Transcriptome analysis reveals multiple targets of erythritol-related transcription factor EUF1 in unconventional yeast Yarrowia Lipolytica.

BACKGROUND: Erythritol is a four-carbon polyol with an unclear role in metabolism of some unconventional yeasts. Its production has been linked to the osmotic stress response, but the mechanism of stress protection remains unclear. Additionally, erythritol can be used as a carbon source. In the yeast Yarrowia lipolytica, its assimilation is activated by the transcription factor Euf1. The study investigates whether this factor can link erythritol to other processes in the cell. RESULTS: The research was performed on two closely related strains of Y. lipolytica: MK1 and K1, where strain K1 has no functional Euf1. Cultures were carried out in erythritol-containing and erythritol-free media. Transcriptome analysis revealed the effect of Euf1 on the regulation of more than 150 genes. Some of these could be easily connected with different aspects of erythritol assimilation, such as: utilization pathway, a new potential isoform of transketolase, or polyol transporters. However, many of the upregulated genes have never been linked to metabolism of erythritol. The most prominent examples are the degradation pathway of branched-chain amino acids and the glyoxylate cycle. The high transcription of genes affected by Euf1 is still dependent on the erythritol concentration in the medium. Moreover, almost all up-regulated genes have an ATGCA motif in the promoter sequence. CONCLUSIONS: These findings may be particularly relevant given the increasing use of erythritol-induced promoters in genetic engineering of Y. lipolytica. Moreover, use of this yeast in biotechnological processes often takes place under osmotic stress conditions. Erythritol might be produce as a by-product, thus better understanding of its influence on cell metabolism could facilitate processes optimization.

[Clinical efficiency of occlusive splints in the treatment of patients with temporomandibular joint dysfunction complicated by bruxism]. / Klinicheskaya effektivnost' okklyuzionnykh shin v lechenii patsientov s disfunktsiei visochno-nizhnechelyustnogo sustava, oslozhnennoi bruksizmom.

OBJECTIVE: Improving the effectiveness of treatment of patients with temporomandibular joint (TMJ) dysfunction complicated by bruxism. MATERIAL AND METHODS: 70 patients with signs of parafunction of the masticatory muscles were selected for the clinical study. The patients were divided into 2 groups of 35 people. Splints were made for patients from the groups using 2 different 3D printing methods. The treatment was monitored using clinical and instrumental studies, electromyography of the muscles themselves and computed tomography of the TMJ after 3, 6 and 12 months. RESULTS: The severity of hypertonicity of the masticatory muscles decreased by the end of 3 months of treatment with the help of repositioning occlusal splints. After 6 months of treatment, the mean BEA decreased in both study groups, but to a lesser extent than in the first 3 months. After wearing a splint after 12 months, the decrease in muscle tone was insignificant. CONCLUSION: The clinical use of positioning splints plays a crucial role in the stabilization of occlusion and is a mandatory stage of complex treatment of patients with pathology of the occlusal-musculoskeletal complex.

Compact liquid analyzer based on a resonator with a lateral excitation electric field.

A compact hardware-software complex for measuring the electrical and mechanical parameters of a liquid based on a piezoelectric resonator with a lateral exciting electric field has been developed. It is shown that the modulus of the electrical impedance of the resonator near the resonant frequency decreases monotonically with an increase in conductivity of the contacting liquid. The obtained dependences can be used as calibration curves for measuring conductivity in the range of 100-10,000 µS/cm. A method for the simultaneous determination of the modulus of elasticity, viscosity, and permittivity of a liquid using the electromechanical equivalent scheme was developed and tested on a mixture "water - glycerol" with the different glycerol content.

Microbial acoustic sensor test-system based on a piezoelectric resonator with a lateral electric field for kanamycin detection in liquid.

