Evaluation of Collagenase Activity from Crab Hepatopancreas in Different Model Systems.

The effect of Kamchatka crab hepatopancreas containing three collagenolytic isoenzymes Collagenase KK and proteinases of Streptomyces lavendulae on metabolic activity and cell death were carried out on in vitro models. It was shown that changes in the protein structure under the influence of Collagenase KK occurred earlier than under the effect of bacterial proteinases. At the same time, activity of Collagenase KK was significantly higher than that of bacterial proteinases (p<0.01). Both preparations had a pronounced time- and dose-dependent effects on metabolic activity of cells. Collagenase KK had low cytotoxic effect, and cells mainly died by apoptosis. Thus, hepatopancreas collagenase has a high activity and proapoptotic effect on cells and can be used in low concentrations for enzymatic disaggregation of tissues.

[The effect of overweight on the dental status and biochemical parameters of the oral fluid]. / Vliyanie izbytochnoi massy tela na stomatologicheskii status i biokhimicheskie parametry rotovoi zhidkosti.

OBJECTIVE: To increase the effectiveness of the diagnosis and treatment of dental diseases, based on the study of anthropometric and bioimpedance analysis data in adolescents and adult patients to identify the effect of overweight on dental status. MATERIAL AND METHODS: The study involved 60 adolescents aged 15-18 years, 28 of whom were overweight, and 32 with normal body weight. All 52 adult patients aged 30-50 years who participated in the study were overweight (body mass index >25 kg/m2) and had chronic generalized periodontitis. In all patients dental status was assessed by DMF and PMA index, Silness-Loe and Stallard hygienic indices, Muleman bleeding index, Green-Vermillion tartar index. Biochemical parameters of the oral fluid were also evaluated: malondialdehyde, elastase, urease, catalase and lysozyme activity. The adolescents underwent an anthropometric study with the determination of body mass index. Adult patients underwent bioimpedance analysis of body composition to determine the main indicators of fat metabolism in the body: body mass index, fat mass (kg), the proportion of adipose tissue (%) and the mass of extracellular fluid (kg). RESULTS: The study showed that overweight in patients of different ages worsens their dental status and biochemical parameters of oral fluid. CONCLUSION: The inclusion of an anthropometric study with the determination of body mass index and bioimpedance analysis of body composition in the examination of dental patients will allow the development of individual programs for the prevention of dental diseases, implementing a personalized approach to the provision of medical and preventive care.

[The impact of primary prevention programs of dental diseases on the main trends in the development of dental services]. / Vliyanie programm pervichnoi profilaktiki stomatologicheskikh zabolevanii na osnovnye tendentsii razvitiya stomatologicheskoi sluzhby.

The vision of the global strategy on oral health is universal health coverage for oral health for all individuals and communities by 2030, enabling them to enjoy the highest attainable state of oral health and contributing to healthy and productive lives (WHO, 2022). To achieve this goal, it is necessary to ensure the development of the dental care system in Russia on the basis of primary prevention of dental diseases. OBJECTIVE: Analysis of the methodology for the development, implementation and evaluation of programs for the primary prevention of dental diseases and their impact on the main trends in the development of dental services. MATERIAL AND METHODS: The main research methods were search for publications, analysis and systematization of information on the methodology for developing, implementing and evaluating programs for the primary prevention of dental diseases. RESULTS: Despite the single main goal of dental disease prevention programs, the analysis of the methodology of their construction and implementation must be carried out considering their impact on the main trends in the development of dental services. CONCLUSIONS: The main directions of development of the methodology for the development, implementation, and evaluation of primary prevention programs of dental diseases should include the use of indicators of oral health recognized by the international community, which allow us to trace the degree of their influence on the development of the system of dental care.

[Monitoring the implementation of the compulsory health insurance program for the prevention of dental diseases]. / Monitoring realizatsii programmy obyazatel'nogo meditsinskogo strakhovaniya po profilaktike stomatologicheskikh zabolevanii.

The purpose of the study is to monitor the implementation of the CHI program for the prevention of dental diseases. Based on the analysis of the monitoring results, the need for adjustment and direction of improvement of the observed process was identified. During the period from 2013 to 2018, there was a slight increase in the volume of preventive care per 1 insured citizen, from 0.186 visits to 0.211, which does not meet the recommended indicator and is insufficient. In the dynamics of expenses per 1 insured citizen, there is a trend of significant growth of 1.93 times. The cost of 1 visit with a preventive purpose in the dynamics also increased by 1.71 times, amounting to 603.16 rubles in 2018. it is Necessary to specify in detail the methodology for conducting a preventive examination, its interpretation and the list of services provided during this period.

