Azodicarboxylate esters are common reagents in organic synthesis laboratories due to their utility in the Mitsunobu reaction. They can also be regarded as possible starting compounds for C–H functionalization, which up until now has been mainly achieved by transition-metal-catalyzed reactions. We have developed a novel reaction involving the quantitative coupling of 8-hydroxyquinoline or phenol with azodicarboxylate esters. The functionalization proceeds under mild base-catalyzed conditions selectively, and either the ortho-position of 8-hydroxyquinoline or para-position of the phenol/naphthol is involved in the reaction. This type of transformation can be considered as “hydrazo coupling” (by analogy with azo coupling). Herein, we discuss a plausible mechanism for this catalyzed substitution, backing up our findings with deuterium NMR experiments and by varying the starting compounds and bases. Using Boc-NN-Boc as a substrate, we have developed the convenient and efficient synthesis of (8-hydroxyquinolin-7-yl)hydrazines, as well as demonstrating a new stereoselective route for the synthesis of medicinally important 4-hydroxyphenylhydrazine for laboratory use, which almost doubles the yield of the common industrial process and reduces the number of synthetic steps. A new “one-pot” procedure for the synthesis of aromatic 8-hydroxyquinolin-7-yl hydrazones was applied.

Aromatic hydrazones of 7-hydrazino-8-hydroxyquinoline were studied as anti-HIV and antibacterial compounds. A set of the compounds with different aromatic moieties bearing electron-donating and electron-withdrawing substituents has been selected and obtained via the hydrazo coupling and “one-pot” click reaction with aldehydes. The compounds possess activity against both bacterial and fungal targets. Cellular Ku70-inhibiting activity has been found for the series, opening a new class of inhibitors for potential anti-HIV treatment. The compounds display anti-HIV activity in infected cells at submicromolar concentrations. Their low solubility can be overcome by incorporation in water-soluble neutral polyalginate microgels (33% wt load of the compound).

To date, the problem of mastitis in cattle remains relevant for both the industrial sector and scientific research. Despite numerous active investigations, the causes of this disease have not been fully established. It is postulated that several factors may be involved, such as bacterial pathogens, animal husbandry practices, and weather and climatic conditions. In this study, we selected cows from farms in Yakutia to investigate microbial isolates present in the milk of cows affected by mastitis and treated with antibiotics. Five identified Staphylococcus aureus isolates were investigated using whole-genome sequencing (Illumina sequencing and nanopore sequencing), followed by analysis of virulence factors in the genomes and cultural properties of the isolates. The profile of S. aureus virulence genes (exotoxins, cytotoxins, superantigen-like proteins, adhesins) was identified via WGS. Hemolysin gene (hla) was detected in all isolates. An investigation of the cultural properties of the isolates, specifically through hemolysis of rabbit erythrocytes and Western blot analysis of the culture liquid of S. aureus, revealed different expression levels of alpha-hemolysin among the strains. One isolate (17-21) exhibited the highest secretion level of about 320 ± 37 ng, both in the hemolysis test and immunoblotting assay. An investigation of the isolates’ antibiotic resistance showed that all isolates exhibited multidrug resistance, as confirmed by the presence of antibiotic resistance genes in these isolates. One isolate (7-7) exhibited the broadest range of phenotypic resistance and was resistant to all tested antibiotics (except clindamycin). Phylogenetic analysis suggested that the evolution of these isolates occurred independently in their respective ecological niches, although their transfer from cattle to humans, and vice versa, is possible. Isolates 7-7, 18-22, 33-40, and 35-42 are most typical to Yakutian cattle, while isolate 17-21 might have been introduced from a different region. To the best of our knowledge, this is the first in-depth study into a range of S. aureus isolates associated with mastitis infection in Yakutian cattle.

The generation of induced pluripotent stem cells (iPSCs) from peripheral blood T-lymphocytes offers a promising alternative to skin fibroblasts. This approach combines minimally invasive sample collection with rapid isolation of enriched target cell population and provides a lower baseline mutational burden, as T-lymphocytes are shielded from chronic ultraviolet radiation – a major driver of somatic mutations in skin fibroblasts. Objective verification of monoclonality of the resulting iPSC lines is possible due to V(D)J rearrangements in T-cell receptor (TCR) genes, which are formed as a result of somatic recombination in the thymus during the natural maturation of T-lymphocytes. These rearrangements remain unchanged during reprogramming and serve as a unique marker, allowing for reliable identification of monoclonal lines and exclusion of contaminated or polyclonal lines. Reliable verification of the clonal origin of iPSCs can be crucial in experiments that place high demands on genetic homogeneity. This work is dedicated to the generation and comprehensive characterization of monoclonal iPSC lines derived from CD3+ T-lymphocytes of a healthy donor using episomal reprogramming. The resulting lines, RCPCMi014-A (PBM022E5) and RCPCMi014-B (PBM022E7), meet the accepted quality criteria for iPSCs: they demonstrate expression of key pluripotency markers (OCT4, SOX2, SSEA-4, TRA-1-81), the ability to differentiate into derivatives of all three germ layers, and possess a normal karyotype. Both lines showed a high-efficiency capacity to differentiate into definitive endoderm, as assessed by CXCR4 expression, highlighting their potential for developing protocols to generate pancreatic β-cells. The obtained iPSC lines can be used for fundamental research into the mechanisms of pluripotency and differentiation, as well as for creating isogenic iPSC lines with introduced mutations for modeling rare hereditary diseases.