The Influence of the Synthetic Organoselenium Preparation 974zh on the Immunogenic Activity of Yersinia pestis EV Vaccine Strain NIIEG.

In the present study, a stimulating effect of a new synthetic organoselenium compound 2,6-dipyridinium-9-selenabicyclo[3.3.1]nonandibromide (974zh) on the immunogenic activity of the vaccine strain Yersinia pestis EV NIIEG was revealed. After infection with the virulent plague strain, the survival rate of laboratory mice immunized with the vaccine strain grown on Hottinger's agar in the presence of 974zh (300 µg/ml) increased in comparison with control animals immunized with the Y. pestis EV NIIEG culture grown on agar without the studied compound. Plasmid screening of cultures grown on medium with and without 974zh showed that plasmid DNA of Y. pestis EV culture grown in the presence of 974zh had broader bands in the control grown without 974zh. This phenomenon can indicate activation of replication of plasmid DNA of Y. pestis EV under the influence of the experimental compound.

High-Affinity Single-Domain Antibodies for Analyzing Human Apo- and Holo-Transferrin.

A highly efficient technology for generating new monoclonal single-domain recombinant antibodies (nanobodies) was used to obtain a panel of nanobodies recognizing human apo- and/or holo-transferrin. This article is devoted to the primary analysis of the properties of two different variants of the new nanobodies obtained by us, as well as to the demonstration of the unique potential of their application for diagnostic studies. The simultaneous use of immunosorbents based on these nanobodies apparently makes it possible to detect changes in the relative abundance of apo- and holo-transferrin in human biological fluids. Such changes could potentially be indicative of an increased risk or degree of development of pathological processes, such as malignant neoplasms in humans.

The effects of slow breathing on postural muscles during standing perturbations in young adults.

Maintaining standing balance is vital to completing activities in daily living. Recent findings suggest an interaction between cardiovascular and postural control systems. Volitional slow breathing can modulate the cardiovascular response and affect postural control during quiet standing. However, the effects of slow breathing during threats to standing balance have not been studied. The study examined the effect of slow breathing on the latency and amplitude of postural muscle responses to perturbations of the base of support in healthy, young adults. Twenty-seven participants completed two balance perturbation tasks in standing on an instrumented split-belt treadmill while breathing spontaneously and breathing at 6 breaths per minute. Each perturbation task consisted of 25 posteriorly directed translations of the treadmill belts every 8-12 s. Muscle latency and muscle burst amplitude were measured using surface electromyography from the right limb for the quadriceps (QUADS), medial hamstring (MH), gastrocnemii (GASTROC), soleus (SOL), and tibialis anterior (TA) muscle groups, while a respiratory belt was used to record respiratory rate. Results indicated that during the slow breathing task both muscle latency (p = 0.022) and muscle burst amplitude (p = 0.011) decreased compared to spontaneous breathing. The EMG pre-perturbation activation was not significantly different in any muscle group between conditions (p > 0.167). The study found that reducing respiratory rate to approximately 6 breaths per minute affects the neuromuscular responses in the lower limb muscles to perturbations.

[Single-Domain Antibodies Used to Pretreat the Human Urinary Proteome in Cancer Biomarker Testing].

A number of single-domain antibodies (nanobodies) obtained previously to major marker blood proteins were tested as tools to preprocess urine samples from patients with bladder cancer. Nanobody-based tools demonstrated unique possibilities for noninvasive diagnostic studies along with other conventional methods, such as electrophoresis and, in prospect, mass spectrometric analysis. A testing of 22 samples from bladder cancer patients showed that the development of bladder cancer is accompanied by an increase in the urine contents of major blood proteins, including those known as potential bladder cancer biomarkers. New nanobody-based immunosorbents allow both specific enrichment and specific removal of particular antigenic proteins and subproteomes associated with them from a biological fluid. The isolation of immune complexes from the urine of a particular patient is of particular interest. An initial study of the complexes showed not only increased contents of IgA and IgG at advanced stages of the disease, but also many other components, which provide potential biomarkers of the pathological process in a particular patient. It is intended to use the approaches proposed in this work in a future larger-scale study of urine samples from patients with bladder cancer at different stages of the disease in order to identify new promising biomarkers of bladder cancer.

Evaluation of the potential pathogenicity of microorganisms associated with urinary calculi.

