Diabetic Nephropathy - possibilities of early laboratory diagnostics and course prediction (review of literature).

The medical and social significance of diabetes mellitus is determined by a progressive increase in the incidence of the disease, as well as the development of disabling and reducing the quality of life of macro- and microvascular complications in patients of working age, in particular, with the development of diabetic nephropathy (DN), which develops in one third of patients with type 1 and 2 diabetes. DN is the third leading cause of death after diseases of the cardiovascular system and oncological pathologies. In this regard, the identification of DN at the early stages is an important task both from the standpoint of prevention and a more favorable prognosis of the course of diabetes. This review presents data on the possibilities of early diagnosis of DN using blood and urine biomarkers, as well as information on their diagnostic and prognostic value. The analysis of the interpretation of the indicator of microalbuminuria in type 1 and 2 diabetes was carried out. In addition, the literature data of recent years on the informative value of determining markers of podocyte damage, on the role of oxidative stress products, immune-inflammatory factors, and vascular endothelial growth factor in the process of the onset and development of DN are presented. The possibility of their practical use for the diagnosis of DN in clinical practice is discussed. The literature search for this review was carried out using the databases of the RSCI, CyberLeninka, Scopus, Web of Science, MedLine, PubMed for the period from 2011 to 2021. using the following keywords: diabetes mellitus, diabetic nephropathy, markers of diabetic nephropathy; kidney damage, early diagnosis of diabetic nephropathy.

[Characterization of B1-cells during experimental leukomogenesis.]

BACKGROUND: Bovine leukemia causes a significant polyclonal expansion of CD5+, IgM+ B lymphocytes, known as persistent lymphocytosis (PL), in approximately 30% of infected cattle. However, it is not yet clear what happens to this subpopulation of B cells in the early period of infection of animals. PURPOSE: Quantitative characterization of IgM+ and CD5+ B cells during the immune response, which can provide important information on the mechanisms of lymphocyte priming in BLV infection. MATERIAL AND METHODS: The experiment used BLV-negative calves of black-motley breed at the age of 8 months (n = 11). Animals (n = 8) were intravenously injected with blood of a BLV-positive cow. Control calves (n = 3) were injected with saline. Studies were performed before and after infection on days 5, 7, 14, 21, 28 and 65 of the immune response. The determination of the number of B-lymphocytes in the blood was carried out by the method of immunoperoxidase staining based on monoclonal antibodies to IgM, CD5. RESULTS: As a result of the studies, it was found that the level of CD5+ B cells increases on the 14th day of the primary immune response, characterized by polyclonal proliferation of CD5+ B cells, which are the primary target for BLV. Our research data confirm that in the lymphocytes of experimentally infected cattle, surface aggregation of IgM and CD5 molecules on B-lymphocytes is absent. DISCUSSION: It is known that the wave-like nature of IgM synthesis, which was shown in previous studies, depends on a subpopulation of B1 cells. After 7 days of the immune response, IgM+ and CD5+ cells do not correlate, which shows their functional difference. The increase in CD5+ cells is probably not associated with B cells, but with T cells differentiating under the influence of the virus. CONCLUSIONS: A subset of B1 cells is the primary target of cattle leukemia virus. The 65th day of the immune response is characterized by the expansion of IgM+ B cells, a decrease in the number of CD5+ cells and a uniform distribution of receptors around the perimeter of the cells.

[Markers of endothelial dysfunction: pathogenetic role and diagnostic significance.]

