Characterization of CoCas9 nuclease from Capnocytophaga ochracea.

Cas9 nucleases are widely used for genome editing and engineering. Cas9 enzymes encoded by CRISPR-Cas defence systems of various prokaryotic organisms possess different properties such as target site preferences, size, and DNA cleavage efficiency. Here, we biochemically characterized CoCas9 from Capnocytophaga ochracea, a bacterium that inhabits the oral cavity of humans and contributes to plaque formation on teeth. CoCas9 recognizes a novel 5'-NRRWC-3' PAM and efficiently cleaves DNA in vitro. Functional characterization of CoCas9 opens ways for genetic engineering of C. ochracea using its endogenous CRISPR-Cas system. The novel PAM requirement makes CoCas9 potentially useful in genome editing applications.

Effects of Combined Exposure to Carbon Ions and Protons on the Pool of MCF-7 Breast Cancer Stem Cells In Vitro.

We studied the effects of single and combined action of protons and carbon ions 12C6+ on the pool of MCF-7 human breast cancer stem cells. Single irradiation with a beam of protons or carbon ions had no significant effects on the relative number of cancer stem cells (CSC). The effects of combined irradiation in a total equieffective dose of 4 Gy depended on the sequence of exposure to ionizing radiations: the relative number of CSC did not change after irradiation with carbon ions and then with protons, but increased in the case of the reverse sequence. The most favorable result, i.e. a decrease in the CSC pool, was observed in the case of sequential irradiation with carbon ions and protons and their equal contribution to total equieffective dose. In this case, the absolute number of CSC decreased by on average 2.1 times in comparison with the control (p<0.05). The revealed regularities are of interest for the further development of new methods of radiation therapy.

EVALUATION OF CHANGES IN THE PROFILE OF THE FACE DURING ORTHODONTIC TREATMENT OF DISTAL OCCLUSION CAUSED BY ANTEPOSITION OF THE UPPER JAW.

Distal occlusion of the dentition is one of the most common anomalies in the Eastern European population. The widespread use of conservative treatment methods creates the need to predict the results of tooth movement. The problem of the influence of orthodontic treatment on the configuration of the soft tissues of the face is of paramount importance for the orthodontist. Purpose of the study: to evaluate changes in the profile of the face during orthodontic treatment of distal occlusion due to anteposition of the upper jaw. 13 patients aged 18-30 years with distal occlusion caused by anteposition of the upper jaw were examined and treated. A standard examination was carried out, including clinical, anthropometric and radiological diagnostic methods. Orthodontic treatment of all patients was carried out, including the removal of individual teeth in the upper jaw. The study shows the features of changing the configuration of the soft tissue profile of the face in the treatment of patients with distal occlusion of the dentition. As a result of orthodontic treatment, the position of the jaws in the sagittal and vertical planes remained practically unchanged. The soft tissue profile of the face was affected by a change in the position of the incisors. A thorough diagnosis of dentoalveolar anomalies, identification of facial configuration features and knowledge of the changes that accompany orthodontic movements will lead not only to the normalization of occlusion and function, but also to the harmonization of the soft tissues of the face.

DISEASES OF THE ORAL MUCOSA IN PATIENTS IN THE POST-COVID PERIOD.

The impact of SARS-CoV-2 (severe acute respiratory syndrome-related coronavirus-2) on the entire human body causes irreversible changes in all organs and systems. Complications in the form of chronic diseases require the treatment of clinicians in various fields of medicine. Dentists are faced with diseases of the oral mucosa in apparently healthy patients who are not predisposed to them but have undergone COVID-19. Purpose of the study - to assess the impact of the coronavirus infection (COVID-19) on the course of diseases of the oral mucosa. We examined 51 patients aged 20 to 75 who had undergone COVID-19 with various diseases of the oral mucosa. The study used clinical and laboratory methods. The features of the clinical course of certain diseases of the oral mucosa were revealed, the dynamics of the local immune defense of the oral cavity in the post-covid period was determined.

[Analysis of the quality of orthodontic care for children with partial defects of dentition in the dental organizations of various forms of ownership]. / Analiz kachestva ortodonticheskoi pomoshchi detyam pri chastichnykh defektakh zubnykh ryadov v stomatologicheskikh organizatsiyakh razlichnoi formy sobstvennosti.