A microbial test-system for real-time determination of low/residual concentrations of kanamycin in a liquid without the need for special labels is presented. The main element of the system was a piezoelectric resonator excited by a lateral electric field based on an X-cut lithium niobate plate 0.5 mm thick with two rectangular electrodes on one side. On the other side of the resonator, there was a 1.5 ml liquid container. As a sensory element we used Escherichia coli B-878 microbial cells, which are sensitive to kanamycin. For measurement 1 ml of this cells suspension was placed in a liquid container and then the test liquid in the amount of 2 µl containing kanamycin was added. The change in the real part of the electrical impedance of the resonator before and after the test liquid addition was used as an analytical signal which indicated the presence of kanamycin. The lower limit of determination of kanamycin turned out to be 1.0 µg/ml with an analysis time of 10 min. The test-system allows to detect kanamycin in the presence of such antibiotic as ampicillin and polymixin.

Newborns from women infected with COVID-19: somatic and metabolic status.

To date, there are limited data regarding manifestations of new coronavirus infection in infants born of SARS-CoV-2 infected mothers, so the aim of this study is to investigate somatic and metabolic status of newborn infants born to mothers diagnosed with COVID-19. The investigation was carried out on the bases of Laboratory Diagnostic Department of Samara Regional Clinical Hospital named after V.D. Seredavin and the Department of Fundamental and Clinical Biochemistry with Laboratory Diagnostics of Samara State Medical University. Under observation were 85 newborns, including 35 born of healthy mothers and 50 born of COVID-19 mothers.The somatic status of all newborns was assessed using the Apgar scale at the 1st and 5th minutes after birth. Also all newborns had general and biochemical blood tests and newborns from mothers with COVID-19 were tested for the presence of SARS-CoV-2 RNA in oral and nasopharyngeal swabs. Thus, the study of somatic status revealed that of 50 neonates from women infected with COVID-19, only 18% were practically healthy, the rest had signs of prematurity, hypotrophy, perinatal CNS damage, diabetic fetopathy, pulmonary atelectasis, delayed intrauterine development, asphyxia. The metabolic state is characterised by decreased haemoglobin and platelets, increased concentration of total protein, including C-reactive protein, high transaminase activity, decreased sodium and chloride content. These parameters of general and biochemical blood tests can be considered as indicators for the evaluation of the condition of newborns from mothers with COVID-19.

Molecular profile of oral fluid in new coronavirus infection.

Oral fluid is an alternative biological material that confirms correlations with blood parameters in various pathological conditions of the body. In order to find a non-invasive approach to stratification of patients with COVID-19 disease, molecular biomarkers of the oral fluid have been determined in patients with moderate coronavirus infection in comparison with clinically healthy individuals. It has been shown that proteomic, carbohydrate, macro- and microelement profiles of the oral fluid in coronavirus infection can be used for diagnostics. The features of protein metabolism were revealed: an increase in the content of total protein, urea; increased activity of enzymes aspartate aminotransferase, gamma glutamyl transpeptidase, creatine phosphokinase, alkaline phosphatase; changes in carbohydrate metabolism, which is expressed by an increase in glucose and lactate levels, an increase in lactate dehydrogenase activity, sodium, chloride, calcium, magnesium, iron content.

[Biomarkers of system inflammation in local and diffuse peritonitis]. / Markery sistemnogo vospaleniia pri mestnom i rasprostranennom peritonite.

In cases of any acute surgical abdominal disease the progression of purulent inflammation can lead to local or diffuse peritonitis. The indicators of the degree and specificity of the inflammatory response in blood such as cytokine concentration, neutrophil activity, plasma antioxidant capacity (thiols concentration) could be considered as potential predictors of complications. The luminol-dependent chemiluminescence (CL) response of blood activated by the phorbol ester (PMA), and the concentration of cytokines IL-6, IL-8, IL-10, myeloperoxidase (MPO) and thiols in plasma were measured in patients with uncomplicated condition (group 1, n=8), local peritonitis (group 2, n=9) or diffuse peritonitis (group 3, n=9) at admission to surgery (before surgical operation, b/o), immediately after surgical operation (a/o) and a day after surgery (1 day) as well as in healthy volunteers (norm, n=12). In all time-points the cytokines and MPO concentrations measured by ELISA, in group 3 were higher than in healthy volunteers and in patients in groups 1 and 2. Blood CL demonstrated a more than 5-fold increase above the normal values in all patients, and was also higher in group 2 as compared to group 1 (b/o and a/o). Patients in group 3 had shown both maximum and minimum of CL values, which could be a consequence of neutrophil priming or exhaustion ("immune paralysis"), respectively. The same patients' plasma exhibited low thiol concentration (≤30% vs normal values). In patients with fatal outcomes (group 3, n=2) within a day after surgery, either a decrease of the CL to zero values concurrently with elevated IL-8 and IL-6 concentrations and low thiol levels was observed, or CL exceeded normal values more than 20 times with concurrent complete exhaustion of the plasma thiol pool. No clear dependency between the plasma parameters and neutrophil activity was found. Hence a parameter set for prognosis and/or early diagnosis of infectious complications in acute abdominal pathology should include different biomarkers of the inflammatory response: cytokine profile (IL-6, IL-8, IL-10), MPO and neutrophil activity, antioxidant plasma capacity (e.g., total thiols concentration).

