Background. Malaria is a global public health problem. Traditional blood microscopy, the gold standard of diagnosis, is associated with high labor burden, personnel shortages in endemic regions, and subjective assessment. Automation of not only detection but also determination of the pathogen’s developmental stage is critically important for prescribing stage-specific therapy.Aim. To develop and evaluate the effectiveness of a deep computer vision model for classifying four key stages of the erythrocytic cycle of Plasmodium falciparum based on microphotographs.Materials and methods. A convolutional neural network with the ResNet-18 architecture was used as the basis. Training was conducted using transfer learning on the PfMorphoBank database, assembled by the authors. The images were divided into four target classes: young trophozoites, mature trophozoites, schizonts, and gametocytes. Classification quality was assessed on validation and held-out test sets using accuracy, macro-precision, macro-recall, and macro-F1.Results. On the held-out test set, the fine-tuned ResNet-18 achieved 80.83% accuracy; macro-precision was 0.812, macro-recall was 0.808, and macro-F1 was 0.806. Error analysis showed better recognition of mature trophozoites and the greatest difficulty with gametocytes and morphologically similar late-stage forms.Conclusion. The use of deep neural networks demonstrates high potential in solving the problem of differential diagnosis of malaria parasite stages. The model showed acceptable quality metrics, proving the fundamental feasibility of creating automated prompting systems.
Objective assessment of postoperative eye refraction in patients with implanted intraocular lenses (IOLs) featuring addition zones or a modified wavefront profile is challenging using conventional methods.Aim. To analyze the diagnostic value of the method of objective refractive topographic mapping of the eye in patients with IOLs of complex optical design.Material and Methods. A total of 198 patients (mean age 66.4 ± 7.8 years) with various types of IOLs – monofocal, multifocal, and extended depth of focus lenses were initially screened. Inclusion criteria: the absence of pathology of the cornea, vitreous, retina, or optic nerve; a postoperative follow-up period of six months or more. Exclusion criteria: uncorrected distance visual acuity of 0.7 or worse, opacity or folds of the posterior lens capsule. After applying inclusion and exclusion criteria and performing random eye selection (right or left) for eligible participants, 192 eyes were included in the final analysis. One eye per subject was chosen at random to avoid inter-eye correlation. All patients underwent a comprehensive ophthalmological examination, including objective assessment, visometry, tonometry, depth of focus (DoF) calculation based on the defocus curve (the diopter range within which visual acuity remains no lower than 0.5), wavefront aberrometry (Peramis, SCHWIND eye-tech-solutions GmbH, Germany) with refractive topographic map (RTM) construction and calculation of the refractive inhomogeneity index.Results. The construction of RTMs revealed the distribution of refractive indicators across the entire scanning area and the presence of distinct addition zones in eyes with multifocal IOLs and extended depth of focus IOLs. Significant differences were found in the refractive inhomogeneity index values between groups (H(7) = 46.24, p = 0.0001) and varying levels of its consistency with DoF.Conclusion. The method of pyramidal sensor-based aberrometry with refractive topographic mapping represents a highly informative approach for the objective assessment of the eye’s optical system in patients with complex-design IOLs and could serve as a foundation for the development of personalized cataract surgery.
Background. For the development of HER2-targeted CAR NK cell effectors, it is important to select an NK cells subset with high cytotoxic potential and sensitivity to genetic modification. In the context of solid tumor therapy, licensed KIR+ NK cells expressing the activating receptor NKG2C are of particular interest.Aim. To obtain HER2-CAR-NK cells based on NK cell subsets with different expression of KIR and NKG2C receptors for the elimination of HER2+ solid tumors.Materials and methods. NK cells from an HCMV-seropositive donor were isolated by negative magnetic separation from peripheral blood mononuclear cells isolated by density gradient centrifugation. NK cell subsets with different expression of KIR and NKG2C were obtained by cell sorting and cultured with stimulation by IL-2 and feeder cells. NK cells were modified with a gene carrying the HER2-CAR construct via retroviral transduction. The cytotoxic activity of the cultures against SKOV-3 spheroids was detected by the level of degranulating CD107a+ NK cells and the proportion of nonviable SytoxBlue+ tumor cells in the spheroid.Results. HER2-CAR-NK effectors were obtained based on subsets with different KIR and NKG2C expression. It was shown that NKG2C-expressing NK cells, regardless of KIR expression, have greater sensitivity to retroviral transduction. Upon co-incubation of the obtained cultures of NK cell subsets with HER2+ SKOV-3 tumor cell spheroids, in the fraction of transduced cells bearing the HER2-CAR receptor, the proportion of degranulated CD107a+ cells was more than 4-fold higher than the level of degranulation in the fraction of unmodified NK cells. Also, in SKOV-3 spheroids after co-incubation with KIR+ NKG2C+ transduced NK effectors, a lower proportion of viable cells was observed compared to spheroids that were incubated with KIR+ NKG2C– cultures.Conclusion. NKG2C+ NK cells exhibited greater sensitivity to transduction. Cultures of KIR+NKG2C+ HER2-CAR-NK cells eliminated HER2+ target cells more efficiently in a 3D model. These properties make NK cells coexpressing KIR and NKG2C a promising basis for the CAR-NK effectors development.