THE AIM OF THE STUDY WAS: To conduct a comparative analysis of the quality of medical care for children and adolescents with dentoalveolar anomalies in dental medical organizations of different organizational and legal basis and forms of ownership. MATERIAL AND METHODS: The analysis of orthodontic records of children and adolescents with dentoalveolar anomalies and factors predisposing to their development in 1032 patients aged from 6 months to 18 years was carried out with the use of orthodontic patient records classification of L.S. Persin (2020). The effectiveness of orthodontic care for children and adolescents with various forms of anomalies of the dentition was analyzed. The draft clinical guidelines have been developed depending on the patient model. RESULTS: As shown by the data of orthodontic records of patients, 12% of patients applied during the formation of occlusion of deciduous teeth, 9.2% of children in the early mixed dentition phase, 66.5% of patients in the late mixed dentition phase, and 12.2% of patients first admitted in the permanent dentition phase. As for the analysis of the quality of orthodontic care for children and adolescents with dentoalveolar anomalies in dental organizations of various forms of ownership, the results are controversial. CONCLUSION: Most often in the orthodontic charts data on dentition anomalies were filled, followed by the anomalies of the teeth, anomalies of occlusion in the sagittal direction, anomalies of occlusion in the vertical direction and partial defects of the dentition. Anomalies of soft tissue attachment, anomalies of occlusion of molars on the right and left with palatoocclusion, and anomalies of occlusion of molars on the right and left with linguo-occlusion were less common in orthodontic charts.

BIOMECHANICS OF CHANGING THE POSITION OF PERMANENT TEETH WITH EARLY LOSS OF THE FIRST TEMPORARY MOLARS.

Premature removal of baby teeth is one of the main causes of deformation of the dentition. The loss of the first temporal molars is most often reported (39.9±4.8%), which leads to displacement of the teeth bordering on the defect, narrowing and shortening of the dental arch, which then leads to retention of premolars, cutting out permanent canines outside the dental arch, pathology of cutting out permanent molars. The aim of the study was to evaluate the biomechanics of permanent teeth migration in patients who prematurely lost their first temporal molars. An anthropometric study of jaw models and orthopantomograms was conducted in 25 patients (age 7-12 years) who had premature loss of the first temporal molars of the upper and lower jaws (age 7-12 years). Complications were evaluated, regularity was revealed and the biomechanics of permanent teeth displacement as a result of the prolonged absence of temporary molars were determined.

CLINICAL AND RADIOGRAPHIC CHANGES FOLLOWING ORTHODONTIC INTRUSION OF OVERERUPTED MAXILLARY MOLARS WITH TWO MINI-IMPLANTS.

Objective - to evaluate clinical and radiographic changes of orthodontic intrusion of upper first molars with two mini-implants, using light intrusion forces, to create the space for prosthetic rehabilitation on the lower jaw. In 20 patients (aged between 26.8 and 45) with secondary deformities in the lateral region in the vertical direction, associated with partial absence of teeth in the lower jaw, and orthodontic preparation for subsequent prosthetics was performed. Each subject was missing 1-2 teeth in the lateral segment. All patients were divided into two groups: with intrusion mechanics with orthodontic implants (20 people) and intrusion with an orthodontic shape memory arch (20 people). The radiographic changes in male and female groups were assessed and compared based on the panoramic radiographs and CBCT data before and after intrusion. In the study 28 molars were fully intruded and their position was normalized with two mini-implants placed palatally and buccally. Mean extrusion time was 7.86±0.42 months (P<0.001), mean intrusion length was 2.97±0.15 mm (P<0.001), and mean change of mesial molar inclination was 2.02±0.44 degrees (P<0,005). The degree of root resorption was evaluated according to CBCT data in Multiplanar reconstruction (MPR); 50% of molars had grade 0 of resorption, 42.85% had grade 1, and 7.15% of molars had grade 2. There was no resorption in the trifurcation area. Intrusion of molars can be successfully accomplished with 2 mini-implants, placed palatally and buccally, with a light traction force (30-50 g) of an elastic chain.

[Implementation of the national program of cancer screening of oral precancerous and oncological diseases in the Russian Federation]. / Realizatsiia natsional'noi programmy onkoskrininga predrakovykh i onkologicheskikh zabolevanii slizistoi obolochki rta u naseleniia Rossiiskoi Federatsii.

The aim of the study is to implement the national project created within the framework of the state task of the Ministry of Education and Science of the Russian Federation 'Development of the program of screening of cancer and precancerous diseases of the oral mucosa and red border of the lips'. For its approbation and implementation, national research groups have been created at the bases of the Department of Therapeutic Dentistry of the Moscow State Medical and Dental University, the Tula regional dental clinic, City dental clinic in Noyabrsk City (Yamal-Nenets autonomous district), which in the period from 2015 to the first half of 2018 studied the epidemiology of cancer and precancer of the oral mucosa and the vermilion among 217 806 people aged 18-92 years. Over three and a half years, the effectiveness of the research team increased by 8.45 times. The results of the study confirmed the low oncological alertness of dentists. Educational working program for dentists training 'Screening of diagnostics of precancerous diseases and cancer of the oral mucosa - AFS-D Set' was developed by coordinators of the project and implemented for training of 574 dentists within continuous medical education programs.