Due to the prevalence of postoperative complications in the treatment of urolithiasis, the study of the contamination of urinary calculi and the potential pathogenicity of isolated bacteria is of great importance in laboratory diagnostic practice. It has been shown that uropathogenic bacteria are found in the composition of urinary stones in 65±7.1% of cases, mainly representatives of the Enterobacteriaceae and Staphylococcaceae families. Bacteria of the generas Escherichia, Enterococcus, Staphylococcus were most frequently detected. The analysis of biofilm activity and antibiotic resistance in 50 uropathogenic strains was carried out. It was shown that all the studied strains were resistant to at least two tested drugs, and the average value of the multiple resistance index was 0.51. When cultured on nutrient agar with Congo red, it was shown that more than half of the tested strains have high biofilm activity and about 80% potential for biofilm formation. The greatest biofilm activity was observed in bacteria of the generas Escherichia, Klebsiella, Enterobacter, Staphylococcus.

Use of SEM/EDX methods for the analysis of ambient particulate matter adhering to the skin surface.

BACKGROUND: The skin is exposed to numerous particulate and gaseous air pollutants. The ones that need particular attention are the particles that adhere to the skin surface, which can later cause direct skin damage. This study aimed to characterize air pollution (AP) particles adhered to the human skin by using scanning electron microscopy (SEM) combined with X-ray dispersive energy spectrometry (EDX). METHODS: Tape stripping was performed from six healthy volunteers exposed to urban AP to collect stratum corneum samples from the cheeks and forehead. The samples were analysed using SEM equipped with EDX system with a silicon drift detector at an accelerating voltage of 20 keV. After the preliminary examination, the particles were located and counted using 1000× magnification. Each particle was analysed, increasing magnification up to 5000× for precise dimension measurement and elemental composition analysis. At least 100 fields or a surface of approximately 1 mm2 were examined. RESULTS: Particles adhered to the skin were identified in all samples, with a particle load ranging from 729 to 4525. The average area and perimeter of all particles identified were 302 ± 260 µm2 and 51 ± 23 µm subsequently, while the equivalent circular diameter was, on average, 14 ± 6 µm. The particles were classified into ten groups based on morphology and elemental composition. Chlorides were the most numerous particle group (21.9%), followed by carbonaceous organic particles (20.3%), silicates (18%), carbonates (16.4%), metal-rich particles (14%), and a minor number of bioaerosols, quartz-like, and fly ash particles. CONCLUSION: The SEM-EDX analysis provides evidence of the contamination of exposed skin to various airborne PM of natural or anthropogenic origin. This method may provide new insights into the link between exposure to AP and AP-induced skin damage.

Postural control in response to unilateral and bilateral external perturbations in older adults.

BACKGROUND: Balance is an important determinant of physical function and falls risk. The ability to withstand external perturbations is important when walking on icy or uneven surfaces, whether the perturbations are bilateral or unilateral. RESEARCH QUESTION: This study sought to determine the effect of unilateral and bilateral standing perturbations on leg muscle activity in healthy older adults. METHODS: Participants experienced unilateral and bilateral standing perturbations of the treadmill. Surface electromyography (EMG) from lower limb muscles was recorded unilaterally. EMG onset latency and root mean square (RMS) amplitude of the muscle bursts were calculated. RESULTS: Older adults demonstrated a combined ankle/hip strategy, along with pre-activation and co-contraction of muscles in response to unilateral and bilateral stance perturbations. As well, older adults demonstrated higher levels of EMG, but no difference in the latency of burst onset, in bilateral than unilateral perturbation types. SIGNIFICANCE: When the stance limb was perturbed in the bilateral condition, the older adults responded with a Gastrocs EMG burst nearly 100% of the maximum EMG. The high level of EMG used, especially in the Gastrocs, during the bilateral perturbations may reduce the safety factor for falls in older adults. Older adults responded to the different perturbation demands by modulating EMG amplitude as opposed to the onset timing of EMG.

[Comparative Analysis of MPTP Neurotoxicity in Mice with a Constitutive Knockout of the α-Synuclein Gene].

