Mitogen-activated protein kinases, ERK1/2 (MAPK3/1), play a key role in the regulation of cell growth, differentiation, and apoptosis. We have previously presented evidence proving that activation of the ERK1/2 axis in cancer cells following the administration of therapeutics leads to the overexpression of growth factor receptors and drug resistance. Recently, we have proposed a new bioinformatic technique that enables direct construction of interactome network-based molecular pathways for gene products of interest, as well as quantitation of their activation levels using high-throughput gene expression data. In this study, we, for the first time, algorithmically constructed ERK1/2 molecular pathways and investigated how their activation levels (PALs) affect survival and responsiveness to targeted drugs at the pan-cancer level based on transcriptomic data. We examined a total of 11 287 human tumor profiles from 31 types of cancer, drawn from 53 of our previously published and other literature datasets, looking at patient survival and clinical response to 29 chemo- and targeted therapy regimens. We found that activation of the ERK1/2 pathways has different prognostic significance depending on cancer type. In glioblastoma, sarcoma, lung, kidney, bladder, gastric, colon, and several other cancer types, ERK pathway activation was associated with worse survival. In contrast, the same phenomenon was associated with a better chance of survival in HER2+, luminal A and luminal B breast cancer, and uterine corpus cancer. These trends were consistent with treatment response analysis. At the same time, we found significantly worse associations with the expression levels of individual MAPK1 and MAPK3 genes: hence, ERK1/2 pathway activation levels can be considered putative biomarkers for predicting clinical outcomes and selecting new personalized treatment strategies, such as the use of MAPK inhibitors.

Antibiotic resistance threatens global healthcare. In clinical practice, conventional antibiotics are becoming gradually less effective. Moreover, the introduction of new antimicrobial agents into clinical practice leads to the emergence of resistant pathogenic strains within just a few years. Hence, the development of platforms for massive creation and screening of new antimicrobial agents is of particular importance. Massive parallel screening will greatly reduce the time required to identify the most promising drug candidates. Meanwhile, DNA-encoded antimicrobial agents offer unique opportunities for the high-throughput development of new antibiotics. Here, the yeast Pichia pastoris was engineered to produce a panel of antimicrobial peptides (AMPs), followed by high-throughput screening of AMP producers that inhibit bacterial growth in situ. Yeast clones producing thanatin and protegrin-1 exhibited the highest level of antimicrobial activity among the panel of AMPs under investigation. The production level of recombinant thanatin was significantly higher than that of protegrin-1, which correlates with its low toxicity. The designed technique of massive assessment of the activity of DNA-encoded antimicrobial agents enables the identification of drug candidates with an increased therapeutic index. Further development of methods for a rational design of artificial diversity in AMPs, followed by deep functional profiling of antimicrobial activity, will yield new AMPs with improved therapeutic characteristics.

Immunoregulatory enzymes, which function both as biological catalysts and regulatory elements, play a crucial role in controlling immune responses. Dysfunction of these proteins can contribute to various pathological conditions, such as the suppression of antitumor immunity or impairment of anti-infectious immune responses. This review discusses the most extensively studied immunoregulatory enzymes, including indoleamine 2,3-dioxygenase 1, arginase 1, inducible nitric oxide synthase, glyceraldehyde-3-phosphate dehydrogenase, and ectonucleoside triphosphate diphosphohydrolase 1. Their classification is provided, along with an analysis of the distinctive characteristics inherent to this group of enzymes. Additionally, new directions for the medical application of immunoregulatory enzymes are explored.

