[Evaluation of the dynamics of prosthetic dental care provided to patients with the complete absence of teeth in the Perm district]. / Otsenka dinamiki ob"ema ortopedicheskoi pomoshchi patsientam s polnym otsutstviem zubov v Permskom krae.

BACKGROUND: Currently, one third of the world's population aged 65-74 years has partially or completely missing teeth. In our country, the data of epidemiological studies are scattered. OBJECTIVE: The aim of the study is assessment of the dynamics of the volume of orthopedic care provided to patients with complete absence of teeth in the Perm Region. MATERIAL AND METHODS: The study of the volume of orthopedic care provided to patients with complete absence of teeth was carried out on the basis of a detailed retrospective analysis of their referral to dental clinics in the Perm Region for the period from 2014 to 2020. To assess the long-term dynamics of indicators of the volume of orthopedic care provided to patients with complete absence of teeth, the least squares method was used-gradation of growth rates. RESULTS: This investigation shows the trend of increasing the volume of orthopedic care provided to patients with complete absence of teeth for the period from 2014 to 2020. The average annual growth rate was 5.2%, this situation may be partly due to the accumulation of chronic inflammatory diseases of the oral cavity in the population. Among the patients who applied for dental orthopedic care with complete absence of teeth, women prevailed - 61.8%. The average age of patients with complete absence of teeth was 63.2±1.6 years. CONCLUSION: The trend of increasing the volume of orthopedic care provided to patients with complete absence of teeth in the Perm Region indicates unfavorable changes in the overall dental morbidity over the period 2014-2020 and increased availability of removable dentures with complete loss of teeth. Forecasting the volume of care for patients in this category is important for planning preventive measures.

Express detection of Escherichia coli and bacteria of the Escherichia coli group at the oral cavity.

Currently, in the diagnosis of diseases, a decisive place is given to laboratory methods, which should be informative, relatively simple to perform and rapid. The article describes the approbation of a method for rapid detection of Escherichia coli and bacteria of Escherichia coli group in the oral cavity. Research involved 44 volunteers, who were sampled from the oral cavity, followed by incubation in Koda's medium. The study used oral (n=11) and gingival fluids (n=11); smears-prints from the oral mucosa (n=11); dental biofilm (n=11). After 24 hours, the change in color and transparency of the medium was assessed. The preservation of the initial green color and transparency by the medium meant the absence of E. coli and bacteria of Escherichia coli group in the sample. A change in the color of the medium to yellow, turbidity and / or the formation of bubbles indicated the presence of E. coli and bacteria of Escherichia coli group. In parallel, the material was inoculated onto Endo agar, followed by identification of strains to species. As a result of the study, a complete coincidence of the results of the classical bacteriological method and the method using Koda medium was shown. In the latter case, a significant advantage is the speed of obtaining the result (18-20 hours), in contrast to the classical method, the interpretation of the results of which is available only after 72 hours or more. All of this is in line with the state of the art in clinical microbiology and rapid diagnosis based on «point-of-care testing / doctor's office¼ diagnostic principle. The presented method can be successfully applied in clinical practice for topical diagnosis of microorganisms E. coli and bacteria of Escherichia coli group in the oral cavity.

[Colonization activity of conditionally pathogenic microorganisms and features of biofilm formation on the surface of thermoplastic polymer material]. / Kolonizatsionnaya aktivnost' uslovno-patogennykh mikroorganizmov i osobennosti obrazovaniya bioplenki na poverkhnosti stomatologicheskogo termoplastichnogo polimernogo materiala.