[Takayusu Arteritis on the background of congenital vascular anomaly?]

The article described a female patient with high-grade arterial hypertension diagnosed at her young age. Congenital abdominal aortic hyperplasia and conformity with criteria for Takayasu arteritis were diagnosed at 10 years following the clinical manifestation.

Metabolic engineering of Saccharomyces cerevisiae for the de novo production of psilocybin and related tryptamine derivatives.

Psilocybin is a tryptamine-derived psychoactive alkaloid found mainly in the fungal genus Psilocybe, among others, and is the active ingredient in so-called "magic mushrooms". Although its notoriety originates from its psychotropic properties and popular use as a recreational drug, clinical trials have recently recognized psilocybin as a promising candidate for the treatment of various psychological and neurological afflictions. In this work, we demonstrate the de novo biosynthetic production of psilocybin and related tryptamine derivatives in Saccharomyces cerevisiae by expression of a heterologous biosynthesis pathway sourced from Psilocybe cubensis. Additionally, we achieve improved product titers by supplementing the pathway with a novel cytochrome P450 reductase from P. cubensis. Further rational engineering resulted in a final production strain producing 627 ± 140 mg/L of psilocybin and 580 ± 276 mg/L of the dephosphorylated degradation product psilocin in triplicate controlled fed-batch fermentations in minimal synthetic media. Pathway intermediates baeocystin, nor norbaeocystin as well the dephosphorylated baeocystin degradation product norpsilocin were also detected in strains engineered for psilocybin production. We also demonstrate the biosynthetic production of natural tryptamine derivative aeruginascin as well as the production of a new-to-nature tryptamine derivative N-acetyl-4-hydroxytryptamine. These results lay the foundation for the biotechnological production of psilocybin in a controlled environment for pharmaceutical applications, and provide a starting point for the biosynthetic production of other tryptamine derivatives of therapeutic relevance.

[Blood group and human diseases (review of literature).]

AB0 blood group antigens were discovered over a century ago; however, it is still important to study their role in development of various pathological conditions. Today it is known that antigenic determinants of this blood group are present not only on erythrocyte membrane but also on other cells and tissues: platelets, gastrointestinal epithelium and salivary glands, respiratory system cells. In the last decade, a large number of studies have appeared to reveal the relationship between a specific disease and blood group type, meta-analyses have been published. Previously, the authors have studied the metabolic status, cell composition and coagulation profile of clinically healthy individuals for more than on 180,000 donations, that allowed to identify groupspecific features for each blood group. This review presents generalized data on the association of such pathological conditions as coronary heart disease, thromboembolic complications, tumors of various localizations, inflammatory and destructive oral diseases, psychiatric and some infectious diseases with the presence or absence of antigenic determinants A and B. Carriers of blood group 0 (I) are generally more resistant to diseases, with the exception of H.pylori-associated gastrointestinal diseases. Carriers of «antigenic¼ blood groups A (II), B (III), AB (IV) are more susceptible to development of infectious, cardiovascular and cancer diseases. The presented data demonstrate clinical significance of the definition of group typing not only for selection of blood and its components during transfusion and transplantation, but also for diagnostics, determination of risk group and tactics for treatment patients with different nosologies.

[Сlinicmolecular indicators of inflammatory destructive damage of the oral cavity in periodontitis in persons with various group accessories of blood.]