Aggregated forms of α-synuclein are core components of pathohistological inclusions known as Lewy bodies in substantia nigra (SN) neurons of patients with Parkinson's disease (PD). The role of α-synuclein in selective loss of SN dopaminergic neurons (DNs) in PD is studied in mice knocked out in the α-synuclein gene. The new mouse strain delta flox KO with a constitutive knockout of the α-synuclein gene models the end point of in vivo deletion of the α-synuclein gene in mice with a conditional knockout and has no foreign sequence in the modified genomic locus, thus differing from all other α-synuclein knockout mouse strains. The effect of the neurotoxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP), which is used to model PD, was compared between delta flox KO mice and mice of the well-known α-synuclein knockout strain AbKO. Subchronic MPTP administration, which models early PD, was found to reduce the dopamine content and to change the ratio of dopamine metabolites in the striatum to the same levels in delta flox KO, АbKO, and wild-type mice. Overt locomotor defects were not observed after MPTP treatment, but gait testing in a CatWalk XT (Noldus) system revealed identical gait deviations in mice of the two strains and control wild-type mice. Based on the findings, a similar mechanism of neurotoxic damage to DNs was assumed for delta flox KO and AbKO mice.

Evaluation of the Association of rs4680 COMT Polymorphism and Clinical Parameters of the Tumor in Colorectal Cancer (a Pilot Study).

We analyzed association of rs4680 polymorphism of catechol-O-methyltransferase (COMT) gene with clinical parameters of the tumor in patients with colorectal cancer (n=100). Based on the classification of the tumor according to the TNM system, groups were formed taking into account the size and spreading of the primary tumor (T1+T2 vs Т3+Т4) and the presence of regional (N0 vs N1) and distant metastases (M0 vs M1). An association of the AA genotype with an almost 7-fold increased capacity for invasive tumor growth was found (p<0.05; according to recessive (AA vs GG+GA) and codominant (AA vs GG) inheritance models). A positive relationship of minor allele A with increased malignancy of tumor cells was revealed at the trend level. No significant associations with either regional or distant metastasis were found.

Cognitive Enhancer Noopept Activates Transcription Factor HIF-1.

The effect of noopept (N-phenylacetyl-L-prolyl-glycine ethyl ester) on the DNA-binding activity of HIF-1 in SH-SH5Y cells and the mechanisms of stabilization of this transcription factor were studied in vitro. Noopept was shown to increase both the basal DNA-binding activity of HIF-1 and the activity induced by various hypoxia mimetics. The mechanism of stabilization of the oxygen-sensitive HIF1α subunit by noopept involves the inhibition of HIF-1 prolyl hydroxylase, which is indirectly indicated by the data obtained using the ODD-Luc reporter, and the positive effect on the level of the HIF1α protein. It was revealed that the effect of noopept is accompanied by changes in gene expression, which belong to different metabolic pathways and are controlled by the transcription factor HIF-1.

Effect of Neuropeptide Cyclo-L-Prolylglycine on Cell Proliferative Activity.

Endogenous neuropeptide cyclo-L-prolylglycine possesses mnemotropic and neuroprotective properties, which can result from its positive effect on the level of brain-derived neurotrophic factor and modulation of activity of insulin-like growth factor-1 and AMPA receptors. For detection of possible mitogenic action of cyclo-L-prolylglycine, we analyzed its effect on proliferative activity of HEK293 and SH-SY5Y cells assessed by expression of Ki-67 proliferation marker, cell cycle examination, and incorporation of modified nucleotide analog EdU into DNA. Cyclo-L-prolylglycine did not affect the level of Ki-67 in examined cell lines and distribution of the cells over G1 and G2 phases of the cell cycle, although it insignificantly reduced the percentage of S phase cells, which attested to the absence of intrinsic mitogenic activity of the peptide. At the same time, cyclo-L-prolylglycine reduced the number of the early apoptotic cells, which can be a mechanisms of its protective action.

Immunotropic Properties of an Experimental Synthetic Selenium-Organic Compound.

We studied immunotropic properties of synthetic selenium-organic preparation 2,6-dipyridinium-9-selenabicyclo[3.3.1]nonyl dibromide (974zh). The experimental preparation reduced the cAMP/cGMP ratio, which indicated an increase in proliferative activity of cells of immunocompetent organs (thymus and spleen) in experimental animals. It was shown that 974zh intensified the immune response to Yersinia pestis EV thereby increasing the resistance to the plague agent.

Feedback-Induced Quantum Phase Transitions Using Weak Measurements.

We show that applying feedback and weak measurements to a quantum system induces phase transitions beyond the dissipative ones. Feedback enables controlling essentially quantum properties of the transition, i.e., its critical exponent, as it is driven by the fundamental quantum fluctuations due to measurement. Feedback provides the non-Markovianity and nonlinearity to the hybrid quantum-classical system, and enables simulating effects similar to spin-bath problems and Floquet time crystals with tunable long-range (long-memory) interactions.