THE PURPOSE OF THE STUDY: Was to assess the microbial colonization and biofilm-forming activity of conditionally pathogenic microorganisms in vitro to samples of acrylic-free thermoplastic polymer material and to assess the possibility of its use for the manufacture of removable tooth splinting structures by injection method. MATERIALS AND METHODS: The reference strains S. aureus, S. epidermidis, S. pyogenes, E. faecalis, E. coli, and C. albicans. Biofilm-forming activity was assessed by the level of ethanol extraction of a 0.1% aqueous solution of gentian violet by measuring on a PowerWave X microplate reader (USA). RESULTS: A low degree of severity of microbial colonization and sufficient colonization resistance to the formation of a microbial biofilm of E. coli (0.374±0.056 cu opt. Pl.), S. aureus (0.272±0.039 cu. Opt. Pl.), S. epidermdis (0.299±0.028 cu opt. Pl.), S. ryogenes (0.399±0.069 cu opt. Pl.), E. faecalis (0.401±0.089 cu opt. Pl.). Moreover, C. albicans strains form a more pronounced biofilm (0.425±0.104 cu opt. Pl.) in comparison with other strains studied in the experiment. CONCLUSION: Samples from a thermoplastic polymer based on polyoxymethylene have satisfactory biomedical characteristics and can be used at the stages of dental orthopedic treatment of patients without oral candidiasis.

A new approach for the study of the morphological structure of microbal biofilms.

Microbial biofilms are heterogeneous, moving and constantly changing communities of microorganisms, often of various taxons. Approaches to study and assessment anti-biofilm drugs widely available today do not adequately assess their effects, while the results of studying the interaction of drugs with components of the film composition can provide them the right choice. The aim of investigation was to test a new method of morphological evaluation of biofilms. To form biofilms, we used an approach when the slide was placed at an angle of 30o-45o relatively to the Petri dish, and a suspension of test strains S. epidermidis in peptone broth was poured into the space between the Petri dish and the slide. A sterile cotton swab moistened with distilled water was placed next to the glass slide to create optimal humidity. The system was placed in a thermostat for 24 hours. The formed films were examined under a microscope using the DCM 310 video eyepiece and the Scope photo x86,3.1.312 program that allowed to conduct a complete morphometric study of the film: select layers, channels, cavities and make measurements, and then save the results on electronic media in jpg file format. Microscopy of the stained slides revealed that the biofilm has a layered structure. In each image obtained using a video eyepiece, it was possible to differentiate 4 layers. From the border of the two media to the inside: the fragmentation layer, the dense layer, the matrix substance layer, and the last one - the persistence layer. Channels of different diameters (from 10 to 24 microns) are observed across the entire thickness of the biofilm. Thus, used approach allows us to visualize and evaluate the structure of microbial biofilm, measure the thickness of layers and channel diameters. In addition, this method can be used to study the effect of antimicrobial drugs on bacterial films.

[Etiology of purulent-inflammatory processes in the genital tract: suspicions of clinicians and problems of laboratory confirmation.]

Despite the long history of the study, laboratory diagnosis of gonococcal infection remains a complex task that does not have a clearly regulated effective solution. Aim of investigation was to assess the species diversity of the microbiota of the genital tract of men and women with suspected acute genital gonococcal infection (AGGI) using test systems of Russian manufacturers. A study of samples of the contents of the urethra of 69 men and posterior vaginal fornix fluids of 33 women of reproductive age with characteristic clinical manifestations and a presumptive diagnosis of AGGI was made. Cultivation was carried out using elective culture media with subsequent identification of strains by biochemical properties. Detection of DNA of Neisseria gonorrhoeae, Staphylococcus spp., Streptococcus spp., Enterobacteriaceae, Gardnerella vaginalis, Atopobium vaginae, Lactobacillus spp. performed by PCR using Vektor-Best and InterLab Service kits (Russia). All patients were divided into groups according to the results of the bacteriological method and PCR. A metagenomic study of 16S ribosomal RNA samples was performed on the Illumina MiSeq platform using the MiSeq Reagent Kits v3 kit (600-Cycle Kit). Statistical analysis of the data was performed using criterion x2. As a result of a laboratory study, the presumptive clinical diagnosis of «AGGI¼ found its bacteriological confirmation in 35.3% of cases only, among which fragments of the N. gonorrhoeae genome were detected in 63.9% of the samples only. Moreover, a wide variety of microorganisms in the genital tract of both men and women was found in metagenomic analysis. However, this technique does not allow us to assess the viability of the detected bacteria, and the microflora spectrum is excessively wide. In addition, the high level of genetic polymorphism of different strains of N. gonorrhoeae complicates the interpretation of the results. Deciphering the composition of microbiota allows the use of InterLab Service kits. The decoding of the etiology of purulent-inflammatory processes in the genital tract, which presents serious difficulties, is greatly facilitated by the use of Russian kits for molecular genetic analysis, which, in our opinion, provide the necessary and sufficient information for practice.

