Learning and memory

Pulmonary tuberculosis (TB) is a socially significant disease and a global challenge faced by public health. The NF-kB signaling pathway is involved in differential expression of the genes involved in immune responses and regulation of inflammation in response to infection. The study aimed to assess associations of the NFKB1 allelic variants with TB based on the panel of SNPs (rs4648050, rs4648051, rs4648055, rs4648058, rs4648068, rs1609993) located within the NF-kB1 р105→р50 processing region. Total DNA was extracted from blood samples (phenol-chloroform extraction) of patients with TB (n = 93) and the population control group (n = 96) consisting of residents of the Kemerovo Region. Genotyping was performed by real-time PCR, and the results were processed using the resources of the Statictica, SNPStats, Arlequin software packages. Ethnic features (p < 0.05) of the Russian population of Siberia (population control group) were demonstrated based on the rs4648050 and rs4648051 allele frequencies. Differences (p < 0.05) of the genetic profile of the sample of patients with pulmonary tuberculosis throughout the entire SNP complex, except for rs1609993, were noted. We showed differences (p < 0.05) in the rs4648068 allelic frequencies between the population control sample and patients with pulmonary tuberculosis. The association with susceptibility to pulmonary tuberculosis was determined for genotypes АА*rs4648055 (OR = 2.51; p = 0.05) and GG*rs4648068 (OR = 2.16; p = 0.03). The findings are indirect evidence of modifying effects of the SNP located within the processing zone in the gene NFKB1 and its possible contribution to the NF-kB1 р105/р50 protein balance and immune response to mycobacterial infection.

The COVID-19 pandemic necessitated making timely managerial decisions when providing medical care to patients with tuberculosis (TB). The study aimed to develop a system for monitoring of TB combined with COVID-19 and estimate the prevalence of COVID-19 among TB patients, along with the efficacy of the measures applied. A registry of TB/COVID-19 patients was developed based on the Barclay-SV Medical Database Management System. It was used to perform comparative analysis of the information about 1837 patients with active TB forms and confirmed COVID-19 for two periods of the pandemic, 2020–2021 and 2022–2023, and against the data on all new TB cases and TB relapses registered in Moscow in 2020–2023: 7812 and 1243 individuals respectively, from the TB surveillance registries, excluding those identified posthumously. The socio-demographic structure of patients with TB/COVID-19 co-infection identified in 2020–2023 did not change and corresponded to that of TB patients. In the second period analyzed, mild COVID-19 cases were registered more often (60.9% vs. 41.6%; p < 0.01), the share of moderate COVID-19 cases decreased from 48.2% to 20.6% (p < 0.01), and the share of severe cases decreased from 6.4% to 4.9% (p = 0.19). In 2022–2023, the share of individuals with COVID lung damage decreased from 45.1% to 17.6%, while the number of cases of COVID upper respiratory tract lesion increased from 47.1% to 64.5% (p < 0.05). The fact of having HIV infection, CAD and hypertension, kidney and genitourinary diseases increased the chance of developing COVID-19 by TB patients 1.5–2-fold, and disseminated pulmonary tuberculosis, caseous pneumonia, lung tissue destruction and bacterial excretion increased it 1.4–1.6-fold. The registry made it possible to control routing of TB/COVID-19 patients, as well as treatment outcomes: the total share of individuals cured reached 90.1%.

The choice of the sterilization method for ceramic implants is critically important, as it can affect the chemical and physico-mechanical properties of the material and its biocompatibility. Higher cytotoxicity, which is a possible side effect of sterilization, hinders osseointegration. This study aimed to determine the cytotoxicity of porous ceramic samples after sterilization using the most common methods. Samples of hydroxyapatite (HA), tricalcium phosphate (TCP), and aluminum oxide (AO) were prepared by stereolithography, and bone allograph samples were made using the DLP method. The annealing lasted for 4 hours, with a peak temperature of 800 °C and the temperature increment of 3 °C per minute; the sintering temperature was up to 1200 °C. We used the following sterilization methods: autoclaving at 1 atmosphere, 120 °C, for 45 minutes; radiation sterilization, 25 seconds with an absorbed dose of 25 kGy; plasma peroxide sterilization, 42 minutes; dry heat sterilization at 180 °C, for 60 minutes. Cytotoxicity was determined with the help of an MTT assay (24-hour exposure in a CO2 incubator). The results of the study: for HA, high porosity means growth of values in transition from autoclaving (0.1115) to plasma peroxide sterilization (0.2023). Medium and low porosity show similar results, with peaks in dry-heat sterilization (0.4954 and 0.4505). As for for AO, it exhibited high viability when subjected to this method. The TCP samples have shown stable results, but their low-porosity variation had the values growing after autoclaving (0.078 to 0.182, dry-heat sterilization). The study forms the basis for optimizing the ceramic implants manufacturing technology and sterilization methods to ensure their high biocompatibility.