Regional modulation of the ankle plantarflexor muscles associated with standing external perturbations across different directions.

Maintenance of upright standing posture has often been explained using the inverted pendulum model. This model considers the ankle plantarflexors to act as a single synergistic group. There are differences in muscle properties among the medial and lateral gastrocnemius (MG and LG, respectively) and the soleus that may affect their activation. Twelve volunteers participated in an investigation to determine whether the activation of the ankle plantarflexor muscles was modulated according to perturbation direction during unilateral standing perturbations of 1% body mass. High-density surface electromyography (HDS-EMG) was used to determine the amplitude and barycenter of the muscle activation and kinematic analysis was used to evaluate ankle, knee, and hip joint movement. The HDS-EMG amplitude and barycenter of MG and LG were modulated with the perturbation direction (MG p < 0.05; LG p < 0.01; one-way repeated-measures ANOVA). In soleus, the HDS-EMG barycenter modulated across the perturbation direction (p < 0.01 for X&Y coordinates), but the HDS-EMG amplitude did not change. A repeated-measures correlation was used to interpret the HDS-EMG pattern in the context of the kinematics. The relative contribution of MG activation compared to the total gastrocnemii activation was significantly associated with ankle dorsi/plantarflexion (rrm = 0.620), knee flexion/extension and abduction/adduction (rrm = 0.622 and rrm = 0.547, respectively), and hip flexion/extension and abduction/adduction (rrm = 0.653 and rrm = 0.432, respectively). The findings suggest that the central nervous system activates motor units within different regions of MG, LG and SOL in response to standing perturbations in different directions.

Effect of Synthetic Organoselenium Drug on the Degree of Pathological Changes in the Organs of White Mice Immunized with Tularemia and Brucellosis Vaccines.

We studied the effect of the organoselenium compound 2,6-dipyridinium-9-selenium-bicyclo[ 3,3,1]nonan dibromide (974zh) on the severity of pathological changes in the organs of experimental animals immunized with live tularemia and brucellosis vaccines. It was found that 974zh reduced reactogenicity of vaccines for experimental animals. Our findings indicate the prospects for further studies of the effects of 974zh on the functional state of experimental animals.

Lipid Fatty Acids from the Pericarp of Cydonia oblonga Mill. and Mespilus germanica L. are Involved in Plant Adaptation to Altitudinal Zonality.

The composition of fatty acids of total lipids of the outer and parenchymal parts of the pericarp in Cydonia oblonga Mill. and Mespilus germanica L. (Maloideae, Rosaceae), growing in the Northern Caucasian mountains at altitudes of 300, 500, 700, and 1200 m above sea level in various natural zones from experimental sites, was studied for the first time. It is established that the altitude of plant growth is largely correlated with the changes in the FA composition of the outer, but not the parenchymal, part of the pericarp. The nature of this variability suggests that the adaptation of plants to the conditions of significant temperature differences in the mountains is associated with the regulation of cell membrane fluidity, based on the interaction of opposite processes of synthesis of polyunsaturated and very-long-chain fatty acids.

Drug with Neuroprotective Properties Noopept Does Not Stimulate Cell Proliferation.

Effects of Noopept (N-phenylacetyl-L-prolylglycine ethyl ester) on the relative level of proliferation marker Ki-67 and cell cycle parameters were studied in HEK293 and SH-SY5Y cell lines. The previously established multifactorial mechanism of action of the drug includes enhancement of neurotrophin NGF and BDNF expression and increase in HIF-1 activity. The possible mitogenic action of Noopept was estimated by its effect on cell proliferation. Noopept did not affect cell distribution over G1, S, G2 cell cycle phases and the relative level of proliferation marker Ki-67 in the cell lines under study. These data suggest that Noopept does not stimulate cell growth.

The Unique Spectrum of Mutations in Patients with Hereditary Tyrosinemia Type 1 in Different Regions of the Russian Federation.

