Summary Rapid identification of pathogen and its resistance to antimicrobial drugs, and subsequent appropriate antimicrobial treatment are essential for correct patient outcomes. Conventional detection methods of bacterial resistance, such as disc diffusion, broth microdilution and automated instruments, are constantly widely used and primarily standardized. Nevertheless, the results cannot be obtained earlier than 48 h after receiving a sample, which may lead to prolonged use or overuse of broad‐spectrum antibiotics. Hence, there is a drive to develop and introduce novel, faster, standardized, sensitive and specific methods with reliable results into routine microbiological laboratory practice. Recently developed matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry (MALDI‐TOF MS) has been introduced in recent years into laboratory practice, and methods based on microfluidics and microdroplets might be introduced in the near future. This review is focused on the methods and instruments in use both currently and in the foreseeable future, applicable to determine antimicrobial efficacy in clinical microbiology laboratories.

Due to its sensitivity and productivity, bottom-up proteomics based on liquid chromatography−mass spectrometry (LC−MS) has become the core approach in the field. The de facto standard LC−MS platform for proteomics operates at sub-μL/min flow rates, and nanospray is required for efficiently introducing peptides into a mass spectrometer. Although this is almost a "dogma", this view is being reconsidered in light of developments in highly efficient chromatographic columns, and especially with the introduction of exceptionally sensitive MS instruments. Although conventional-flow LC−MS platforms have recently penetrated targeted proteomics successfully, their possibilities in discovery-oriented proteomics have not yet been thoroughly explored. Our objective was to determine what are the extra costs and what optimization and adjustments to a conventional-flow LC−MS system must be undertaken to identify a comparable number of proteins as can be identified on a nanoLC−MS system. We demonstrate that the amount of a complex tryptic digest needed for comparable proteome coverage can be roughly 5-fold greater, providing the column dimensions are properly chosen, extra-column peak dispersion is minimized, column temperature and flow rate are set to levels appropriate for peptide separation, and the composition of mobile phases is fine-tuned. Indeed, we identified 2 835 proteins from 2 μg of HeLa cells tryptic digest separated during a 60 min gradient at 68 μL/min on a 1.0 mm × 250 mm column held at 55 °C and using an aqua−acetonitrile mobile phases containing 0.1% formic acid, 0.4% acetic acid, and 3% dimethyl sulfoxide. Our results document that conventional-flow LC−MS is an attractive alternative for bottom-up exploratory proteomics.

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Alzheimer's disease (AD) is a major public health problem, which is due to its increasing prevalence and lack of effective therapy or diagnostics. The complexity of the AD pathomechanism requires complex treatment, e.g. multifunctional ligands targeting both the causes and symptoms of the disease. Here, we present new multitarget-directed ligands combining pharmacophore fragments that provide a blockade of serotonin 5-HT receptors, acetyl/butyrylcholinesterase inhibition, and amyloid β antiaggregation activity. Compound 12 has displayed balanced activity as an antagonist of 5-HT receptors ( K = 18 nM) and noncompetitive inhibitor of cholinesterases (IC = 14 nM, IC = 22 nM). In further in vitro studies, compound 12 has shown amyloid β antiaggregation activity (IC = 1.27 μM) and ability to permeate through the blood-brain barrier. The presented findings may provide an excellent starting point for further studies and facilitate efforts to develop new effective anti-AD therapy.