Antibiotic-resistant Pseudomonas aeruginosa presents a critical global health challenge, particularly in hospital-acquired infections. Bacteriophages offer a promising therapeutic avenue due to their ability to target and lyse resistant strains. This study characterizes Pseudomonas phage Banzai, a newly isolated Pbunavirus (family Lindbergviridae) with lytic activity against multiple P. aeruginosa isolates, including multidrug-resistant strains. Genomic analysis revealed a 66,189 bp genome, lacking antibiotic resistance or virulence factors, and suggested a headful packaging mechanism and the presence of a bidirectional component in the replication. In vivo experiments using Galleria mellonella showed therapeutic potential, significantly improving larval survival (87% at 24 h). Host range analysis revealed activity against 13 of 30 P. aeruginosa isolates, including members of O1, O3, O5 and O6 in silico predicted serogroups. Phylogenomic analyses place phage Banzai within the genus Pbunavirus, sharing 94.8% intergenomic similarity with its closest relatives, supporting its classification as a novel species. These findings highlight phage Banzai as a potential candidate for phage therapy, demonstrating genomic stability, a strictly lytic lifestyle, and in vivo efficacy.

Current scRNA-seq studies of solid tissues mostly rely on enzymatic dissociation of fresh samples or the fallback on nuclei isolation from frozen or partially fixed samples. However, due to the complex tissue organization or cell fragility, it could be challenging to apply these approaches to the sensitive endocrine tissues. That is, dissociating intact cells from such problematic fresh-frozen samples routinely collected by biobanks remains challenging. In this study, we adapted the acetic-methanol dissociation method – ACME High Salt (ACME HS) to effectively isolate intact single cells from fresh-frozen endocrine tumor samples, including adrenal gland neoplasms, thyroid carcinomas, and pituitary neuroendocrine tumors. We compared the ability of enzymatic, ACME HS, and nuclear isolation methods to preserve the integrity of major cell types and gene expression across 41 tissue samples of different origins. We demonstrated that ACME HS simultaneously dissociates and fixes cells, thus preserving morphology and a high RNA integrity number in problematic cell types. This finding renders the ACME HS dissociation method a valuable alternative in scRNA-seq protocols for challenging tissues where obtaining live cell suspension is difficult or impossible.

Background: The ACE Y215C mutation is a common, functionally damaging missense variant (~1.5% allele frequency) associated with reduced plasma ACE levels and increased Alzheimer’s disease (AD) risk. In CHO and HEK cell models, this mutation caused a ~3–6-fold decrease in ACE surface expression, soluble ACE levels, and ACE enzymatic activity compared to those of wild-type ACE. Methods: Circulating ACE levels and activity were measured in EDTA plasma obtained from 84 carriers of the ACE Y215C mutation using a set of mAbs to the ACE. The mAbs 5B3/1G12 binding ratio was revealed as a sensitive marker for the circulating Y215C ACE mutant. Whole-exome and whole-genome sequencing (WES/WGS) were performed to identify genetic variants potentially modifying circulating ACE levels. In parallel, published sequencing and proteomic data from 35,559 Icelanders participants were analyzed to identify genes influencing ACE shedding. Sequence comparison was performed between carriers with elevated and reduced ACE concentrations to identify the potential protective variants that may compensate for decreased ACE levels due to the Y215C mutation itself. Results: Most carriers of the Y215C ACE mutation demonstrated significantly decreased ACE levels (median is 62% of control ACE levels). However, substantial inter-individual variability was observed in plasma ACE activity among carriers. Comparative sequencing analysis revealed 9648 variants unique to individuals with elevated ACE, mapping to 5779 protein-coding genes and enriched for pathways related to intracellular and transmembrane transport. Conclusions: The presence of the damaging ACE mutation Y215C does not invariably result in low plasma ACE or, likely, elevated AD risk. Therefore, combined blood ACE phenotyping and whole-exome sequencing are recommended to more accurately assess ACE-related AD susceptibility in mutation carriers.