The widespread use of antibiotics in medicine and agriculture has significantly accelerated the emergence rate of bacterial infections showing multiple antibiotic resistance. Since resistance to conventional antibiotics is developed rather quickly, designing alternative antimicrobial drugs with other mechanisms underlying their effects on bacteria is a promising. The enzymes possessing bactericidal activity may be one option for such antibacterial agents. The study aimed to produce the combination recombinant protein-based products active against bacteria and their biofilms. Soluble forms of five recombinant proteins were produced using the genetic engineering approaches. Two of these have a bacteriolytic effect (endolysins LysK and PM9 from the Staphylococcus aureus bacteriophages), the other are capable of disrupting extracellular DNA matrix in biofilms (two nonspecific nucleases NucA, as well as the DNA-specific deoxyribonuclease I). It has been shown that natural endolysin PM9 with the truncated catalytic domain shows 4 times lower bacteriolytic efficacy compared to the full-size LysK version. Comparative analysis revealed 1.5–2 timed higher efficacy of nonspecific nucleases in terms of bacterial biofilm disruption compared to the DNA-specific deoxyribonuclease I. It has been shown that simultaneous use of endolysins and nucleases has a synergistic antibacterial effect and disrupts biofilms of the pathogenic bacterium Staphylococcus aureus. The findings show the prospects of developing the recombinant protein-based antibacterial drugs.

Diabetes mellitus (DM) is one of the major factors contributing to the development and aggravation of chronic kidney disease (CKD). The accurate and convenient markers for early detection, estimation of progression, and adequate control of CKD therapy in individuals with DM are limited to glomerular filtration rate (GFR) and albuminuria. Given the role of chronic inflammation in the pathogenesis of DM and CKD, the study aimed to assess indicators of inflammation and the correlation of those with GFR in patients with type 1 DM (T1D) and early stage CKD. The study involved healthy individuals (n = 14), patients with T1D showing no signs of CKD (n = 30), as well as patients with T1D and stage 1 CKD (n = 60), stage 2 CKD (n = 38), and stage 3 CKD (n = 31). GFR was calculated using the formula СКD-ЕРI (eGFR); serum levels of IL1β and TNFα, C-reactive protein (CRP), and ceruloplasmin (CP) were determined by enzyme immunoassay; the neutrophil-to-lymphocyte index and the leukocyte intoxication index (LII) were calculated. It has been found that serum concentrations of IL1β, TNFα, CRP, and CP are elevated; LII and the neutrophil-to-lymphocyte index are increased. The inflammation and acute phase response severity progresses and reaches its maximum in stage 3b CKD, when the serum concentration of IL1β is increased 2.4-fold (р = 0.042), TNFα concentration by 34% (р = 0.005), CRP concentration 33-fold (р < 0.000), CP concentration by 73% (р = 0.008), LII 8.4-fold (р < 0.000), neutrophil-to-lymphocyte index 5-fold (р = 0.013). The integral kidney function indicator, eGFR, decreases with increasing serum levels of the above indicators. Thus, IL1β, TNFα, CRP, CP, LII, and the neutrophil-to-lymphocyte index can be considered as affordable and informative indicators for estimation of inflammation, the levels of which increase with progression of early stage CKD in patients with T1D.

Staphylococcus aureus is the causative agent of a wide range of infections, including severe systemic diseases, which is often multidrug resistant. Given the growing overall antibiotic resistance, a promising approach to treating staphylococcal infections is administration of bacteriophages, especially in combination with antibiotics. This study aimed to evaluate the synergistic effect of linezolid and bacteriophage vB_SauM-515A1 in combating a systemic infection in BALB/c mice. Using 36 animals, we established the optimal way of administration and the infecting dose of the microorganism (5 × 108 CFU/mouse intravenously), and identified the threshold concentrations of antimicrobial agents for monotherapy. The evaluation was based on the revealed contamination of internal organs (kidneys, spleen) and blood. To learn the etiotropic effect of linezolid (10 mg/kg animal weight) combined with the phage (2 × 107 PFU/mouse), we worked with a control group and a test group, 12 mice in each; 2, 8, 18, and 24 hours after infection, the former received the drug only, the latter — the investigated combination. Combined therapy had a more pronounced effect, decreasing the bacterial load in the kidneys by two to three orders of magnitude compared with monotherapy on the first day of treatment. Thus, the combined use of linezolid and bacteriophages is promising for the treatment of infections caused by S. aureus, and may increase the effectiveness of treatment and reduce the risk of side effects of high-dose antibiotics.