Introduction. Obesity in adolescents is a global medical and social problem associated with a progressive increase in incidence. The role of structural changes in the colonic microbial community in the pathogenesis of exogenous-constitutional obesity in adolescents remains insufficiently studied, and data on characteristic taxonomic alterations are contradictory.Aim. To study the characteristics of the qualitative and quantitative composition of the colonic microbiota in adolescents with obesity and normal body weight.Materials and methods. A cross-sectional comparative study was conducted. The main group consisted of 20 adolescents aged 12–17 years with obesity (BMI SDS ≥ +2.0), and the control group consisted of 20 adolescents with normal body weight. The taxonomic composition of the microbiota was examined by real-time PCR using the “Colonoflor-16” test system. Statistical analysis was performed using the Mann – Whitney and Fisher tests.Results. Compared to the control group, changes in the taxonomic composition of the microbiota in adolescents with obesity were characterized by a significant decrease in total bacterial mass and the abundance of key symbionts, including Bifidobacterium spp. A decrease in the level of Lactobacillus spp. was observed in both study groups without statistically significant intergroup differences. A deficiency of butyrate producers and key participants in fiber fermentation was identified, including Faecalibacterium prausnitzii, Eubacterium rectale, and Akkermansia muciniphila, as well as a trend toward a decrease in Roseburia inulinivorans, Ruminococcus spp., Prevotella spp., and Blautia spp. Concurrently, an increase was recorded in the abundance of Bacteroides spp., opportunistic pathogens (Enterobacter spp., Escherichia coli, Streptococcus spp., Enterococcus spp.), and the Bacteroides spp. / Faecalibacterium prausnitzii ratio. In the main group of adolescents, Clostridium difficile, Clostridium perfringens, Staphylococcus aureus, Acinetobacter spp., and fungi of the genus Candida were detected.Conclusion. Adolescents with obesity exhibit a specific pro-inflammatory profile of the colonic microbiota, characterized by depletion of symbionts, deficiency of butyrate producers, and overgrowth of opportunistic taxa. The obtained data identify potential targets for personalized microbiome correction in the comprehensive therapy of obesity.
Introduction. Serine proteases are the most numerous catalytic type of proteolytic enzymes and perform a variety of functions in the body. They participate in digestion, act as blood clotting factors, are components of complement and the kallikrein-kinin system, among others. Trypsin is the most studied serine protease. It is known that in certain pathological processes and conditions, such as pancreatitis and pancreatic necrosis, trypsin enters the systemic circulation. This can lead to the development of thrombosis and thromboembolism, but the mechanisms underlying these disorders remain poorly understood. One possible mechanism may be trypsin-mediated platelet activation followed by aggregation.Aim. To investigate the ability of trypsin to induce platelet aggregation in vitro. Methods. Experiments were conducted on platelet-rich plasma obtained from 22 relatively healthy volunteers. Platelet aggregation by trypsin at final concentrations of 0.5, 1, 2.5, 5, and 10 μM was studied using optical aggregometry. Trypsin-induced platelet aggregation was then recorded in prothrombin-deficient plasma.Results. Trypsin is capable of inducing platelet aggregation in vitro. The minimal effect was observed with the addition of 2.5 μM. Addition of 5 and 10 μM trypsin to platelet plasma induced pronounced and irreversible platelet aggregation. Trypsin-induced aggregation occurs in both heparinized plasma and plasma deficient in prothrombin complex factors.Conclusion. The study revealed proaggregatory effects of trypsin that are thrombin-independent and can occur without its participation.