[The determination of biofilm composition of gram-positive bacteria.]

Current methods of biofilm imaging do not support a differentiated assessment of its composition, since it is not possible to establish a substrate stained with crystal violet, as this dye can form complexes with both intracellular and extracellular structures. This approach does not adequately assess the anti-biofilm effects of drugs, while the results of studying the interaction of drugs with biofilm components can ensure their most correct choice. The aim of investigation was to study the possibility of applying the original modification of the current method to determine the ratio of the cellular part and the matrix of biofilms of gram-positive microorganisms. The biofilm components were analyzed using a two-step approach, when prepared biofilms of gram-positive microorganisms were stained with crystal violet for 5 minutes, followed by fixing the dye in bacterial cells with iodine solution, and then the colored products were dissolved with 95% alcohol: matrix components for 1 minute, total biofilm for 15 minutes, after which the composition of biofilms was estimated by the formula: M=(OP1/OP15)×100, Kb=100-M, where M is the proportion of the matrix,%; Kb - the proportion of the cellular component,%; OP1 - optical density of samples, when alcohol was allowed to dissolve the colored product for no more than 1 minute; OP15 - was the optical density of samples, when alcohol is allowed to dissolve the colored product for 15 minutes. It was shown that in the composition of the biofilm formed by the collection strain, the proportion of the matrix was 13.2%, and the cellular component accounted for 86.8%. When the same strain cultivated in the presence of an antibiotic, an increase in the biofilm matrix was observed, which is probably due to the compensatory response of the microorganism to the action of the antibiotic. The proposed approach to the study of biofilms makes it possible to evaluate its component composition. Obtaining additional information in this way can provide, inter alia, an increase in the effectiveness of antimicrobial therapy while reducing the study time.

[Characteristics of lactobacteria strains, having diagnostic significance in gynecological practice.]

BACKGROUND: Nowadays, in the presence of wide diagnostic possibilities, laboratory diagnostics of microecological disorders of vaginal biotope are often limited to clinical data, microscopic examination results and the use of a culture method. However, with such a complex it is impossible to get an answer about the functional activity of microorganisms. The aim of investigation was to evaluate the information content of a combined study of growth parameters and the ability to produce lactic acid by clinical strains of Lactobacillus spp. to characterize the state of the microecology of the cervical-vaginal biotope. MATERIALS AND METHODS: Studied the growth kinetics of strains of lactobacilli isolated from the detachable posterior vaginal fornix. The concentration of lactic acid in the medium was determined using the "Lactic Acid - Olvex" kit (Russia). Samples were taken every 12 hours of cultivation. Statistical analysis of the results was performed using the methods of descriptive statistics, Student's t-test. RESULTS: It has been shown that 44% of Lactobacillus spp. to 72 hours of cultivation did not reach the phase of exponential growth. The remaining strains from 12-24 hours passed into the phase of exponential growth. In the production of lactic acid, the strains that are in the lag-phase did not differ from each other, since they practically did not synthesize this metabolite. Among the remaining strains that are in the phase of exponential growth, some did not produce lactic acid, others increased their lactate production every hour. Part of the strains reached the peak of acid production by 36 hours and by 72 hours some decrease in synthetic activity was observed. CONCLUSIONS: It has been shown that for most vaginal strains of Lactobacillus spp. characteristic variability of the duration of the adaptation period and the level of functional activity. In addition, only a small number of clinical strains produced lactic acid 24 hours after the start of cultivation. Therefore, to adequately assess the microecological status of the vaginal biotope, microscopic examination of both native material and cultures isolated on special nutrient media is not enough. It seems that, along with the use of modern methods of genetic analysis, the determination of in vitro growth characteristics, primarily lag-phase duration, and lactate production by lactic acid bacteria strains can clarify many issues related to the formation of dysbiotic states, in particular, in the vaginal biotope, and will also serve to increase the effectiveness of the prescribed treatment.