Endothelial dysfunction (ED) is considered one of the pathogenetic mechanisms of a whole range of diseases. Detection of specific biochemical markers in the blood is an effective way to ED diagnostics that characterize the vascular endothelium state. This review highlights the pathogenetic role of the factors synthesized by endotheliocytes whose level changes in biological fluids reflect violations of the endothelium basic physiological properties: vasomotor function, thromboresistance, angiogenesis regulation, barrier and adhesion functions. In particular, the participation of nitric oxide metabolites, asymmetric dimethylarginine, endothelin-1, metabolic products of arachidonic acid, von Willebrand factor, thrombomodulin, vascular endothelial growth factor, vasohibine-1 and adhesion molecules in the onset and development of ED are reviewed. The diagnostic significances of factors damaging endothelium, such as C-reactive protein, homocysteine and 8-hydroxy-2'-deoxyguanosine, are discussed. In addition, the literature data of recent years about the prospects of clinical implication the detection of the above-mentioned factors which indicates structural and functional endothelial cells damage are given. Particular attention is paid to the ED markers detection prognostic significance and the possibility of their practical use for the ED diagnosis. The search of literature for the current review was conducted in RSIC, CyberLeninka, Scopus, Web of Science, MedLine and PubMed databases from 2012 to 2018 using the following keywords: endothelial dysfunction, nitric oxide, asymmetric dimethylarginine, endothelin-1, prostacyclin, thromboxane A2, epoxyeicosatrienoic acids, von Willebrand factor, thrombomodulin, vascular endothelial growth factor, vasohibin-1, adhesive molecules, C-reactive protein, homocysteine, and 8-hydroxy-2-deoxyguanosine.

[Detection of specific antibodies of classes G and M to bovine leukemia virus in the blood serum.]

INTRODUCTION: Bovine leukemia is a widespread infection worldwide, the causative agent of which is the bovine leukemia virus (BLV) in structural structure and functional features similar to human T-cell leukemia virus (HTLV-1 and HTLV-2) and It is considered as an actual medical and social problem. The study of the immune response in experimentally infected calves at an early stage of the disease development, synthesis of specific antibodies of classes G and M (IgG and IgM), diagnostic informativeness of detection of IgM in cattle leukemia is relevant and determines the purpose of this study. MATERIAL AND METHODS: Samples of blood and serum of cattle: animals experimentally infected with VLCRS, patients with cattle leukemia; control negative; specific to heterologous pathogens of cattle diseases. Indirect and sandwich variant enzyme-linked immunosorbent assay (ELISA); commercial ELISA kits (IDEXX, USA; Hema LLC, FKP Kursk Biofactory Firm BIOK, Russia) for the detection of specific IgG and IgM for BLV in the agar gel immunodiffusion reaction (RID). RESULTS: The humoral immune response develops shortly after infection - by 1-8 weeks. IgM are detected starting from the 3rd day, and IgG from the 7th day after infection. Up to 97% of coincidence of positive results in RID and indirect variant of TF ELISA based on monoclonal antibodies to cattle IgM (IgMbovine) were found. DISCUSSION: The dynamics of the synthesis of antibodies of classes M and G to the glycoprotein gp 51 BLV has a dosedependent wave-like character, is consistent with the levels of increase / decrease in the absolute and relative number of leukocytes / blood lymphocytes of infected calves. FINDINGS: Serum specific IgM was detected starting 3 days after infection with BLV. Early detection of IgM in serum of cattle can be used as an additional test for the detection of sick animals.

[MICROBIAL COMMUNITIES ON KIDNEY STONES].

The clinical material obtained surgically in patients with kidney stone disease (KSD) was tested for content of the stone microflora using PCR and standard microbiological methods. It was demonstrated that about 50% of stones in patients with KSD were infected with various infection agents as observed using standard microbiological and molecular genetic methods. The percentage of detection of the Mycoplasma hominis using cultural method is lower than the percentage detected using PCR, which is due to difficult isolation and cultivation, as well as DNA fragments of mycoplasma observed after antibiotic therapy. Studies based on modern microscopy methods showed that microorganisms on the surface of the kidney stone formed multispecies biofilms.

[Renal calculus microflora in urolithiasis and search for agents of control of biofilms formed by uropathogenic bacteria].