Calculation of toric intraocular lenses (tIOLs) in patients after penetrating keratoplasty (PK) is challenging. The study aimed to perform comparative retrospective analysis of various methods for calculation of tIOL during phacoemulsification in patients after PK. We analyzed case reports of 36 eyes (36 patients) after PK, which underwent phacoemulsification with tIOL implantation. All tIOLs were recalculated using four different methods. In group 1, tIOL calculation was performed using keratometry data of the anterior surface of the corneal graft measured using a corneal topographer, and the posterior surface of the corneal graft measured using optical coherence tomography of the cornea or the Scheimpflug keratotopographer. In group 2, keratometry of both corneal graft surfaces was measured using the Scheimpflug keratotopographer, in group 3 — using OCT of the cornea, in group 4 — using the keratotopographer. The online Barrett True — K Toric Calculator was used to calculate tIOLs in groups 1–3, and The Kane Formula was used in group 4. There were significant differences in the values of the spherical and cylindrical components of refraction between the studied groups (p < 0.05). The highest predictability of tIOL calculation was reported for group 1: the ensured postoperative refraction for the spherical component was within ±0.5 D in 58% of eyes, within ±1.0 D in 67% of eyes; postoperative refraction for the cylindrical component was within –0.5 D in 56% of eyes, within ‒1.0 D in 89% of eyes. Thus, the highest predictability of tIOL calculation is observed in patients of group 1.

Staphylococcus aureus causes a broad range of infections and is often characterized by multidrug resistance (MDR). Treatment of staphylococcal infections is further complicated by the ability of bacterium to form biofilms protecting it against antimicrobial agents and the immune system. The use of bacteriophages is one of the promising strategies for combating the bacteria showing MDR and biofilm formation activity. The study aimed to assess the effects of the lytic phages vB_SauM515A1 (genus Kayvirus, family Herelleviridae) and vB_SauP-436A (genus Rosenblumvirus, family Rountreeviridae) on biofilms of the S. aureus clinical strains. The study involved 20 strains of eight sequence types, among which 45% (9/20) belonged to MRSA, and 35% (7/20) showed MDR. All the strains demonstrated the ability to form biofilms, and 65% (13/20) were strong biofilm producers. Genes of the icaADBC operon responsible for synthesis of polysaccharide intercellular adhesin were found in genomes of all samples. The exposure of planktonic bacterial cells to bacteriophages showed that 70% (14/20) of strains were sensitive to phage vB_SauM-515A1 and 50% (10/20) were sensitive to phage vB_SauP-436A. Furthermore, the 24-h treatment of biofilms of sensitive strains with phage vB_SauM-515A1 led to the biofilm biomass increase in 64.3% (9/14) of cases, while phage vB_SauP-436A, on the contrary, significantly reduced the quantity of biofilm in 40% (4/10) of strains. The results obtained highlight the ambiguity of interaction between bacteriophages and S. aureus biofilms and suggest the need for further research aimed at optimizing phage therapy targeting the biofilm-forming strains.

Today, preoperative hormone therapy is a standard procedure in the context of treatment of ESR+/HER2-negative early-stage breast cancer. Transcription profiles of genes helps make assessment of effectiveness of this therapy more accurate. This study aimed to investigate the changes in gene expression caused by the preoperative aromatase inhibitor response test in postmenopausal women with ESR+/HER2-negative breast cancer. The participants were 100 breast cancer patients treated at the Department of Breast Pathology of Kulakov National Medical Research Center for Obstetrics, Gynecology and Perinatology. We did a pathomorphological study of FFPE blocks (trephine biopsied before the hormone response test was prescribed) and intraoperative samples, and immunohistochemical (Ki67, ER, PR, HER2/neu) and molecular genetic studies of 45 target genes (quantitative RT-PCR). Aromatase inhibitors in the preoperative hormone response test caused significant changes in the mRNA expression of 37 genes in breast tumors: for 35 of them (ESR1, PGR, AR, ERBB2, FGFR4, MKI67, MYBL2, CCNB1, AURKA, BIRC5, CCND1, CCNE1, CDKN2A, KIF14, PPP2R2A, PTTG1, TMEM45B, TPX2, ANLN, MMP11, CTSL2, EMSY, PAK1, BCL2, BAG1, PTEN, TYMS, EXO1, UBE2T, NAT1, SCGB2A2, GATA3, FOXA1, ZNF703, CD274/PD-L1) the level was decreased, and for 2 genes it increased (SFRP1, KRT5). The results of this study can be used in the development of a hormone sensitivity test and personification of adjuvant systemic treatment for breast cancer patients.