Neurodegenerative disorders classified as synucleinopathies (Parkinson’sdisease, dementia with Lewy bodies, and multiple-system atrophy) arecharacterized by the accumulation of aberrant α-synuclein aggregates inneurons and glial cells. These diseases manifest clinically several years afterthe initial formation of pathological protein aggregates in the brain, makingearly and accurate diagnosis challenging. In recent years, a new method, whichis based on real-time quaking-induced conversion (RT-QuIC) of α-synuclein,has been developed and validated. This technology holds great promise as apowerful diagnostic tool for the early and precise identification ofsynucleinopathies, potentially opening new horizons in the study ofneurodegenerative diseases. RT-QuIC detects misfolded α-synucleinaggregates in human physiological fluids by introducing an excess ofrecombinant α-synuclein, which undergoes conformational conversion in anexponential, prion-like manner. The production of high-quality recombinantα-synuclein is a critical step in the effective application of thismethod, as protein purity significantly affects the sensitivity and specificityof the assay — key factors in its diagnostic utility. Using a three-stepchromatographic purification protocol, we produced recombinant monomericα-synuclein with a purity exceeding 97% from the periplasmic fraction ofbacterial cells. While higher purity increases the assay duration, it alsoreduces the background signal and permits extended incubation times, which areessential for reliably detecting synucleinopathies with weak RT-QuIC responses,such as the cerebellar subtype of multiple-system atrophy. The data presentedsupport the conclusion that optimized components of the RT-QuIC system willenable an accurate diagnosis of neurodegenerative synucleinopathies.

A patient with an immunophenotype characteristic of B-cell acute lymphoblasticleukemia (B-ALL) was found to carry the chromosomal translocationt(9;22)(q34;q11), or Philadelphia (Ph) chromosome and less common variant ofthe chimeric oncogene BCR::ABL/p210. No additional mutations in the BCR::ABLgene, including point mutations, insertions, or deletions, were identified inthe disease onset characterized by elevated blast cell (77.6%) and leukocyte(48×109/L) counts. Ph+ALL-2012m chemotherapy with imatinib (600 mg) andtwo consolidation phases resulted in complete hematologic remission and aprofound molecular response. However, six months later, the patient hadrelapsed (blasts: 15%, BCR::ABL/p210: 105%). Three weeks after the initiationof dasatinib therapy (100 mg), the number of blasts had decreased to 4.8%,while the expression level of BCR::ABL/p210 had dropped to 11.8%. Sangersequencing identified two mutations in the BCR::ABL oncogene; namely, the pointmutation F317L and a new insertion of nine nucleotides previously not detected.In the latter case, the amino acid lysine at position 294 was replaced by fournew amino acid residues: K294SPSQ. Therapy with bosutinib and inotuzumab led tothe disappearance of one leukemia clone with the F317L mutation, but thepresence of another clone carrying a nine-nucleotide insertion was observed.The switch to ponatinib+blinatumomab chemotherapy was effective, resulting inthe disappearance of the insertion. Allogeneic hematopoietic stem celltransplantation (allo-HSCT) from an available HLA-matched unrelated donorresulted in complete clinical and hematologic remission, including a completemolecular response. Six months after allo-HSCT, minimal residual diseasemonitoring showed maintenance of complete remission.

The efficiency of dairy product fermentation directly depends on the propertiesof the lactic acid bacteria used, particularly on their metabolic activity andresistance to bacteriophages. Therefore, an understanding of the relationshipsbetween the genetic and phenotypic traits of industrial strains is of elevatedimportance. In this study, we comprehensively analyzed five Lactococcus strainswidely used in the Russian dairy industry, combining whole-genome sequencingwith phenotypic profiling. Despite the fact of genetic similarity among four ofthe L. lactis strains, we still identified significant differences intheir metabolic activity. Comparative structural analysis of previouslypublished genomes of 337 L. lactis and 147 L. cremoris strainsrevealed species-specific features of the lactose metabolism; in particular,the absence of the lacZ gene in L. cremoris. Notably, prophages were foundin three of the studied strains, which was in correlation with their reducedacidification activity. L. lactis FNCPS 51n and 73n strains displayedresistance to all 50 tested bacteriophages, which may be associated with thepresence of the AbiB abortive infection system. These findings underscore theimportance of integrating genomic and phenotypic analyses when selectingefficient and phage-resistant Lactococcus starters in the dairy industry.