In order to find a connection between the alteration of oral tissues and genetic predisposition to inflammatory and destructive processes in oral media, the cytokine profile of the oral fluid of clinically healthy individuals was determined for various blood group affiliations according to the AB0 system. The group-specific features of individuals with B(III) blood group were revealed: an increase of 32,5% in the content of interleukin-6 and 63,1% in the content of interleukin-8 compared with similar data for people with 0(I), A(II), AB(IV) blood groups, which can predispose to the greatest activity of the inflammatory process in the oral cavity in individuals with antigen B. Confirmation of this fact is an increase of IgA antibodies to gliadin in the blood among patients with chronic generalized periodontitis with B(III) blood group, up to 5,00 U/ml (p<0,01), which indicates the processes of acute inflammation, and along with an increase in blood IgG antibodies to transglutaminase in comparison with a group of clinically healthy individuals, it serves as an indicator of damage to the body's connective tissue at the molecular level. When examining the dental status, pronounced clinical manifestations of chronic generalized periodontitis were found in patients with A(II) blood group, the molecular foundation of which is the highest content of IgA and IgG antibodies to transglutaminase in the oral fluid (0,35 U/ml and 0,45 U/ml), which contributes to the activation of periodontal-destroying inflammatory and inflammatory processes, obviously, with a tendency to the chronic course of the disease. The studies performed allowed us to analyze in clinically healthy individuals a predisposition to alternative processes in oral environments, using gradation by group blood affiliation.

Influence of the conductivity of a liquid contacting with a lateral electric field excited resonator based on PZT ceramics on its characteristics.

The influence of the conductivity of a liquid contacting with a piezoelectric resonator with a lateral electric field based on a PZT piezoceramic plate was experimentally and theoretically investigated. In this resonator the shear component of the mechanical displacement, which does not lead to radiation losses upon contact with the liquid, was prevalent. The frequency dependences of the real and imaginary parts of the electrical impedance of the resonator in the frequency range 50-300 kHz showed the presence of three resonances at frequencies of 68.7, 97.8 and 264 kHz with the values of the electro-mechanical coupling coefficient of 12.2%, 14.7% and 6.5%, respectively. The quality factor of each resonance in contact with the liquid turned out to be significantly higher than the quality factor of the resonator with a longitudinal acoustic wave based on lithium niobate. The dependences of the maximum value of the real part of the electrical impedance of such a resonator on the conductivity of the liquid for each resonance were obtained. This experimental data turned out to be in a good agreement with theoretical results obtained by using the method of equivalent circuit.

A teaching protocol demonstrating the use of EasyClone and CRISPR/Cas9 for metabolic engineering of Saccharomyces cerevisiae and Yarrowia lipolytica.

We present a teaching protocol suitable for demonstrating the use of EasyClone and CRISPR/Cas9 for metabolic engineering of industrially relevant yeasts Saccharomyces cerevisiae and Yarrowia lipolytica, using ß-carotene production as a case study. The protocol details all steps required to generate DNA parts, transform and genotype yeast, and perform a phenotypic screen to determine ß-carotene production. The protocol is intended to be used as an instruction manual for a two-week practical course aimed at M.Sc. and Ph.D. students. The protocol details all necessary steps for students to engineer yeast to produce ß-carotene and serves as a practical introduction to the principles of metabolic engineering including the concepts of boosting native precursor supply and alleviating rate-limiting steps. It also highlights key differences in the metabolism and heterologous production capacity of two industrially relevant yeast species. The protocol is divided into daily experiments covering a two-week period and provides detailed instructions for every step meaning this protocol can be used 'as is' for a teaching course or as a case study for how yeast can be engineered to produce value-added molecules.

New approach for determination of antimicrobial susceptibility to antibiotics by an acoustic sensor.

For the first time, a rapid method was proposed to determine the susceptibility of Escherichia coli cells to antibiotics by the example of ampicillin by using a biological sensor based on a slot mode in an acoustic delay line. It has been established that an indicator of the antibiotic activity to microbial cells is the difference between the recorded sensor's signal before and after exposure cells with antibiotic. The depth and frequency of the peaks of resonant absorption in the frequency dependence of the insertion loss of sensor varied after adding an antibiotic with different concentrations to the microbial cells. By using the acoustic sensor based on slot-mode a criterion of E. coli sensitivity to ampicillin was established. The advantages of this method are the ability to carry out the analysis directly in the liquid, the short analysis time (within 10-15 min), and the possibility to reusable sensor.

