Medical Technologies (Czechia)

Accurate monitoring of stress hormones is essential forunderstandingphysiological responses and optimizing human performance. Selectiveand precise detection of these biomarkers is particularly importantfor evaluating stress levels in high-demand environments. Herein,we report a multimodal sensor based on zinc oxide tetrapods (ZnO-Ts)for the detection of cortisol and testosterone. The platform integratesfluorescence and electrochemical transduction, enabling a complementaryand highly sensitive analysis. ZnO-Ts exhibited fluorescence quenchingupon interaction with steroid hormones with detection limits of 0.16μg/dL for cortisol and 0.25 μg/dL for testosterone. Stern–Volmerand time-resolved photoluminescence analyses revealed analyte-dependentquenching behavior, with cortisol exhibiting a dynamic quenching componentand testosterone showing static quenching. A solid-state electrochemicalsensor was further fabricated by embedding ZnO-Ts in a poly­(vinylalcohol) matrix and depositing them onto printed silver electrodes,which demonstrated concentration-dependent electrochemical responseswith higher sensitivity toward cortisol. By a combination of opticaland electrochemical readouts, this dual-mode sensing strategy providesrobust, rapid, and scalable hormone monitoring. The platform’sbiocompatibility and duality in identifying two steroidal hormonesselectively underscore its potential for wearable diagnostics andpoint-of-care stress monitoring technologies.

The oncolytic adenovirus Ad5-delta-24-RGD (Ad5Δ24RGD) exhibits suboptimal therapeutic efficacy in high-grade glioma. Here, we show that combining two specific potency-enhancing mutations in E3-19K and i-leader (designated 19KSS-iLQ125Ter) markedly increases the cytolytic efficiency and the size of vital dye-stained plaques. Unexpectedly, these mutations do not enhance actual spread efficiency, measured as the size of fluorescent plaques, in most tested cell lines. In contrast, the type of fiber modification (F5RGD4C, F5/3, or F5/35) strongly influences actual spread efficiency, with F5RGD4C generally conferring the greatest enhancement. In some cell lines, the fiber modification F5RGD10(2C) or verapamil treatment further improves actual spread efficiency. Nelfinavir inhibits spread and plaque formation of oncolytic adenoviruses with the 19KSS-iLQ125Ter modifications, irrespective of adenovirus death protein (ADP) expression. Expression of the reporter transgenes EGFP and Fluc or the human hyaluronidase PH20/SPAM1 from the E1B-55K region or downstream of the L3-23K or L5-Fiber region differentially affects the oncolytic potency of Ad5-delta-24-RGD-19KSS-iLQ125Ter. We identified an insertion site downstream of the L3-23K region that supports relatively high hPH20 activity while preserving the enhanced oncolytic potency of the virus. Combining 19KSS-iLQ125Ter with hPH20 expression may potentially improve therapeutic benefit in glioma.

Cardiac surgery patients often require allogeneic platelet transfusions. The blood service produces several types of donor platelet concentrates (РС), the methods of obtaining and additional processing of which directly affect the functional state of the cells and their clinical effectiveness. Pathogen reduction РC reduces the risk of transmission of infections (bacteria, viruses), but its effect on clinical effectiveness in cardiac surgery has not been sufficiently studied. Currently, there are no large randomized studies on this topic. Purpose of the study . To compare the effectiveness of the clinical use of different types of donor platelet concentrates in adult cardiac surgery patients. Material and methods . A retrospective randomized controlled trial was conducted. Adult cardiac surgery patients who received one or more РC units were eligible for inclusion. Exclusion criteria included patients with the presence of factors that could affect post-transfusion platelet growth: sepsis, febrile fever, РC transfusion resistance. The primary endpoint was an increase in the number of circulating peripheral blood platelets at 1 and 24 hours. The secondary endpoint was the need for repeat transfusions. The t test was used to compare continuous variables. Results . The study included 162 patients who were divided into groups that received either pathogen-reduced (group 1) or leukoreduced (group 2). There were no statistically significant differences between the groups. Discussion . In both groups, the number of platelets after 24 hours exceeded the initial level. However, the absolute increase in platelet count after 1 and 24 hours in group 1 was lower by 17 and 15%, respectively. In order to take into account the individual characteristics of patients, the method of adjusted platelet growth was used. After 1 hour and 24 hours indicated the effectiveness of replacement therapy, but in the first group it was lower than in the second. Lower platelet growth rates in the first group were not accompanied by a shortening of the interval between transfusions. Conclusions. Pathogen-reduced РC transfusions are associated with lower APT and SPT at 1 and 24 hours. However, a decrease in these indicators does not lead to a decrease in the interval between transfusions and an increase in the frequency of transfusions. The results obtained above may indicate equal clinical effectiveness of both types of РC.

Pseudogenes are non-functional copies of protein-coding genes that arise through genomic duplication or retrotransposition. Processed pseudogenes (PPs) is the most abundant class of pseudogenes, which is generated via mRNA reverse transcription and subsequent cDNA integration. Presence of PPs complicates the analysis of short read sequencing data due to high similarity with parental gene and frequent absence from reference genome. Here we demonstrate that the presence of non-reference (absent from reference genome) PPs leads to the very distinctive artefact of germline variant calling - long insertions on exon-intron boundaries, which sequences could be mapped to other exons of the same gene. We showed that by detecting these artifacts it is possible to identify non-reference PPs existence based on the cohort summary statistics without analysing samplelevel data. We used identified signature of PPs presence to systematically mine the gnomAD database which currently contains over 70,000 whole-genome and 1 over 700,000 exome samples to describe novel non-reference PPs. Our approach uncovered 1498 non-reference PPs of which 1268 were novel and absent in the latest GENCODE release. This resource enhances the accuracy of variant interpretation and contributes to a deeper understanding of pseudogenes diversity across human populations.