Background: Personalized pharmacotherapy requires systematic consideration of genetic factors influencing drug efficacy and safety. The accumulation of large-scale whole-exome sequencing (WES) resources provides an opportunity to assess population frequencies of clinically significant pharmacogenetic variants; however, the applicability of exome-based pharmacogenomics across populations, particularly those underrepresented in existing reference datasets, requires further evaluation. Materials and methods: A retrospective analysis of 6102 anonymized sequencing datasets obtained between 2020 and 2025 was performed using the DNBSEQ-G400 (MGI) platform and Agilent SureSelect Human All Exon v6/v7/v8 enrichment kits. SNV and indel detection, CNV analysis, high-resolution HLA typing, and diplotype assignment for key pharmacogenes were conducted. Pharmacogenomic annotations were derived from ClinPGx (formerly PharmGKB) (levels of evidence 1 A–2B), CPIC, and PharmVar. Haplotype phasing (SHAPEIT5), genotype imputation (IMPUTE5), and phased linkage disequilibrium analysis were performed using a reference panel of 814 Russian whole-genome sequences generated with the same bioinformatic workflow to assess the feasibility of reconstructing clinically relevant non-coding pharmacogenetic variants not captured by WES. Results: WES reliably detected 33 of 34 Very Important Pharmacogenes (VIPs), allowing determination of allele frequencies, metabolizer statuses for 13 VIPs, and HLA diversity. The highest allelic and phenotypic variability was observed in CYP2D6 , CYP2C19 , and CYP2B6 . A total of 663 ClinPGx annotations were identified, predominantly related to drug metabolism (50.38%) and toxicity (29.56%), involving psychotropic drugs, anticoagulants, statins, opioid analgesics, antineoplastic agents, and immunosuppressants. However, at least 32 drugs require assessment of non-coding pharmacogenetic variants or accurate CYP2D6 copy number determination – both beyond standard WES capability. Phased haplotype analysis demonstrated that haplotype-based imputation was feasible for only a limited subset of clinically relevant non-coding pharmacogenetic variants, whereas the majority lacked informative exonic proxies required for accurate inference from WES data. Conclusion: These findings represent one of the largest reference resource to date of pharmacogenetically significant variant and HLA allele frequencies in the Russian population. The results confirm WES as a robust and scalable platform for population-level pharmacogenomic screening and broader clinical genomic applications, while demonstrating that haplotype-based imputation cannot fully overcome the intrinsic limitations of exome sequencing for comprehensive pharmacogenomic profiling. Selection of genomic testing strategies should balance diagnostic completeness, analytical complexity, cost, scalability, and long-term potential for reanalysis.

Acne (acne vulgaris) is an extremely common dermatosis with specific clinical manifestations on the skin in the form of comedones (open and closed), inflammatory elements (papules, pustules and nodules), scars and post-inflammatory pigmentation, which, according to various data, affects more than 90% of adolescents and causes scarring on average in 20% of them. Genetic predisposition occupies an important place among a number of factors that can influence the disease course, as evidenced by the results of numerous studies. Objective. To determine the SNP of TNF (rs1800629) and TGF-β2 (rs1159268) genes significance in the development of moderate acne. Material and methods. The clinical anamnestic and molecular genetic methods were used. The research involved 204 patients aged between 17 and 46 years. The study group consisted of 102 patients with moderate acne (45 (44.1%) men and 57 (55.9%) women) at the age from 17 to 44 years, the control group included 102 conditionally healthy subjects (41 (40.2%) men, 61 (59.8%) women) aged between 18 and 46 years. The association of polymorphisms of GG TNF (rs1800629) and AA TGF-β2 (rs1159268), which increase the risk of developing disease by 2.5 and 3 times, respectively, with the development of moderate acne has been identified. These data allow to use the named indicators as risk markers for the development of moderate acne.

Background/Objectives: In recent years, research has highlighted the heterogeneity of corticotroph pituitary neuroendocrine tumors (CPitNETs), which are linked to Cushing’s disease. Purpose of the study is to determine stable postgenomic markers depending on maximum diameter on MRI (without visualization of the pituitary tumor on MRI/less than 6 mm (group 1a) and with visualization on MRI/more than 6 mm (group 1b). Methods: Immunohistochemically determined granulation pattern (group 2a densely granulated, 2b Crooke cell tumor) and proliferation levels based on Ki-67 with a threshold of 3% (group 3a high proliferation, 3b low proliferation), and biological behavior (group 4a indolent, 4b aggressive). Our study analyzed fresh tissue samples of CPitNETs from 34 patients with Cushing’s disease. Total RNA was extracted and sequenced using Cap Analysis Gene Expression (CAGE) on the Illumina HiSeq2500 platform, followed by bioinformatics analysis. Results: Differentially expressed genes were identified in group 1a tumors (EN1, EFS,) versus group1b (LCN1, BMP3). An inverse relationship was found between tumor volume and the expression of the RBM24 and ADGRD2 genes. A direct relationship was found between tumor volume and the expression of the PFN2 and CNTNAP5 genes. Identical gene signatures were found both in densely granulated and Crooke cell variants. Samples from group 3a, 4b and with bigger volume presented upregulated CT45A3, CT45A, CT45A1 genes. Samples from group 3b were enriched with PI3K/Akt pathway. Samples from group 4a showed expression of FXYD3, FGFR3 and FZD9 genes. Conclusions: Identified postgenomic markers in the studied subgroups of CPitNETs can potentially become biomarkers and allow to predict big volume and type of biological behavior at the early stages of diagnosis and treatment of patients with Cushing's disease.