[Determination of biofilm forming activity of microorganisms on synthetic polymeric materials.]

Microorganisms are able to form biofilms on surfaces of biotic and abiotic nature. In turn, in human biotopes there are optimal conditions for the implementation of biofilm-forming activity. Moreover, in medical practice, polymeric materials are often used for drainage or prosthetics, which can also be successfully colonized by bacteria. However, in laboratory practice, the formation of biofilms is usually evaluated on glass or polystyrene. The purpose of the study is to evaluate the methodological features of studying the biofilm-forming activity of microorganisms on the surface of synthetic polymeric materials. We used strains of Staphylococcus aureus ATCC 25923, Escherichia coli K-12, Candida albicans ATCC 10231, as well as synthetic polymeric materials - DentLight Flow light-curing composite material (nano-hybrid fluid composite; Russia), glass ionomer chemical curing Fuji 1 (Japan), cement for temporary fixation of orthopedic constructions TempBond NE (USA), acrylic, polyurethane and polyvinyl chloride. The formation of biofilms in flat-bottomed ELISA plates in this study was considered as a control group. If the polymer belonged to cold curing materials, sterile flat-bottomed tablets were used, the bottom of which was filled with a thin layer of plastic. After hardening of the plastic, biofilms were formed in the tablets. In the second series of experiments, hot cured materials cut into equal parts 5×5×1 mm in size were placed in the wells of a plate and again used to determine biofilm formation with subsequent coloring. To extract the dye, the pieces were transferred to a new plate to exclude the amount of film biomass formed on the walls of the plate wells. In both cases, cultivation was carried out at 37° C for 24-48 hours. The biomass of the film was stained with fuchsin. Statistical data processing was performed using t-Student criterion. For the threshold level of significance, the value p <0.05 was taken. It is established that the proposed options for determining biofilm forming ability are available and indicative. It was revealed that the same microorganisms have individual biofilm formation indicators for each polymer material. The light curing dental composite and polyvinyl chloride exhibit the more pronounced antiadhesive properties than cements and polyurethane. Up to date, most of the studies of biofilm formation have been carried out using glass or polystyrene, which, as a rule, are not used for the manufacture of prostheses, catheters, drains, etc., which makes it difficult to assess the true film-forming activity of microorganisms. The proposed methodological approaches, especially the second option for preparing testing samples, solve this problem. In general, the proposed approaches to testing biofilm-forming activity on polymers are very simple to implement and generally available. For an adequate study of the biofilms formation, it will be advisable to use polymer materials, directly used in medicine, rather than polystyrene tablets, the material of which is found exclusively in laboratory practice.

[GUT AND VAGINAL MICROBIOTA IN FEMALE WITH REPRODUCTIVE DISORDERS ASSOCIATED WITH GASTROINTESTINAL DISTURBANCE].

The aim of research - a comparative study of the microflora of the vagina and bowel in women with secondary infertility and gastrointestinal distress. Materials and methods. We examined gut and vaginal microbiota in 17 infertile women according to the presence or absence of gastrointestinal diseases. We used a standard procedure for microbiological examination. Results. Certain changes of bacterial load and composition in both reservoirs were established. They allowed us to conclude the same trends in characterizing of the developing dysbiosis. Conclusion. Thus, the simultaneous studying of vaginal and gut microflora seems to be rational during the examination of women with childbearing disorders. It could possibly increase the treatment efficacy and/or prevent the development of some pathology.