine aminopeptidases (MetAPs). The method employs the hydrolysis of a substrate that is a methionyl analogue of p-nitroaniline (l-Met-p-NA), which releases the chromogenic product p-nitroaniline. This chromogenic product can be monitored continuously using a UV–Vis spectrophotometer set at 405 nm. The assay was tested with the type I MetAP from Escherichia coli (EcMetAP-I) and the type II MetAP from Pyrococcus furiosus (PfMetAP-II). Using l-Met-p-NA, the kinetic constants kcat and Km were determined for EcMetAP-I and PfMetAP-II and were compared with those obtained with a “standard” high-performance liquid chromatography (HPLC) discontinuous assay. The assay has also been used to determine the temperature dependence of the kinetic constant kcat for PfMetAP-II as well as to screen two novel pseudopeptide inhibitors of MetAPs. The results demonstrate that l-Met-p-NA provides a fast, convenient, and effective substrate for both type I and type II MetAPs and that this substrate can be used to quickly screen inhibitors of MetAPs

A first course in feedback, dynamics and control: findings from an online pilot survey for the IFAC community

Undergraduate students in many engineering programs around the world take only one control course. The IFAC Educational Committee has developed and piloted to a limited audience a comprehensive survey for the topics to be included in such course. This issue is relevant to both academia and industry. The paper discusses the initial findings related to the design of the survey as well as the responses of the participants. The findings will be used to refine the survey and distribute it in the near future to the global control community

Technical committee on control education

Presents information of the CS society Presents information of the CS society Technical Committee on Control Education

A survey of international views on a first course in systems and control for engineering undergraduates

This paper summarises the results of an international survey of academics and industrialists on what should be prioritised in the first, and often only, control course taken by engineering undergraduates. The results are made up both of quantitative data whereby respondents selected from a number of options, and also of qualitative data where respondents entered free comments. Reflections on the results and summaries of common trends are given to help readers consider how the curriculum in their own institutions might be updated and modified to meet modern requirements. At the time of writing the survey had around 500 respondents covering a good spread of nationalities, employment status and engineering disciplines

On an IFAC online pilot survey for a first course on control

The paper introduces aims and objectives of the IFAC survey for a first course on control and discusses some its basic features

Reduction of angular divergence of laser-driven ion beams during their acceleration and transport

Laser plasma physics is a field of big interest because of its implications in basic science, fast ignition, medicine (i.e. hadrontherapy), astrophysics, material science, particle acceleration etc. 100-MeV class protons accelerated from the interaction of a short laser pulse with a thin target have been demonstrated. With continuing development of laser technology, greater and greater energies are expected, therefore projects focusing on various applications are being formed, e.g. ELIMAIA (ELI Multidisciplinary Applications of laser-Ion Acceleration). One of the main characteristic and crucial disadvantage of ion beams accelerated by ultra-short intense laser pulses is their large divergence, not suitable for the most of applications. In this paper two ways how to decrease beam divergence are proposed. Firstly, impact of different design of targets on beam divergence is studied by using 2D Particlein-cell simulations (PIC). Namely, various types of targets include at foils, curved foil and foils with diverse microstructures. Obtained results show that well-designed microstructures, i.e. a hole in the center of the target, can produce proton beam with the lowest divergence. Moreover, the particle beam accelerated from a curved foil has lower divergence compared to the beam from a flat foil. Secondly, another proposed method for the divergence reduction is using of a magnetic solenoid. The trajectories of the laser accelerated particles passing through the solenoid are modeled in a simple Matlab program. Results from PIC simulations are used as input in the program. The divergence is controlled by optimizing the magnetic field inside the solenoid and installing an aperture in front of the device