The sensor for measuring the micro-displacements based on the piezoelectric resonator with lateral electric field.

Theoretical and experimental studies of the influence of a thin metal film on the characteristics of a piezoelectric resonator with a lateral electric field showed the possibility of creating a micro-displacement meter in the range of 10-300 µm. For the experiments, two resonators based on the plates of PZT piezoceramics with a thickness of 3.56 and 4.46 mm with resonant frequencies of ~96 and ~260 kHz for both resonators were used. It has been experimentally established that in both cases, with an increase in the width of the gap between the free side of the piezoelectric resonator and thin aluminum film, the frequency of the parallel resonance and maximum value of the real part of the electrical impedance increase and reach saturation. Besides, it has been shown that the relative change of these values with a change in the width of the gap in the range 10-300 µm increases with decreasing the thickness of the resonator. In this case, the frequency of the series resonance practically does not change. It has been also established that the sensitivity of the resonator to the presence of a conducting film at the resonant frequency of ~96 kHz is significantly higher in comparison with the resonant frequency of ~260 kHz. The experimental results are in the qualitative agreement with the theoretical data.

[Coagulation test features depending on the AB0-blood groups system antigenic composition.]

The maintenance of normal blood flow through the vessels is the result of the coordinated work of the coagulation and anticoagulation systems of our body. The balance of this system depends on many factors, including endothelial, humoral, platelet ones, however, we still lack knowledge about the effect of antigenic determinants on the state of the hemostatic system. This study is devoted to assessing the effect of the presence and absence of antigens on the AB0 system, presented on erythrocyte and platelet membranes, on hemostatic parameters. The study was conducted in the Clinics of Samara State Medical University and consisted of127 clinically healthy individuals who underwent a general analysis and biochemical blood analysis, 52 people with the most stable indicators of cell composition and metabolic profile were selected for a coagulation test, including determination of the activity of coagulation factors and routine tests. A significant decrease in the activity of the VIII and VII coagulation factors was revealed, as well as an increase in the prothrombin time in patients with 0 (I) blood group compared to the "antigenic" blood groups. The presence of biological variation for indicators of external and internal coagulation paths was noted, depending on the group of blood belonging to the AB0 system. The findings suggest that there is an increased susceptibility to bleeding in patients with 0 (I) blood groups due to the absence of antigenic determinants on the cell membrane, while for "antigenic" blood groups, on the contrary, there is a susceptibility to thrombosis due to increased activity of the components of the coagulation system.

Sensor for ampicillin based on a microwave electrodynamic resonator.

The paper describes a new biological sensor which represents a resonator based on a segment of a rectangular waveguide of 8 GHz band with shear dimensions of 28.5 × 12.6 mm2. On one side, the resonator is bounded by a metallic short-circuited wall; on the other side, it is bounded by a lithium niobate plate with a porous polystyrene film. This film, applied by centrifugation and modified in high-frequency discharge plasma in argon, was used to immobilize cells of Escherichia coli K-12. This resonator was connected through a coaxial-waveguide adapter to the S parameter meter, by means of which the reflection coefficient S11 in the plane of the lithium niobate plate was measured. The addition of an aqueous solution of ampicillin at 4-50 µg/ml to immobilized cells led to a significant change in the reflection coefficient of S11 from - 10.15 dB to - 15.09 dB. At the same time, the resonance frequency changed insignificantly within the range 8.06-8.068 GHz. The optimal time for modifying the polystyrene film for obtaining the required porosity and the optimal time for the immobilization of the bacterial cells were determined. The immobilized cells retained their activity for 4 months at a temperature of 4 °C. The study showed the promise of such a biosensor to determine ß-lactam antibiotics in aqueous solutions by using ampicillin as an example. The limit of detection of the developed biosensor with respect to ampicillin was established (4 µg/ml).

Effect of the conductivity of a thin film located near the acoustic delay line on the characteristics of propagating SH0 wave.