[Glycol and Phosphate Depot Forms of 4- and/or 5-Modified Nucleosides Exhibiting Antibacterial Activity].

Resistance developed to the majority of drugs used to treat infectious diseases warrants the design of new compounds effective against drug-resistant strains of pathogens. Recently, several groups of modified nucleosides have been synthesized and showed significant antibacterial activity in vitro, but their further studies were difficult to undertake because of their low solubility in aqueous solutions. Nevertheless, new compounds, well soluble in water-organic solutions, were synthesized and found to be more effective in inhibiting the growth of Gram-positive bacteria and mycobacteria. The water-soluble forms of modified nucleosides under study were assumed to be their depot forms. To check the assumption, the compounds were tested for hydrolysis in various media and their molecular docking was performed into the active center of the putative target, Mycobacterium tuberculosis flavin-dependent thymidylate synthase ThyX. Computer modelling showed that the water-soluble analogs do not act as ThyX inhibitors, supporting the assumption of their depot nature. The compounds were resistant to chemical hydrolysis but were hydrolyzed when incubated with porcine liver carboxylesterase, human serum, or Staphylococcus aureus 209P. The results demonstrate that the compounds are most likely depot forms of modified nucleosides.

[The dinamics of preventive dental visits in mandatory health insurance system]. / Dinamika poseshchenii s profilakticheskoi tsel'iu v sisteme obiazatel'nogo meditsinskogo strakhovaniia po profiliu 'Stomatologiia'.

The analysis on the current standards of the amount of Medical care provided for dental diseases treatment in Russia within mandatory health insurance program has been made. The study revealed more than 80-fold difference in the number of visits for one insured patient in different regions of Russian Federation. The annual increase in financial expenses covering dental visits for prophylactic purposes is identified. It is recommended that the Federal Fund of Mandatory Health Insurance to state a clear definition to the term 'Dental visit for prophylactic purposes', while the territorial funds should strengthen the requirements for approved reporting forms submition.

First detection of VIM-4 metallo-ß-lactamase-producing Escherichia coli in Russia.

An Escherichia coli isolate co-producing VIM-4 metallo-ß-lactamase and CTX-M-15 extended spectrum ß-lactamase was recovered from the urine of a patient with head trauma in Moscow, Russia. The bla(VIM-4) and bla(CTX-M-15) genes were carried, respectively, by transmissible plasmids of IncW and IncI1 groups. The nucleotide sequence of the VIM-4-encoding integron was nearly identical to that of In416, which represent a large group of structurally related integrons previously found in Enterobacteriaceae all around the Mediterranean basin. This is the first report of a metallo-ß-lactamase-producing E. coli in Russia.

[The course of virus infection of Lycopersicon esculentum plants at high concentrations of heavy metals]. / Perebih virusnoï infektsiï u roslyn Lycopersicon esculentum pri vysoykh kontsentratsiakh vazhkykh metaliv.

Effect of certain compositions of heavy metals on the course of the system infection of TMV in plants of tomatoes Lycopersicon esculentum L. under field conditions was investigated. It is shown that the simultaneous influence of the virus infection and heavy metals in concentration of 10 MPC (maximum permissible concentration) resulted in the decrease of the content of total chlorophyll in the investigated plants by degradation of chlorophyll A; that was more considerable decrease as compared with separate influence of each of the factors. Treatment of plants by toxical concentrations of lead and zinc led to inconsiderable delay in development of the system TMV infection together with the increase of the virus content in plants 2 and more times, that evidences for the probable synergism in the action of the given heavy metals and virus infection. On the contrary, copper, though it decreased the content of total chlorophyll, manifested the protective properties and restricted essentially the virus accumulation in a plant as a whole.

[Changes in the level of DNA minor bases in salivary gland inflammation and its correction with antioxidants]. / Izmenenie soderzhaniia minornykh osnovanii DNK pri vospalenii sliunnoi zhelezy i ego korrektsiia antioksidantami.

In the experiment with guinea pigs at contact sialoadenitis reproduction (karragenine inflammation of soft tissues of the oral cavity) it was established that in DNA of salivary glands cells and the neighbouring tissues the content of 5-methylcytosine was decreased, it being normalized at "Triovit" antioxidant complex together with cvercitynum correction. The content of DNA 8-hydroxyguanine is characterized by a wide range of data and in absolute values increases in inflammation, but decreases after antioxidant correction. The role of the changes discovered is discussed.

[The sources of active forms of oxygen in the normal oral tissues and in pathology]. / Istochniki aktivnykh form kisloroda v tkaniakh rotovoi polosti v norme i pri patologii.