The identification of psychophysiological mechanisms underlying internal pronunciation in healthy individuals may enhance the performance of modern brain–computer interfaces that rely on the recognition of mental commands. Studying the brain mechanisms of internal pronunciation in people with stuttering can contribute to understanding the causes of this speech disorder and to developing new or improving existing methods of stuttering correction. The purpose of the study was to compare the psychophysiological mechanisms of internal speech (internal pronunciation) and word perception in people with normal and speech defects (stuttering). Electroencephalographic (EEG) recordings were obtained from 35 participants (25 with normal speech and 10 with stuttering) using a 19-channel system while they performed a task involving listening to and internally pronouncing four words in Russian (“sakhar”, “shashlyk”, “raketa”, and “r’aketa”). The analysis was carried out using a novel tool – the virtual implanted electrode technique (Patent RU 2,785,268 C1 by A.V. Vartanov), which enables the reconstruction of neural activity across several brain regions, including six areas of interest: Broca’s area, Wernicke’s area and their right-hemisphere homologues, as well as the cerebellum bilaterally. The activity data obtained were compared in control (normal) groups and people with stuttering. The comparison revealed significant differences in reconstructed electrical activity with an emphasis on hyperactivation in stutterers of the Broca’s area, the symmetrical zone on the right, and the cerebellar cortex bilaterally, compared with the control group, when listening and pronouncing words internally. In this way the process of perception and internal pronunciation of words in a group of normals and stutterers is different. The significant differences observed in Broca’s area likely reflect its primary role in speech production, whereas differences in the cerebellar cortex may be related to its supportive function in the motor coordination of speech. The absence of differences in the Wernicke’s zone can be considered as a consequence of the lack of auditory feedback during internal pronunciation.

Introduction. Early detection of patients with a high risk of death in sepsis makes it possible to optimize intensive care. The objective of the study was to perform a comparative assessment of the prognostic values of various laboratory indicators and their combinations in predicting mortality in patients with sepsis. Materials and methods. We analyzed the medical records of 280 patients diagnosed with sepsis. Levels of more than 50 laboratory parameters were assessed upon admission in groups of patients who survived and those who died during treatment. The discriminatory abilities of the identified laboratory predictors were assessed using ROC analysis. The Kaplan–Meier method was used to assess survival. Results. Among the studied patients, the mortality rate was 50%. Differences between the groups of surviving and deceased patients were identified according to levels of urea, total protein, albumin, lactate, pH, platelet and neutrophil count. The lowest level α (p < 0.001) was found for the ratio of urea to creatinine and lactate to albumin. Based on this, an integral biomarker consisting of the product of these two parameters was calculated and analyzed. Its values also differed between groups and amounted to 3.9 (2.9–6.8) and 9.8 (4.8–18.8) in the group of surviving and deceased patients, respectively (p < 0.001). When performing ROC analysis, the optimal cut-off point for our predictor, separating survivors from deceased patients, was 6.6 with an AUC of 0.758 (95% CI 0.693–0.813), with a sensitivity of 67% and specificity of 75%, exceeding similar indicators for the APACHE II scale. We also found that patients with low levels of our proposed predictor had a threefold increase in median survival compared to the subgroup of patients with high levels (23 days and 8 days, respectively; p < 0.001). Conclusion. An integrated indicator based on plasma levels of urea, creatinine, lactate, and albumin is a simple and accessible biomarker for early prediction of mortality in patients with sepsis.

A new dimeric dipeptide mimetic with antidepressant-like activity has been developed at the FSBSI "FRC of Original and Promising Biomedical and Pharmaceutical Technologies". A specific, sensitive, fast and reproducible method for the determination of bis -( N -monosuccinyl- L -asparaginyl- L -asparagine hexamethylenediamide (GTS-301) in rat blood plasma using high-performance liquid chromatography tandem mass spectrometric detector has been developed and validated. The lower limit of quantification in blood plasma was 50 ng/ml. All calibration curves were described by a quadratic equation (correlation coefficient averaged 0.996, n = 3) in the concentration range of 50÷10000 ng/ml. The accuracy values for the quality control samples varied throughout the day from an average of 93.2 to 113.3 %. The precision ranged from 1.1 to 11.4 %. During the validation period, the accuracy values averaged 101.1–108.8 % and accuracy — 5.4–7.3 %. For the calibration standards, the values of the validation parameters were also within acceptable values (accuracy — 92.2–106.8 %, accuracy — 0.1–11.5 %). The results of the dilution study of experimental blood plasma samples with intact biomaterial in 10and 100-fold lie within the acceptable values of 85–115 %. It has been established that GTS-301 is stable in the biological material at ambient temperature for short-term storage (3 hours), exposure to a thermostatically controlled autosampler (5 °C) during an analytical experiment (up to 20 hours), and prolonged storage at -50 °C (for 4 weeks), and also if subjected to several freezing/defrosting cycles (3 cycles). Thus, the above-mentioned characteristics of the developed technique make it suitable for use in preclinical pharmacokinetic studies. The pharmacokinetics of GTS-301 in rat blood plasma after a single intraperitoneal injection at a dose of 150 mg/kg was described.