AIM: Study bacterial biofilms in native material (renal calculus) by electron microscopy method and developmeit of biofilm model by isolates in vitro on sterile calculi of various chemical composition. MATERIALS AND METHODS: Bacterial spectra of microflora of renal calculus lavages were studied, isolated pure cultures were identified up to species. Comparisons of urine microflora obtained before operation in patients with urolithiasis with microflora of removed renal calculi were carried out. RESULTS: Urease activity and genes coding pathogenicity factors were detected, and the ability to form biofilms by isolates was studied. Model of formation of biofilms in vitro on sterile renal calculi was developed and candidate agents reducing the biofilm forming ability were tested. CONCLUSION: Uropathogenic microorganisms infecting renal calculi and forming biofilms on them not only support chronic infection by increased resistance to therapy but also facilitate novel lithogenesis.

[Persistence of Burkholderia cenocepacia bacteria in vivo in dependence of their ability to form biofilms].

AIM: To select the most susceptible line of mice which allows to conduct comparative studies of infectious process caused by different strains of B. cepacia in order to explore correlation between ability to form biofilms and persistence of bacteria in organs of infected animals. MATERIALS AND METHODS: Strain B. cenocepacia 370, which is a clinical isolate, and its mutants with modified ability to form biofilms were used. Conditional microbiologic methods and biological models of intraperitoneal and intranasal inoculation of mice belonging to 4 lines: BALB/c, BLACK, I/St, and A/Sn derived in Central Institute of Tuberculosis were employed. Criteria of persistence was duration of isolation of different strains of bacteria from lungs and spleen of inoculated animals as well as number of CFU. RESULTS: The most susceptible line of mice which enables to conduct comparative studies of infectious process caused by Burkholderia species was determined. It was shown that even after intraperitoneal inoculation the agent was better preserved in lungs than in spleen that corresponds to natural localization of this infection. At any time of observation the number of cells of mutant strain, which is a superproducer of biofilms, isolated from organs of inoculated mice was 2 - 10 times higher than number of isolated cells of mutant, which do not produce biofilms. CONCLUSION: Correlation of more prolonged persistence of B. cenocepacia in organs of inoculated animals in vivo with ability of the agent to form biofilms determined in vitro is experimentally established. The susceptible line of mice which allows to conduct comparative studies of dynamics of infectious process caused by various strains of Burkholderia species was revealed. It was shown that irrespective from method of inoculation B. cepacia are able to continuously persist in organism of susceptible animals with lungs as a predominant localization.

[Electron microscopic study of Burkholderia cepacia biofilms].

Using the methods of transmission electron microscopy, the structure of the biofilms formed by the bacterium Burkholderia cepacia (clinical isolate and mutants with an increased and decreased ability to produce biofilms) were investigated. The biofilms were obtained on a liquid nutrient medium or on an abiotic surface (polystyrene). It has been demonstrated that the cultures of the studied strains differ in some morphological and functional characteristics. In biofilms, changes in the size and submicroscopic organization of all the components of bacterial cells occur. Staining biofilms with ruthenium red revealed the presence of exopolysaccharides in the intercellular space. The differences in the ultrastructure of bacterial films formed on nutrient medium and abiotic surfaces were demonstrated.

[Local tissue hypoxia consequence in the trophic venous ulceration in elderly patients].

This paper analyzes the data of examination and the results of the treatment of 25 patients aged over 80 years (mean age 68.5+/-7 years) suffering from varicosity with long nonhealing trophic ulcers of the distal limb segments (CVI CMP C6) and 20 patients of the same age groups with CVI CEAP stages 3-5. All patients with CVI underwent either full-scope phlebectomy (64.4%) or partial truncal phlebectomy (35.6%), in which the trunk of the greater saphenous vein was stripped up to the upper third of the leg. A TCM-3 outfit (RADIOmeter, Denmark) was employed to measure oxygen tension in limb tissues. Parameters of lipid peroxidation and antioxidant tissue defence were measured intraoperatively in the capillary blood of the fingers as well as in the venous blood withdrawn from the cubital vein and the greater saphenous vein in the lower third of the leg near ulcer). The data obtained evidenced that lipid peroxidation activity was most pronounced in the soft tissues of the lower third of the leg in the group of patients with remarkable chronic venous insufficiency without trophic venous ulcers (GVI CEAP C3-5) and was significantly depleted after formation of varicose ulcers (CVI CEAP C6) associated with remarkable tissue hypoxia (TepO2 1.7-7.0 mm Hg). In all patients with CVI, the syndrome of lipid peroxidation was associated with the lowering of antioxidant defence activity. Patients with trophic venous ulcers had the signs of active inflammation in the soft tissues of the leg. The data obtained in the course of the study made it possible to optimize the treatment policy for elderly patients with trophic venous ulcers. In addition to the lowering of venous hypertension, the treatment included correction of microcirculatory disorders related to local hypoxia. Of special importance was reperfusion attenuation in the postoperative period.