The results of the study of the influence of the conductivity of a thin film located with a certain gap above the acoustic delay line on the characteristics of propagating acoustic wave with a quasi-shear-horizontal polarization (SH0) are presented. The delay line was made of Y-X lithium niobate plate with a thickness of 200 µm containing two interdigital transducers for exciting and receiving SH0 wave. The operating frequency range of the delay line was equal to 2.6-3.8 MHz. The 1 mm thick glass plate with a film of the given conductivity applied on one plate's side was placed above this delay line with the some air gap. The change in the phase of output signal of the delay line was measured at the various values of the width of this gap. As films with the various values of the conductivity, tin dioxide films deposited on a glass plate by high-frequency reactive magnetron sputtering in the oxygen-argon mixture were used. It has been shown that the velocity of SH0 wave propagating in the delay line under investigation depends on the film conductivity and the width of the gap between the delay line and conductive film. The dependences of the change in the SH0 wave velocity on a width of the gap between the surface of the delay line and conductive film were constructed, as well as on the film conductivity. The comparison of the experimental results with the theoretical data showed their good agreement.

The effect of the conductivity of a film located near a piezoelectric resonator with a lateral electric field based on the PZT ceramics on its characteristics.

In this paper, we investigated theoretically and experimentally the effect of the conductivity of the film located in the immediate vicinity of the free side of a piezoelectric resonator based on the PZT ceramics on its characteristics. It is shown that with increasing the conductivity of the film at first the parallel resonant frequency does not change, then it decreases in a certain interval and after that remains practically constant. With increasing the width of the gap between the film and the resonator, the resonant frequency increases and reaches the saturation. At that the degree of the total frequency change decreases with decreasing the conductivity of the film. We also investigated the dependence of the maximum value of the real part of the electrical impedance on the film conductivity at the different values of the width of the gap between the film and the resonator. The dependences of the maximum value of the real part of the electrical impedance on the width of the gap between the film and the resonator are also obtained for different values of the film conductivity. A comparison of the theoretical and experimental data showed their good qualitative agreement.

[The use of the visuo-motor reaction training device for the improvement of the coordination in the eye-hand system of the children and adolescents following the completion of the antineoplastic treatment of brain tumours]. / Primenenie trenazhera zritel'no-motornoi korrektsii dlia uluchsheniia koordinatsii v sisteme glaz-ruka u detei i podrostkov posle zaversheniia protivoopukholevoi terapii novoobrazovanii golovnogo mozga.

The survivors among the children and adolescents with a brain tumour are likely to show evidence of the impairment of the most important cognitive functions, such as attention, visual-motor integration, and working memory, following the completion of the antineoplastic treatment. The basic characteristics of these functions compromised in the patients presenting with the most serious cognitive deficiency are the information processing rate and the time needed for carrying out any cognitive activity in the patients experiencing deficit of white matter in the brain. AIM: The objective of the present study was to demonstrate the possibility and the effectiveness of the application of the new method for the stimulation of the information processing activity of the brain, the enhancement of its effectiveness, and the improvement of the quality of the visual-motor coordination in the children and adolescents following the completion of the antineoplastic treatment of brain tumours. This paper reports the first experience of the application of the Dynavision d2 visuo-motor reaction training device which has been used for the visual-motor integration and the increase of the information processing speed. MATERIAL AND METHODS: The sample included in the study was comprised of a total of 46 children at the mean age of 10.6±3.34 years that had undergone the treatment of the brain neoplasm. Each patient participated in 5 to 8 sessions of exercises with the use of the the Dynavision d2 visuo-motor reaction training device during 21 days. RESULTS: All the treated patients exhibited the stable reduction of the mean time of the reaction from the first session to the last one. The comparison of the outcomes of the treatment between the children of different sexes has demonstrated better results (in absolute terms and in dynamics) in the girls in comparison with the boys. The exercises with the use of the Dynavision d2 visuo-motor reaction training device were not accompanied by any negative subjective sensations in the treated patients, nor did they induce the adverse somatic side effects. CONCLUSION: The results of the present study give evidence of both the applicability and the effectiveness of the application of the Dynavision d2 visuo-motor reaction training device for the diagnostics of the disturbances in the motor and visual coordination and the associated cognitive functions and for their correction in the children following the treatment of the neoplasm of the posterior cranial fossa.