As M. tuberculosis strains develop resistance to fluoroquinolones, pools of M. tuberculosis sensitive to drugs of this group and pools of M. tuberculosis with different resistance determinants can simultaneously coexist in the host organism. The goal of this research was to run an in vitro investigation of growth characteristics of M. tuberculosis strains which have different genetic determinants of resistance to fluoroquinolones, in the setting of competition for nutrients. The research used five clinical strains of multidrug-resistant M. tuberculosis differing in gyrA structure. Strains were cultured in pairs and individually under optimal conditions (Middlebrook 7H9 medium) and under conditions of multistress (50% Middlebrook 7H9 medium, 2 mM KNO2, 0.02% H2O2). The experiment took 21 days. The number of cells of each co-cultured strain was estimated from calibration curves. These curves showed the dependence of the threshold cycle of the polymerase chain reaction — respective to the channel targeted by the mutation — on the concentration of M. tuberculosis cells. The competitive fitness value and specific growth rate were calculated from the number of cells of each strain when co-cultured. M. tuberculosis strains with mutations in gyrA were found to be inferior in growth rate to the wild-type gyrA strain, which was particularly pronounced under multistress conditions. The strain with the most common gyrA_D94G mutation had the lowest growth rate of all strains examined. It has been hypothesised that the slow growth of M. tuberculosis with this mutation may lead to tolerance to anti-tuberculosis drugs, and as a result, the strain gains an advantage under chemotherapy conditions compared to other gyrA mutant variants.

Today, implantation of an intraocular lens (IOL) into the capsular bag is the standard approach to surgical treatment of cataracts and aphakia of various origins. However, there are several reasons and conditions that disallow this operation or increase the risk of instability of the implanted lens, such reasons and conditions including weakness of the lens ligaments; degradation of Zinn's zonule, including dislocation of the IOL‒capsular bag complex post-surgery; damage to or removal of capsular bag during surgery; lack of capsular bag or its destruction during implantation in aphakia cases. To date, problems associated with fixation and centralization of IOL in non-standard cases involving weak or inexistent capsular support remain unresolved. This study aimed to develop techniques allowing to stitch IOL to the iris without compromising its functions in various situations when it is unfeasible or impossible to fix and center lens in the capsular bag. The patients (n = 12; 12 eyes), depending on the clinical situation, were divided into groups: group 1 — dislocations of the IOL–capsular bag complex (6 eyes); group 2 — complete lack of capsular support (3 eyes); group 3 — weakness of capsular support (3 eyes). A special stitching technique was developed for each of these situations. The results of the treatment were good from clinical and functional perspectives: the IOL was fixed securely and centered properly, and the iris's performance and cosmetic aspects were not compromised.

Multiple sclerosis (MS) is a chronic disorder of the central nervous system affecting primarily young women. Neurogenic lower urinary tract dysfunction (NLUTD) represents one of the disease manifestations creating the risk of infectious complications and kidney disease. Today, there is insufficient data on the urinary microflora composition obtained by advanced high-tech diagnosis methods. The study aimed to perform clinical assessment of NLUTD associated with MS and its impact on the quality of life (QOL), as well as to clarify the data on the urinary microflora composition. A total of 33 women with MS aged 36 [39.5; 30.5] years were assessed using the customized questionnaires for estimation of the NLUTD prevalence and severity, as well as for QOL evaluation. Qualitative determination and quantification of urinary opportunistic microflora (OM) were performed using the real-time polymerase chain reaction. A total of 19 (57.6%) women with MS had symptoms of NLUTD: symptoms of the storage (15 individuals, 45.5%) and emptying (16 individuals, 48.5%) phases. In almost half of women with MS, the complaints included abnormalities of both bladder functioning phases (12 individuals, 36.4%); moderate abnormalities prevailed (12 individuals, 34.6%). Women with MS and NLUTD were more disabled based on the EDSS score (3.5 [5.0; 3.0] points; p < 0.001) and had longer disease duration (13 [20.0; 5.0] years; p < 0.001). The QOL index of women with NLUTD showed dissatisfaction with bladder function. The study revealed bacteriuria in patients with MS and NLUTD. The data on the urinary microflora composition are provided: OM members (bacteria of the ESKAPE group) have been found in 8 samples obtained from women with MS and NLUTD. Bacteriuria was asymptomatic.