BACKGROUND: Hereditary tyrosinemia (HT1) is an autosomal recessive disorder characterized by impaired tyrosine catabolism because of fumarylacetoacetate hydrolase deficiency. HT1 is caused by homozygous or compound heterozygous mutations in the FAH gene. The HT1 frequency worldwide is 1:100,000-1:120,000 live births. The frequency of HT1 in the Russian Federation is unknown. AIM: To estimate the spectrum of mutations in HT1 in several ethnic groups of the Russian Federation. MATERIALS AND METHODS: From 2004 to 2017, 43 patients were diagnosed with HT1. The analysis of amino acids and succinylacetone was performed using NeoGram Amino Acids and Acylcarnitines Tandem Mass Spectrometry Kit and a Sciex QTrap 3200 quadrupole tandem mass spectrometer. Bi-directional DNA sequence analysis was performed on PCR products using an ABI Prism 3500. RESULTS: In the Russian Federation, the most common mutation associated with HT1 (32.5% of all mutant alleles) is c.1025C>T (p.Pro342Leu), which is typical for the Chechen ethnic group. Patients of the Yakut, the Buryat, and the Nenets origins had a homozygous mutation c.1090G>C (p.Glu364Gln). High frequency of these ethnicity-specific mutations is most likely due to the founder effect. In patients from Central Russia, the splicing site mutations c.554-1G>T and c.1062+5G>A were the most prevalent, which is similar to the data obtained in the Eastern and Central Europe countries. CONCLUSION: There are ethnic specificities in the spectrum of mutations in the FAH gene in HT1. The Chechen Republic has one of the highest prevalence of HT1 in the world.

[Early Markers of Cardiovascular Risk in Women With Hereditary Burdening by Cardiovascular Diseases].

PURPOSE: to identify early markers of development of cardiovascular diseases (CVD) in women. MATERIALS AND METHODS: Female firstdegree relatives from 39 families formed 2 groups: families (n=19) containing mothers with arterial hypertension (AH) (group 1) and healthy daughters (group 1a); families (n=20) containing practically heathy mothers (group 2) and healthy daughters (group 2a). We assessed data of anamnesis, including registration of cardiovascular risk factors, and family history of CVD. Examination included registration of anthropometric parameters, automatic and manual measurement of intima-media thickness (IMT) and resistance indexes of brachiocephalic arteries (BCA). We also determined cardio-ankle vascular index (CAVI), ankle-brachial index (ABI), and measured magnitude of ß-adrenoreception of membranes (ß-ARM) of erythrocytes in micro-quantities of venous blood. RESULTS: Mothers in both groups of families had excessive body mass or obesity. Mothers of group 1 had more pronounced signs of abdominal obesity (AO). They also had abnormalities of IMT and sings of subclinical atherosclerosis of BCA. CAVI in this group was significantly higher than in group 2. In group 1a median BMI (25.5 kg/m2) and waist/hip ratio were significantly higher than in group 2a. Daughters of group 1a contrary to group 2a had abnormalities of vascular wall: increased automatically measured IMT of carotid arteries and elevated CAVI. Arterial pressure and heart rate (HR) in group 1a were within limits of physiological norm but significantly higher than in group 2a. All included women had elevated ß-ARM values but in group 1a this parameter was significantly higher than in group 2a and moderately correlated with HR. CONCLUSIONS: Risk factors of CVD development in women are AH, AO, high activity of the sympathoadrenal system. These factors provoke changes of vascular wall (elevation of its stiffness and early subclinical atherosclerosis). In daughters of mothers with AH important prognostic components of CVD risk in addition to family history of CVD are AO, systolic blood pressure (BP) >120 mm Hg, diastolic BP >78 mm Hg, HR approaching upper limit of physiological norm, and high CAVI (indicator of vascular wall stiffness).

[The role of changes in the structure of water and water-based systems in the mechanism of combined action of the magnetic field and natural therapeutic factors].

The application of the dielectrometric technology in the combination with the resonance method and thermometry has been shown to provide an insight into a variety of factors including the influence of the magnetic field of the permanent magnets on the structure of water and water-containing systems. It was found that the magnetization of both distilled water and «Karachinskaya¼ mineral water increase their structuredness. Magnetization changes the biological properties of therapeutic peloids by virtue of a change in the structure of water and, accordingly, the structuring of the water fraction of therapeutic muds. The analysis of the results of experimental and theoretical studies gives evidence that variations in the structure of water and water-based systems play an important role in enhancing the effectiveness of the combined treatment with the use of the magnetic field and the natural therapeutic factors. The positive and adaptogenic effects of such treatment appears to be due to such changes in the structure of water and water-containing systems which facilitate communication between the processes in the resonant mode but maintain stability of the cellular structures by limiting the mobility of water dipoles under the influence of non-resonant frequencies.