[Ontogenetic aspects of changing lysozyme activity under acute somatic pain].

OBJECTIVE: Ouraim was to study influence of acute somatic pain on lysozyme activity of rats of different age: newborn, rats after eye opening, rats at the age of a month, adult and old rats. METHODS: Lysozyme activity was checked before pain irritation and 2, 30, 60, 120, 180min afterwards using Dorofeychuk's method in our modification. Pain effect was modeling by electrical stimulation. RESULTS: activity of lysozyme was 0. 434±0. 01 units in intact newborn rats, It was higher than in adult rats - 0.260 ±0.01 units (p <0.001) and it was unchanged during the experiment. We found low lysozyme activity in rats after eye opening - 0.015±0.003 units and it was stable during the experiment. Rats at the age ofa month had diphasic reaction: lysozyme activity was 0.191±0.01 units in intact rats, it increased up to 0.378±0.01 units (p <0.001) in 2 min after painful irritation and it decreased up to 0.113±0.02 units (p <0.001) in 30 min. Lysozyme activity was 0.260±0.01 units. Single-phase reaction was determined after acute painful irritation: increase of lysozyme activity after acute somatic pain up to 0,450±0,014 units (p <0.001). Lysozyme activity was 0.246±0.02 units in blood plasma of old rats. It decreased up to 0.1701±0.01 units (p <0.01) after painful irritation and it was 0.387±0.01 (p <0.001) in the end of the experiment. CONCLUSION: Response on pain irritation has differences in different groups. The common vector of pain response was the increase of lysozyme activity in rats at the age of a month, adult rats, rats after eye opening and old rats. Reaction of increasing lysozyme activity was strongly detected in adult rats. The results demonstrate preventive lasozyme resistance to potential tissue damage or contamination.

[Investigation of actual nutrition and some factors of physical status in rugby players].

Physical status and nutrition and energy value of factual diets of 18 rugby players (15-30 years old) assessed during current medical observation were analyzed. Anthropometric data (height, body mass--BM, waist circumference/ hip circumference index, body mass index--BMI), body composition data and resting metabolic rate by bioimpedance method (analyzer ABC-01 "Medass") were determined. These indices were found to be normal except more high level of BMI due to high levels of lean and skeletal muscle mass, but not a result of obesity. Estimation of factual nutrition based on 1-day feeding (nutrition-screening questionnaire method) has demonstrate that it was not optimal both for energy and nutrition value for this group of physical activity. Energy value ranged from 1889 kcal to 3175 kcal (22.5-35.1 kcal/kg BM/day) with lower carbohydrate intake (46.9%) and higher fat intake (35.8%) were found. Nutrition value ranges: protein--67-144 g (0.71-1.38 kcal/kg BM/day), fat--75-140 g (0.88-1.51 kcal/kg BM/day), carbohydrate--173-420 g (2.17-4.66 kcal/kg BM/day) were observed. The findings suggest the need to pay more attention to in-depth study of dietary intake of athletes to maintain health and to create optimal conditions for a professional sports activities.

[A longitudinal study of pediatric-onset multiple sclerosis in the Voronezh region]. / Prospektivnyi analiz klinicheskogo techeniya rasseyannogo skleroza s debyutom v detskom i podrostkovom vozraste v Voronezhskoi oblasti.

OBJECTIVE: To study the clinical course, the rate of progression and the results of the disease modifying treatment (DMT) in pediatric-onset multiple sclerosis (POMS) patients in the Voronezh region. MATERIAL AND METHODS: The clinical characteristics of the course of relapsing-remitting multiple sclerosis (MS) were analyzed among 51 POMS patients and 51 patients with the adult-onset MS (AOMS). The clinical course was assessed based on the Expanded Disability Status Scale (EDSS) score, the average annual frequency of exacerbations and the rate of disease progression before and during DMT. RESULTS: There were no statistically significant differences in EDSS scores 11 years after the onset of MS between the groups: POMS patients achieved moderate disability by the age of 25, and AOMS patients showed stabilization of their condition during DMT by the age of 36. During changing first-line drugs in patients with POMS, there was again an increase in disability with the transition of 25% of patients to the group with secondary progression of MS on average after 8 years of DMT. CONCLUSION: The onset of the disease in children and adolescents leads to a significant decrease in their quality of life and the deterioration of all activities throughout their lives. To prevent the progression of the disease it seems appropriate to transfer patients with POMS immediately to second-line DMT when signs of a suboptimal response to first line DMT appear, and in the case of a rapidly progressive course to start therapy with second-line drugs.