Residents of the Techa Riverside villages were chronically exposed to the wide range of doses more than 60 years ago. Telomeric regions of metaphase chromosomes in the cultured peripheral blood T-lymphocytes were the subject of the research. The study aimed to assess the impact of chronic exposure on telomere loss in exposed women of the Southern Urals using a fluorescent staining method. Chromatid and chromosome telomere loss was determined in three dose subgroups: comparison group (0–0.01 Gy), group of exposed individuals with the dose of 0.2–0.9 Gy, and group of the exposed individuals with the dose of 1–4.6 Gy. In the sample of female residents of the Southern Urals chronically exposed in the range of absorbed doses to RBM of 0–4.6 Gy, it was shown that there were no differences in telomere loss between the comparison group and the group exposed to the dose exceeding 1 Gy (p > 0.33), while the group of individuals exposed to medium doses of 0.2–0.9 Gy was statistically significantly different (p < 0.05). Statistically significant differences between all groups were reported for chromosome telomere loss (p < 0.05). According to the data obtained, telomere loss was found in 99.85% of donor cells. The loss of telomere region on one of the chromatids occurred statistically significantly more often in all the groups. Thus, in the group exposed to the dose of 0.2–0.9 Gy, the average rate of chromatid telomere loss was higher, it was statistically significantly different from that of the other groups of females of the studied age.

Erectile dysfunction (ED), an unusual sexual condition in which the person fails to attain or sustain an erectile penis, severely impacts personal relationships, confidence, and efficiency. To date, low-intensity extracorporeal shock wave therapy (Li-ESWT) is an option to manage ED; however, it is associated with adverse events such as bruising, redness, and pain. Hence, in this study, we applied platelet-rich plasma (PRP), a blood-derived biomaterial containing cargo of growth factors, to enhance the therapeutic efficacy of Li-ESWT on ED. We assessed the synergistic effect of PRP+Li-ESWT, in which Li-ESWT was extracorporeally applied simultaneously with PRP. They were evaluated clinically at 22 ± 2, 50 ± 2 and 78 ± 2 days. Statistical analysis was performed using a non-parametric test, Friedman repeated measures as an alternative non-parametric test of ANOVA test. The international index of erectile function (IIEF-5) and erection hardness score (EHS) were recorded. IIEF-5 score in the pre-treated group was 8.36 ± 1.44. After 22 ± 2 days of synergistic PRP+Li-ESWT treatment, the score was 14.45 ± 2.12 (p < 0.028). This score further increased to 15.45 ± 1.93 (p < 0.008) and 16.18 ± 1.48 (p < 0.001) after 50 ± 2 days and 78 ± 2 days of treatment, respectively. The mean pre-treated EHS was 1.64 ± 0.20 (p < 0.002), which increased to 2.81 ± 0.26 (p < 0.002), 3.09 ± 0.25 (p < 0.0002) and 3.18 ± 0.12 (p < 0.000) on day 22 ± 2, 50 ± 2 and 78 ± 2 days, respectively. Conclusively, our study demonstrated potent synergistic therapy of PRP+Li-ESWT in ED treatment by improving IIEF-5 and EHS scores. However, extensive mechanism-based clinical studies are needed to reach a consensus.

Immunotherapy with oncolytic viruses (OVs) becomes a full-fledged neoadjuvant therapy method in the paradigm of evidence-based medicine for the growing number of cancers. The use of OVs for immunologically “cold” tumors causing minimal immune response and having the clearly immunosuppressive tumor microenvironment is especially relevant. Recombinant OVs carrying the sequences of proteins activating the immune system can be used to stimulate antitumor response. The study aimed to assess oncoselectivity and antitumor activity of the recombinant OV designed based on the LIVP vaccinia virus strain showing expression of human and murine interpheron alpha sequences (hIFNα and mIFNα, respectively). The in vitro experiments showed that the recombinant OVs designed showed oncoselectivity in relation to tumor cell lines of appropriate species. The ability to effectively infect human adenocarcinoma and glioblastoma cell lines was reported for LIVP-hIFNα. LIVP-mIFNα showed selectivity in relation to glioma Gl261 and melanoma B16 in vitro. The in vivo experiment involving the C57Bl/6 mice with subcutaneous melanoma В16 showed the ability of the intravenously administered LIVP-mIFNα to reduce the size of the subcutaneous tumor allograft and increase tumor infiltration with the CD8+ and NK cells. The recombinant virus designed can be a potential platform for the development of oncolytic virotherapy of human melanoma and glioblastoma.