[Virulent properties of hospital strains of bacteria of the Burkholderia cepacia complex, isolated in hospitals of Moscow].

The results of the study of hospital strains of the B. cepacia complex, isolated in hospitals of Moscow, with the use of phenotypical and molecular-genetic methods are presented. The phenotypical methods made it possible to differentiate Russian strains and classify them with a group of genomovars (I, III, IV). As the result the epidemic importance of the strains with epidemic markers, having specific characteristics for every clinic, was determined. The detection of the collection of genes cepI and cepR in the strains made confirmed the epidemic importance of the stains which had, due to the regulatory "quorum sensing" (QS) system, the potential capacity for inducing infection and persisting in the patient's body. The presence of gene cepR in all strains and the absence of gene cepl in 33% of strains gave evidence to suggest that in some strains the activation of the production of pathogenicity factors required the presence of other bacteria having the fully developed QS system. Thus, the new complex approach with the use of phenotypical and molecular-genetic methods permits more precise identification of the source of hospital infection induced by the bacteria of the B. cepacia complex.

[Clinical strains of Burkholderia cepacia: characteristic and detection of the components in the quorum sensing regulatory system]. / Klinicheskie shtammy kompleksa Burkholderia cepacia: kharakteristika i vyiavlenie komponentov reguliatornoi sistemy quorum sensing.

Described in the paper are characteristics of B. cepacia clinical strains isolated from patients at Moscow hospitals. The strains were investigated for the presence of proteolytic, chitinolytic, hemolytic and lipase activities as well as for presence of components of the "Quorum sensing" gene activity regulatory system by using biological test-systems and in the polymerase chain reaction with primers to genes cepI and cepR.

[Nitric oxide in the lymphatic microvessel regulation]. / Uchastie oksida azota v reguliatsii funktsii limfaticheskikh mikrososudov.

Topical application of sodium nitroprusside on rat mesentery has a marked influence on lymph microvessels function. The drug causes a dilation of majority of lymphangions and decrease of the pacemaker activity of the vessel wall structures and valves. These changes do not lead to lymph stasis, and lymph flow velocity does not reduce. The non-selective inhibitor of NO synthase (N-nitro-L-arginine) intensifies vasomotions of lymph microvessels, modulates phasic contractile activity and increases lymph flow velocity. There is a time dependent dynamic of changes in action of N-nitro-L-arginine. During inhibition of endogenous NO synthesis the responses of lymph microvessels on sodium nitroprusside application are modified: the constriction of majority lymphangions and activation of valve work are observed.

[Influence of tumor necrosis factor on the growth of vegetative and nonculturable forms of Salmonella]. / Vliianie factora nekroza opukholi na razmnozhenie vegetativnykh i nekul'tiviruemykh form sal'monell.

The growth rate of the vegetative forms and the recultivation rate of the uncultivable forms of Salmonella isogenous strains, one of these strains carrying mutation in gene pqi, were studied. The multiplication rate of the vegetative and uncultivable forms of Salmonella control strain in the spleen of infected animals at the initial stages of the infectious process was shown (in vivo) to be considerably accelerated after the preliminary incubation of the culture with cytokine (tumor necrosis factor). The multiplication rate, in vivo and in vitro, of Salmonella vegetative and uncultivable forms with mutation in gene pqi did not change after the incubation of the cells with cytokine, which is indicative of an important role played by the product of this gene in the process of the interaction of bacteria with cytokines. The full nucleotide sequence Salmonella gene pqi was determined.