[Medical-biological approach to development of feeding ration for sportswomen--members of Russia weight lifting team].

Physical status of 12 sportswomen - member of Russia weight lifting team during training period was analiysed. Anthropometric, bioimpedance methods with analyzer ABC-01 "Medass" in observation [body weight Index (index Ketle-2), waist circle/hip circle index] and body compositions were used (adipose, bone and muscle tissues masse). Data obtained when bioimpedance method was used indicated: normal body weight and relative adipose mass to body weight - in 92%, surplus - in 8% of cases; body weight Index and normal level of adipose mass - in 75%, surplus - in 25%; active cellular mass and bone mass and general body liquid were found to be normal. 14-day feeding ration for sportswomen were analysed and 3 times per day food intakes was found to be not optimal. Data obtained let to recommend set of daily food products to compose optimal ration for present sportswomen group when energetic value of 4500 kKal and high nutritional value would be provided.

Clinical and Genetic Characteristics of Preeclampsia.

Preeclampsia (PE) is a severe complication of pregnancy accompanied by arterial hypertension, edema, or proteinuria with impaired functioning of various organs and systems. It is also an important medical and social problem, which has been one of the leading causes of maternal and perinatal mortality and morbidity worldwide. Despite the achievements of modern medicine, the etiology of this pathology is still unknown. Recently, many scientists have especially focused on the study of genetic factors underlying the etiopathogenesis of PE, namely, the contribution of individual polymorphic loci of various candidate genes. The current study aimed to investigate the clinical characteristics of PE and the contribution of the polymorphic loci rs1042838 of Progesterone Receptor (PGR) gene and rs8068318 of the T-Box Transcription Factor 2 (TBX2) gene to the development of PE. The study was conducted on 219 women with PE with the mean±SD age of 26.52±5.51 years and 329 women with the physiological course of pregnancy as the control group with the mean±SD age of 26.27±4.88 years. In total, 64.20%, 68.29%, 16.44%, 98.63%, and 35.48% of women with PE had increased systolic and normal diastolic blood pressure (SBP and DBP) values, proteinuria, edema, and overweight (BMI≥25), respectively. In the control group, 100%, 1.53%, 1.12%, and 35.48% of cases had normal SBP values with no proteinuria, DBP>90 mm Hg, edema, and overweight (BMI≥25), respectively. An association was observed between the CC genotype of the rs8068318 polymorphism of the TBX2 gene with the risk of developing PE in women with PE (OR=2.12, 95%CI: 1.14-3.92, P=0.02). In addition, there was an association between the rs8068318 TBX2 polymorphic locus with lower SBP (Me=140, Q25 - Q75 130 - 142.5, P=0.01) and PBP (Me=50, Q25 - Q75 40 - 55, P<0.01). According to the GeneCards database, the TBX2 gene, a member of a phylogenetically conserved gene family, is located on the long arm of chromosome 17 and encodes the TBX2 T-box transcription factor protein, which is a regulator of the transcriptional activity of various genes (i.e., it suppresses the expression of CDKN2A (p19/ARF), inhibits cyclin-dependent kinase p21 Cip1 (CDKN1A), and affects the expression of MYC, RAS, BRCA1, and BRCA2 genes).

[Actual nutrition and physical state of athletes of the national sleigh team of Russia].

An assessment of actual nutrition and physical fitness of athletes of the Russian national sleigh team are presented, and the need to develop for them the individual well-balanced diet that will reimburse the amount of consumed energy and nutrients and help athletes to obtain high results.

[Attractive and immunosuppressive properties of volatile secretions induced in mice separately and combine influence of ionizing radiation and cyclophosphamide].

We have studied the properties of volatile secretions induced in laboratory mice, separately or combined influence of ionizing radiation and cyclophosphamide. The reaction of preference-avoidance response in the T-maze revealed that the urine samples of mice in the early period after exposure to ionizing radiation in doses of 2 and 4 Gy contain attractive for intact animals volatile components. While the introduction of cyclophosphamide in doses of 20 or 35 mg/g provokes the secretion of volatile components of aversive, repealing properties towards of intact individuals. Combined exposure to these factors leads to a reciprocal compensation of the impact of volatile secretions in the preference-avoidance response of intact mice. The examined effects depend to some extent on the dose of radiation and cyclophosphamide. Induced both by separate and associated exposure to radiation and cyclophosphamide volatile allocation have depressing effect of the humoral immune response, regardless of whether they are attractive or aversive properties. We are discussing their role in the mechanisms of chemosignaling.