In the experiment on rats the changes of the superoxydanionradical production from microsomal and mitochondrial oxidation and tissue phagocytes of the saliva gland and parodontium in contact sialadenitis, spontaneous parodontitis, after ionizing radiation have been investigated. It is supposed that an increase or decrease of superoxydanionradical production from different sources demands a differentiated corrective approach.

Membrane topology and cellular location of the Treponema pallidum glycerophosphodiester phosphodiesterase (GlpQ) ortholog.

Recent reports that isolated Treponema pallidum outer membranes contain an ortholog for glycerophosphodiester phosphodiesterase (GlpQ) (D. V. Shevchenko, D. R. Akins, E. J. Robinson, M. Li, O. V. Shevchenko, and J. D. Radolf, Infect. Immun. 65:4179-4189, 1997) and that this protein is a potential opsonic target for T. pallidum (C. E. Stebeck, J. M. Shaffer, T. W. Arroll, S. A. Lukehart, and W. C. Van Voorhis, FEMS Microbiol. Lett. 154:303-310, 1997) prompted a more detailed investigation of its physicochemical properties and cellular location. [14C]palmitate radiolabeling studies of a GlpQ-alkaline phosphatase fusion expressed in Escherichia coli confirmed the prediction from DNA sequencing that the protein is lipid modified. Studies using Triton X-114 phase partitioning revealed that the protein's amphiphilicity is due to lipid modification and that a substantial portion of the polypeptide is associated with the T. pallidum peptidoglycan sacculus. Three different approaches, i.e., (i) proteinase K treatment of intact treponemes, (ii) indirect immunofluorescence analysis of treponemes encapsulated in agarose beads, and (iii) opsonophagocytosis of treponemes incubated with antiserum against recombinant GlpQ by rabbit peritoneal macrophages, confirmed that GlpQ is entirely subsurface in T. pallidum. Moreover, rabbits hyperimmunized with GlpQ were not protected against intradermal challenge with virulent treponemes. Circular dichroism spectroscopy confirmed that the recombinant form of the polypeptide lacked discernible evidence of denaturation. Finally, GlpQ was not radiolabeled when T. pallidum outer membranes were incubated with 3-(trifluoromethyl)-3-(m-[125I]iodophenyl)-diazarene, a photoactivatable, lipophilic probe which promiscuously labels both proteins and lipids within phospholipid bilayers. Taken as a whole, these studies indicate that the T. pallidum GlpQ ortholog is a periplasmic protein associated predominantly with the spirochete's peptidoglycan-cytoplasmic membrane complex.

Identification of homologs for thioredoxin, peptidyl prolyl cis-trans isomerase, and glycerophosphodiester phosphodiesterase in outer membrane fractions from Treponema pallidum, the syphilis spirochete.

In this study, we characterized candidate rare outer membrane (OM) proteins with apparent molecular masses of 19, 27, 38, and 38.5 kDa, which had been identified previously in OM fractions from Treponema pallidum (J. D. Radolf et al., Infect. Immun. 63:4244-4252, 1995). Using N-terminal and internal amino acid sequences, a probe for the 19-kDa candidate was PCR amplified and used to screen a T. pallidum genomic library in Lambda Zap II. The corresponding gene (tlp) encoded a homolog for periplasmic thioredoxin-like proteins (Tlp), which reduce c-type cytochromes. A degenerate oligonucleotide derived from the N terminus of the 27-kDa protein was used to PCR amplify a duplex probe from a T. pallidum genomic library in pBluescript II SK+. With this probe, the corresponding gene (ppiB) was identified and found to code for a presumptive periplasmic cyclophilin B-type peptidyl prolyl cis-trans isomerase (PpiB). We postulate that PpiB assists the folding of proteins within the T. pallidum periplasmic space. The N terminus of the 38-kDa candidate was blocked to Edman degradation. However, internal sequence data revealed that it was basic membrane protein (Bmp), a previously characterized, signal peptidase I-processed protein. Triton X-114 phase partitioning revealed that despite its name, Bmp is hydrophilic and therefore likely to be periplasmic. The final candidate was also blocked to Edman degradation; as before, a duplex probe was PCR amplified with degenerate primers derived from internal sequences. The corresponding gene (glpQ) coded for a presumptively lipid-modified homolog of glycerophosphodiester phosphodiesterase (GlpQ). Based upon findings with other treponemal lipoproteins, the hydrophilic GlpQ polypeptide is thought to be anchored by N-terminal lipids to the periplasmic leaflet(s) of the cytoplasmic membrane and/or OM. The discovery of T. pallidum periplasmic proteins with potentially defined functions provides fresh insights into a poorly understood aspect of treponemal physiology. At the same time, however, these findings also raise important issues regarding the use of OM preparations for identifying rare OM proteins of T. pallidum.