Boron neutron capture therapy (BNCT) is a rapidly developing field of radiationtherapy for cancer that is based on the accumulation of the radiosensitive 10Bisotope in cancer cells, followed by tumor irradiation with thermal neutrons.Widespread use of BNCT in clinical practice remains limited because of the pooraccumulation of boron-containing (10B) drugs in the tumor or their hightoxicity to the body. This study focuses on the engineering of tumor-specificliposomes loaded with 4-L-boronophenylalanine (4-L-10BPA) for application inboron neutron capture therapy. According to the spectrophotometry and ICP-massspectroscopy data, the 4-L-10BPA-to-liposome molar ratio is ~ 120,000.Liposomal targeting of human epidermal growth factor receptor 2 (HER2) wasdetermined by HER2-specific designed ankyrin repeat protein (DARPin)_9-29 onthe outer surface of liposomes. DARPin-modified liposomes were found to bind toHER2-overexpressing cells and be effectively internalized into the cytoplasm.The ability of DARPin-functionalized liposomes to precision-deliver largequantities of 4-L-10BPA into cancer cells may open up new prospects for BNCT.

OrthopoxVac, a fourth-generation smallpox vaccine, was the first of its kindregistered worldwide in 2022, and it has been shown to be both safe and toinduce only a mild reaction. A six-month clinical study confirmed itsimmunogenicity as compared to the first-generation live smallpox vaccine. Ourstudy aimed to determine the levels of specific humoral and T-cell immuneresponses in volunteers following intradermal OrthopoxVac vaccineadministration either in a single dose of 107 PoFU or in two doses of 106 PoFU,at 1.5, 3, and 5 years after initial vaccination. Following the immunization ofvolunteers with the OrthopoxVac vaccine at a dosage of 107 PoFU, the T-helperresponse remained at a relatively high level for three years, before itsignificantly dropped. Administration of the same vaccine twice at a dose of106 PoFU resulted in a considerable decrease in the level of T-helpers, after1.5 years. Additionally, some patients exhibited a reduction in viralneutralizing antibody (VNA) titers after 1.5 years of OrthopoxVac vaccineadministration. When OrthopoxVac was administered at a dosage of 107 PoFU, nosubstantial differences were noted between groups at the 1.5-, 3-, and 5-yearmarks. In contrast, in the groups receiving two doses of 106 PoFU, VNA titersshowed a significant reduction after 1.5 years. These findings indicate that asingle intradermal dose of 107 PoFU of the OrthopoxVac vaccine elicits asignificant and lasting immune response involving both antibodies and T-cellsfor a minimum of three years.

Cancer is one of the leading causes of death worldwide. Traditional cancer treatments include surgery, radiotherapy, and chemotherapy, as well as combinations of these treatments. Despite significant advances in these fields, the search for innovative ways to treat malignant tumors, including the application of oncolytic viruses, remains relevant. One such virus is the vesicular stomatitis virus (VSV), which possess a number of useful oncolytic properties. However, VSV-based drugs are still in their infancy and are yet to be approved for clinical use. This review discusses the mechanisms of oncogenesis, the antiviral response of tumor and normal cells, and markers of tumor cell resistance to VSV virotherapy. In addition, it examines methods for producing and arming recombinant VSV and provides examples of clinical trials. The data presented will allow better assessment of the prospects of using VSV as an oncolytic.

The increasing resistance of microorganisms to antibiotics makes it a necessity that we search for new antimicrobial agents. Due to their genetically encoded nature, peptides are promising candidates for new antimicrobial drugs. Lantipeptide andalusicin exhibits significant antimicrobial activity against Gram-positive bacteria, making it a promising scaffold for the development of DNA-encoded libraries of lantibiotics. In this study, the modification reaction of andalusicin by class III lanthionine synthetase AncKC was reconstructed in a heterologous Escherichia coli system. The results obtained open possibilities for creating novel peptide- based antimicrobial agents.