Effect of dopamine on viability of BHK-21 cells.

We studied the effects of dopamine added to culture medium on survival of floating or adherent BHK-21 cells differing by organization of actin cytoskeleton. The viability of floating cells more drastically decreased with increasing dopamine concentration and duration of exposure than that of adherent cells. The cells worse adhered to the substrate and formed a monolayer. The formed monolayer degrades, cell borders become blurred, cells, polygonal in the control, are rounded. Preliminary blockade of dopamine receptors with haloperidol, inessential for cell survival and morphology, does not prevent the destructive effect of dopamine on the cells. Ultrastructural study revealed increased density of filamentous actin threads in deep compartments of cell cytoplasm after dopamine treatment, this increase being more pronounced in cells grown in suspension. Bearing in mind the polymerizing effect of dopamine on globular actin in vitro and the fact that the content of this protein in floating cells is higher than in adherent cells, we can conclude that the decrease in viability of BHK-21 cells is caused by interaction of dopamine with cytoplasmic globular actin.

[Effect of hypoxenum on bioenergetic processes in mitochondria and the activity of ATP-sensitive potassium channel].

The effect of hypoxenum on bioenergetic processes in heart and liver mitochondria of rats, connected with respiration, the generation of hydrogen peroxide, and the activity of ATP-sensitive K-channel ((mitoK)ATP) has been studied. It was shown that hypoxenum in the concentration range of 0.05-10 microg/ml stimulates respiration, increases the coupling in the respiratory chain, and enhances the formation of H2O2 and energy-dependent swelling associated with potassium transport in mitochondria. Hypoxenum removes the inhibitory effect of ATP on the energy-dependent swelling of mitochondria and partially reduces the accumulation of H2O2 in the presence of ATP. The role of antihypoxic and antioxidant action of hypoxenum associated with the activation of (mitoK)ATP is discussed.

[The drug hypoxen--a new inhibitor of mitochondrial respiration and mitochondrial dehydrogenases].

The effect of hypoxen on the oxygen consumption and activity of dehydrogenases in rat liver mitochondria has been studied. The addition of hypoxen to mitochondria caused a speed reduction of phosphorylating and uncoupling respiration. The minimal effective concentration of hypoxen was 15 microg/ml with succinate, 60 microg/ml with pyruvate or palmitoylcarnitine, and 120 microg/ml with glutamate as the substrates. The activities of malate, glutamate, and succinate dehydrogenases in mitochondria were significantly decreased by the effect of hypoxen.

The Oral Mucosa Status and the Correlation between the Functional Parameters and the Level of Nitric Oxide Metabolites in Saliva among Patients with GERD.

The study involved 91 patients (48 women and 43 men), aged from 18 to 70 years with GERD. All patients underwent the clinical dental examination according to a single scheme including general clinical manifestations (nausea, single vomiting, belching, heartburn, pain in the epigastrium and around the navel, and poor appetite) and dental manifestations of GERD. The objective assessment of the dental status of the examined patients included the measurement of the functional parameters of the mixed saliva, buffer capacity (BC) of saliva, and the detection of the nitric oxide metabolites (NOx) content in saliva from the right parotid salivary gland ("SRPSG") and in blood serum using the indirect method based on the determination of the stable metabolites: nitrates and nitrites using the Griess reaction. It was established that salivation rate among patients with GERD with the prevailing of ACR and SACR was at the lower limit of normal values (0.32 + 0.19 ml/min), and the salivation rate among patients with the prevailing of SALCR was low (0.10 + 0.04 ml/min). The BC of saliva among patients with the prevailing of ACR and SACR was high (9.07 + 1.23 mmol eq/l and 9.40 + 1.71 mmol eq/l, respectively) and was reduced among patients with the prevailing of SALCR (7.63 + 0.18 mmol eq/l). The NOx level in SRPSG among patients with GERD was increased (especially in Group 3 (20.93 + 11.23 umol/l)). The direct correlation between the indicators of sialometry, the level of the BC of saliva, and the NOx level in SRPSG were established during the study.

Heterosynaptic potentiation of cholinergic excitatory postsynaptic responses of command neurons in the common snail.

Short-term heterosynaptic potentiation of cholinergic excitatory postsynaptic currents and potentials in defensive behavior command neurons was found to be evoked by stimulation of the visceral sac in the common snail. It is suggested that a mechanism increasing the choline resistance of the postsynaptic zones of command neuron membranes is involved in potentiating the excitatory postsynaptic responses to sensory stimulation.