Changes in lymph microcirculation during pathological stress.

Stress caused by immobilization and acoustic stimulation produces considerable changes in lymph microcirculation. These changes manifested in dilation of microvessels, enhanced phasic contractile activity and its abnormal pattern, accelerated lymph flow, and recruitment of new lymphangions in drainage functions.

[Functional organization of lymphatic microvessels of the rat mesentery]. / Funltsional'naia organizatsiia limfaticheskikh mikrososudov bryzheiki krysy.

Lymph flow as measured with biomicroscopic and speckle-interferometrical techniques was very unstable and its rate did not depend on lymphangion diameter in the rat mesentery. A correlation was found between amount of cells in the lymph flow and other indices of lymphangion functions. All microvessels with phasic contractions and/or working valve were revealed as having a lymph flow. The phasic activity and the valve work correlated with each other. The higher the amplitude and frequency of the phasic activity and the longer the contraction cycle, the higher was the rate of the valve work.

[Genital tuberculosis and surgical treatment of it]. / Genital'nyi tuberkulez i ego khirurgicheskoe lechenie.

The paper presents the specific features of diagnosis and clinical course of genital tuberculosis under the present conditions. It shows it expedient to make a differential diagnosis of genital tuberculosis in 2 steps: 1) the use of the routine examination and 2) the study of specific antituberculous immunity. Genital tuberculosis risk groups to be obligatorily examined at a specialized tuberculosis control facility are identified. The specific features of the clinical course of genital tuberculosis, such as aptness to exudative processes, a combination with extragenital tuberculosis and hormonally depended gynecological diseases are noted. Indications for and efficiency of endosurgical interventions in genital tuberculosis are shown.

[The baroreflex component in the mechanism of the hypotensive action of atrial peptides]. / Baroreflektornyi komponent v mekhanizme gipotenzivnogo deistviia predserdnykh peptidov.

Changes in the arterial pressure (AP), heart rate (HR) and sensitivity of chronotropic baroreceptor reflex (SBR) evoked by i. v. infusion of alpha-ANP, AP-III, AP-II, and AP-I (1 mg/kg/min) were studied in alert Wistar rats. The SBR was tested by infusion of phenylephrine or sodium nitroprusside. All the ANP analogues, except AP-1, were shown to decrease the AP and evoke differential changes in the SBR. The SBR evoked by phenylephrine was increased by 60-80%, whereas SBR evoked by sodium nitroprusside was decreased by 30-60%. The specific changes in tachycardic and bradycardic components of the baroreceptor reflex after ANP analogues infusions seem to reflect participation of the baroreceptor reflex in the mechanism of ANP-induced hypotension.

[The role of plasmids in the regulation of delta-endotoxin synthesis in Bacillus thuringiensis H14]. / Rol' plazmid v reguliatsii sinteza delta-éndotoksina Bacillus thuringiensis H14.

The role of plasmids in regulation of delta-endotoxin synthesis by Bacillus thuringiensis H14 was studied. The derivatives of strain Is-1 H14 containing a 4Md plasmid integrated into the chromosome synthesize small crystals and are not toxic for the gnat larvae. The transceptional transfer into this strain of a plasmid coding for crystal synthesis from the strain 69-6 serotype H5 results in restoration of insecticidal activity to the level of the parental strain Is-1. Transcipients activity is increased 10-15 fold in case of 4Md plasmid excision from the chromosome and autonomous functioning. Evidently, 4Md plasmid from the strain Is-1 as well as a plasmid coding for crystal synthesis from the strain 69-6 contains the regulatory elements participating in the expression of crystalline protein genes localized on other plasmids. The existence of two cellular regulatory groups is supposed to result in the significant increase in crystalline protein synthesis.