Current issue

The evolutionary pathway of diagnostic testing methods can be traced through the historical development and refinement of analytical tools. These tools are designed to identify sources, decipher mechanisms, and control processes inseparable from functional and morphological organization of living organisms under both normal and pathological conditions. In essence, this pathway represents a shift from subjective observation to objective formalization. The importance of this transition has been long recognized and is difficult to overestimate, as illustrated by the consequences of reckless human interference with nature. Furthermore, analysis of the global diagnostic systems market indicates significant growth: valued at USD 14.51 billion in 2021, it is forecast to reach USD 25.88 billion by 2029 (Data Bridge Market Research). The diversity of approaches to developing immunodiagnostic methods, as well as to biomedical analytics in general, is vast. Researchers continue to generate new methodologies. Accordingly, this review analyzes the evolutionary pathways of immunoassay techniques and tools that are actively applied in both laboratory and point-of-care testing.

This study aims to investigate the molecular mechanisms underlying the involvement of heat shock proteins 27 and 70 and ubiquitin in the oxidative modification of proteins, as well as the execution of dexamethasone-induced apoptosis in Jurkat tumor cells under inhibition of heat shock protein 27. It was assessed how 5-(5-ethyl-2-hydroxy-4-methoxyphenyl)-4-(4-methoxyphenyl)isoxazole at a final concentration of 0.1 µM and/or dexamethasone at a final concentration of 10 µM affected cytoplasmic exposure of phosphatidylserine followed by annexin V binding, the number of FasL and TNFα receptors, and the reduction of mitochondrial membrane potential in cells. Their effects were also examined with respect to the levels of OH• radicals, free sulfhydryl groups of cysteine in proteins, protein-bound glutathione, oxidized tryptophan residues, bi-tyrosine cross-links, carbonyl derivatives of proteins, ubiquitin, ubiquitin ligase, NFkB, Apaf-1, and heat shock proteins 27 and 70, as well as caspase-3 activity in Jurkat tumor cells.

The observed changes in the levels of heat shock proteins 27 and 70, ubiquitin, oxidative modifications of amino acid residues, and proteins were associated with the execution of apoptosis in Jurkat tumor cells. When exposed simultaneously to 5-(5-ethyl-2-hydroxy-4-methoxyphenyl)-4-(4-methoxyphenyl)-isoxazole and dexamethasone, Jurkat tumor cells exhibited correlations between the activation of irreversible oxidative modifications and the reduction of reversible oxidative modifications of proteins, associated with the execution of apoptosis involving TNFα and Fas receptors.

The findings indicate that heat shock proteins 27 and 70, together with ubiquitin, participate in both reversible and irreversible oxidative modifications of amino acid residues and proteins, as well as in the execution of dexamethasone-induced apoptosis in Jurkat tumor cells when heat shock protein 27 is inhibited.

Axial spondyloarthritis (AxSpA) is a chronic inflammatory disorder of the musculoskeletal system that belongs to the group of spondyloarthritis, characterized by the obligatory involvement of the sacroiliac joints and/or axial skeleton in the pathological process. In this study, we collected peripheral blood samples from 32 AxSpA patients and 29 healthy controls (НС), and investigated the regulatory T cell (Tregs) subsets by multicolor flow cytometry. We noticed that the relative numbers of ’naïve’ Tregs were decreased, while the relative and absolute numbers of central and effector memory Tregs (CM and EM were CD62L + CD45RA – and CD62L – CD45RA –, respectively) were elevated in patients with AxSpA. Next, in AxSpA patients CXCR5 – CXCR3 + CCR6 – Tr1 cells were increased both in relative and absolute frequencies. The absolute numbers of CXCR5 – CXCR3 – CCR6 – Tr2, CXCR5 – CXCR3 – CCR6 + Tr17, and CXCR5–CXCR3+CCR6+ Tr17.1 were also elevated in patients with AxSpA vs. HC group. Finally, in patients with AxSpA Tr1 were increased exclusively within CM Tregs, while Tr2 cells were elevated both within CM and EM Treg subsets. Tr17 cells were decreased both within CM and EM Treg subsets. Interestingly, we found no significant differences in CXCR5+ follicular Tregs between the groups. Further investigations of circulating Tregs will contribute to find new targets and therapies in AxSpA.

Infectious keratitis and corneal ulcers are inflammatory diseases of the corneal tissue caused by various types of bacteria, characterized by both acute and chronic course, and also manifested in the form of progressive ulceration and rapidly progressing purulent infection of any part of the corneal tissue. Bacterial corneal ulcers are one of the most severe eye pathologies due to the risk of corneal perforation and infection with the development of endophthalmitis. The aim of the study was to conduct molecular genetic studies to determine bacterial pathogens spectrum in various biological material of patients with keratitis and corneal ulcers. Pathogenic microorganisms were detected by real-time PCR with qualitative and quantitative formats. Etiologically significant microorganisms in corneal ulcers are Streptococcus species and Staphylococcus species, which were identified in the ophthalmological biological material of patients with own cornea ulcers in 80.00 ± 8.20% and 70.00 ± 7.56% of cases, respectively, and in 78.57 ± 8.36% and 64.29 ± 7.65% of cases with corneal transplant ulcers; in keratitis, Streptococcus species DNA was identified in 57.89 ± 7.18%, Staphylococcus species — 42.11 ± 6.22%. Significantly (p < 0.05) higher values of DNA concentration (106 GE/ml) were found in upper respiratory tract epithelial cells scrapings in comparison with eye biological material (104—105 GE/ml). The upper respiratory tract should be considered as a possible source of eye infection, which is confirmed by quantitative data on determining DNA concentrations.

The outbreak of the COVID-19 pandemic, announced in March 2020, caused unprecedented alarm among people around the world. At that time, few people could have predicted that the virus was capable of becoming chronic, but soon the term "cystic syndrome" was widely recognized in the scientific and medical community. It was recorded that by 2023, the number of COVID-19 cases had significantly decreased compared to the peak values of 2020—2022. However, after patients suffered from COVID-19, many began to face a deterioration in their health, the development of complications and the reactivation of chronic infections such as herpes virus and papillomavirus infections. Herpes viruses are able to enter a latent, chronic state in the cells of the nervous and immune systems, waiting for a trigger situation for reactivation, in the form of a weakening of the general immune system. The complement system and neutrophils are links of innate immunity and interact with each other. However, data on the functioning of the complement system and the phagocytic activity of neutrophils in postcovoid syndrome, especially in combination with chronic herpesvirus infection, remain limited. These reasons highlight the impor­tance of the current research and encourage further study of the field.

This paper for the first time presents an efficient method for the synthesis of 3,4-dihydropyrimidin-2(1H)-ones (3,4-DHPMs) bearing aliphatic and cyclopropane-containing aldehyde residues, using europium(III) chloride hexahydrate as a catalyst in the Biginelli multicomponent reaction (MCR). A high permeability through the phospholipid bilayer and the potential for passive diffusion facilitating participation in in­tracellular interactions were determined. A preliminary eva­luation of the antifungal properties of the obtained compounds against a range of fungal species was carried out using in silico
methods. Experiments on the effect of 3,4-dihydropyrimi­dinones on the growth of Yarrowia lipolytica yeast showed no acute toxicity of the tested compounds within the micromolar concentration range.