Supplementary material for the article: Macháčková, K.; Chrudinová, M.; Radosavljević, J.; Potalitsyn, P.; Křížková, K.; Fábry, M.; Selicharová, I.; Collinsová, M.; Brzozowski, A. M.; Žáková, L.; et al. Converting Insulin-like Growth Factors 1 and 2 into High-Affinity Ligands for Insulin Receptor Isoform A by the Introduction of an Evolutionarily Divergent Mutation. Biochemistry 2018, 57 (16), 2373– 2382. https://doi.org/10.1021/acs.biochem.7b01260

Supplementary material for: [https://doi.org/10.1021/acs.biochem.7b01260]Related to published version: [http://cherry.chem.bg.ac.rs/handle/123456789/2135

Rational steering of insulin binding specificity by intra-chain chemical crosslinking

Insulin is a key hormone of human metabolism with major therapeutic importance for both types of diabetes. New insulin analogues with more physiological profiles and better glycemic control are needed, especially analogues that preferentially bind to the metabolic B-isoform of insulin receptor (IR-B). Here, we aimed to stabilize and modulate the receptor-compatible conformation of insulin by covalent intra-chain crosslinking within its B22-B30 segment, using the Cu I -catalyzed Huisgen 1,3-dipolar cycloaddition reaction of azides and alkynes. This approach resulted in 14 new, systematically crosslinked insulin analogues whose structures and functions were extensively characterized and correlated. One of the analogues, containing a B26-B29 triazole bridge, was highly active in binding to both IR isoforms, with a significant preference for IR-B. Our results demonstrate the potential of chemistry-driven modulation of insulin function, also shedding new light on the functional importance of hormones B-chain C-terminus for its IR-B specificity

Teaching control during the COVID-19 pandemic

This paper aims to analyze some different solutions that were adopted in control education activities during the pandemic. The authors of this paper are educators in the control education field from different countries on all the continents, who have developed a questionnaire with the idea of collecting data about the COVID-19 pandemic impact on the control education activities. The main objective is to study the diverse alternatives that were used worldwide to perform the online educational activities during that period, such as methodologies, tools, learning management systems (LMS), theoretical exercises, laboratory experiments, types of exams, simulators, software for online lecturing, etc. As a result, comparisons between pre-and during-pandemic educational resources and methods are performed, where useful ideas and discussions are given for the control education community.https://www.journals.elsevier.com/ifac-papersonlineElectrical, Electronic and Computer Engineerin

Engineering of Insulin Receptor Isoform-Selective Insulin Analogues

BACKGROUND: The insulin receptor (IR) exists in two isoforms, A and B, and the isoform expression pattern is tissue-specific. The C-terminus of the insulin B chain is important for receptor binding and has been shown to contact the IR just adjacent to the region where the A and B isoforms differ. The aim of this study was to investigate the importance of the C-terminus of the B chain in IR isoform binding in order to explore the possibility of engineering tissue-specific/liver-specific insulin analogues. METHODOLOGY/PRINCIPAL FINDINGS: Insulin analogue libraries were constructed by total amino acid scanning mutagenesis. The relative binding affinities for the A and B isoform of the IR were determined by competition assays using scintillation proximity assay technology. Structural information was obtained by X-ray crystallography. Introduction of B25A or B25N mutations resulted in analogues with a 2-fold preference for the B compared to the A isoform, whereas the opposite was observed with a B25Y substitution. An acidic amino acid residue at position B27 caused an additional 2-fold selective increase in affinity for the receptor B isoform for analogues bearing a B25N mutation. Furthermore, the combination of B25H with either B27D or B27E also resulted in B isoform-preferential analogues (2-fold preference) even though the corresponding single mutation analogues displayed no differences in relative isoform binding affinity. CONCLUSIONS/SIGNIFICANCE: We have discovered a new class of IR isoform-selective insulin analogues with 2-4-fold differences in relative binding affinities for either the A or the B isoform of the IR compared to human insulin. Our results demonstrate that a mutation at position B25 alone or in combination with a mutation at position B27 in the insulin molecule confers IR isoform selectivity. Isoform-preferential analogues may provide new opportunities for developing insulin analogues with improved clinical benefits