Thioflavin T (ThT) assay is extensively used for studying fibrillation kinetics in vitro. However, the differences in the time course of ThT fluorescence intensity and lifetime and other physical parameters of the system, such as particle size distribution, raise questions about the correct interpretation of the aggregation kinetics. In this work, we focused on the investigation of the mechanisms, which underlay the difference in sensitivity of ThT fluorescence intensity and lifetime to the formation of protein aggregates during fibrillation by the example of insulin and during binding to globular proteins. The assessment of aggregate sizes and heterogeneity was performed using dynamic light scattering (DLS) and nanoparticle tracking analysis (NTA). Using the sub-nanosecond resolution measurements, it was shown that the ThT lifetime is sensitive to the appearance of as much as a few percent of ThT bound to the high-affinity sites that occur simultaneously with an abrupt increase of the average particle size, particles concentration, and size heterogeneity. The discrepancy between ThT fluorescence intensity and a lifetime can be explained as the consequence of a ThT molecule fraction with ultrafast decay and weak fluorescence. These ThT molecules can only be detected using time-resolved fluorescence measurements in the sub-picosecond time domain. The presence of a bound ThT subpopulation with similar photophysical properties was also demonstrated for globular proteins that were attributed to non-specifically bound ThT molecules with a non-rigid microenvironment.

The impaired balance of water distribution in the human organism accompanies numerous pathological states, and measuring water content in tissues is required for biomedical diagnostics. For instance, during heart failure,¹ physical examination using the “pitting scale” and weighing the patient remains the most useful approaches for assessing edematous syndrome; however, these methods are subjective and insufficient.² Assessment of water and lipids content in breast cancer tissues can provide important information about the characteristics of a tumor,³ as the high water content in a tumor indicates edema and increased cellularity.^4,5 Likewise, the measurement of hydration in different skin layers is interesting for dermatology and cosmetology:⁶ for instance, water content in the dermis is a marker of changes in the elastic properties of the skin,⁷ and hydration of the stratum corneum is indicative of its barrier function.⁸ Hence, the necessity of water content assessment stimulates the development of appropriate techniques, including those based on optical spectroscopy.⁹

A reaction of acyl chlorides derived from 1,10-phenanthroline-2,9-dicarboxylic acids with piperazine allows the preparation of the corresponding 24-membered macrocycles in good yield. The structural and spectral properties of these new macrocyclic ligands were thoroughly investigated, revealing promising coordination properties towards f-elements (Am, Eu). It was shown that the prepared ligands can be used for selective extraction of Am(III) from alkaline–carbonate media in presence of Eu(III) with an SF_Am/Eu up to 40. Their extraction efficiency is higher than calixarene-type extraction of the Am(III) and Eu(III) pair. Composition of macrocycle–metal complex with Eu(III) was investigated by luminescence and UV-vis spectroscopy. The possibility of such ligands to form complexes of L:Eu = 1:2 stoichiometry is revealed.

Edema, i.e., fluid accumulation in the interstitial space, accompanies numerous pathological states of the human organism, including heart failure (HF), inflammatory response, and lymphedema. Nevertheless, techniques for quantitative assessment of the edema’s severity and dynamics are absent in clinical practice, and the analysis is mainly limited to physical examination. This fact stimulates the development of novel methods for fast and reliable diagnostics of fluid retention in tissues. In this work, we focused on the possibilities of two microscopic techniques, nailfold video capillaroscopy (NVC) and confocal laser scanning microscopy (CLSM), in the assessment of the short-term and long-term cutaneous edema. We showed that for the patients with HF, morphological parameters obtained by NVC—namely, the apical diameter of capillaries and the size of the perivascular zone—indicate long-term edema. On the other hand, for healthy volunteers, the application of two models of short-term edema, venous occlusion, and histamine treatment of the skin, did not reveal notable changes in the capillary parameters. However, a significant reduction of the NVC image sharpness was observed in this case, which was suggested to be due to water accumulation in the epidermis. To verify these findings, we made use of CLSM, which provides the skin structure with cellular resolution. It was observed that for the histamine-treated skin, the areas of the dermal papillae become hyporefractive, leading to the loss of contrast and the lower visibility of capillaries. Similar effect was observed for patients undergoing infusion therapy. Collectively, our results reveal the parameters can be used for pericapillary edema assessment using the NVC and CLSM, and paves the way for their application in a clinical set-up.

Osteoarthritis is a degenerative disease that affects articular cartilage, leading to changes on the macro and micro levels of this multi-component tissue. Understanding the processes underlying this pathology plays an important role in planning the following management tactics. Timely detection of the knee joint degradation at the level of tissue changes can prevent its progressive damage due to the early beginning of appropriate treatment. This study aimed to provide an overview of the current level of knowledge about the composition of cartilage and menisci using a wide range of different diagnostic methods. A systematic review of the literature published from 1978 to 2023 was conducted. Original studies of the knee joint cartilage (articular and meniscus) research, reporting content composition and mechanical properties, were included. Studies of the non-knee joint cartilage, tissue research other than cartilage and meniscus, or reporting treatment outcomes were excluded (n = 111). Thirty-one papers were included in this review, which reported on the composition of animal and human cartilage (articular and meniscus). The most frequently investigated parameters were quantitative proteoglycan determination and hydration level of the cartilage. Cartilage and meniscus degeneration, i.e., reduced collagen and proteoglycan content, reduced mechanical properties, and increased hydration level, was shown in every article about osteoarthritis. Among all diagnostic methods, laboratory methods (biochemical and histological analysis) are the most frequently used, compared to the instrumental ones (spectroscopy, MRI, and CT). At the same time, spectroscopy takes the lead and becomes the most common approach for determining cartilage composition (collagen and proteoglycans content). Graphical Abstract