[Immune response to lower urinary tract infections].

Urinary tract infections are one of the most common and widespread infectious diseases. A certain role in etiopathogenesis may play genetic predisposition, as well as a decrease in antiadhesive properties and an increase in urothelium permeability due to incompetence of bladder glycosaminoglycan layer. The prevalence of infectious diseases increases significantly with age, as well as in patients with chronic diseases. The introduction of modern biotechnology has allowed clinicians to greatly expand therapeutic armamentarium, while having a number of advantages, including minimal frequency of complications and adverse events, the possibility for long-term use, accessibility, and etc. Priority research areas include the study of toll-like receptors, which are transmembrane proteins that provide pathogen recognition and activate the immune response. The role of these receptors in the development of the immune response to urinary tract infections was evaluated in our study, which allows to predict the course of the disease and to increase treatment efficiency.

[Evaluation bacteriological analysis of urine in patients with long-term bladder drainage].

INTRODUCTION: The experience of comparative evaluation of the effectiveness of various types of urethral catheters in prevention of catheter-associated infection is described in this article. MATERIALS AND METHODS: The study included 69 patients treated at the CCH n.a. S.I. Spasokukotsky in the period from December 2017 to March 2018. The average age of patients was 67.5 years. In all patients, the bladder was drained by a two-way Foley catheter No. 16-18 Ch (100% silicone). In the 1st group (n=18), the bladder was drained with a standard urethral uncoated catheter, in the 2nd (n=16) - with a silver impregnated urethral catheter, in the 3rd (n=15) - with an urethral catheter coated with nitrofuran, in the 4th (n=20) urethral catheter with the possibility of controlled irrigation of the bladder and urethra with antiseptic solutions and (a new model of the urethral catheter developed during cooperative work of the Moscow State University of Medicine and Dentistry n.a. A.I. Evdokimov Urology Department and National Medical Research Center of Obstetrics, Gynecology and Perinatology n.a. V.I. Kulakov). RESULTS: The bladder was drained by Foley urethral catheter for more or equal 15 days. A microbiological study of urine (on the example of clinical isolates of conditionally pathogenic microorganisms) with preparation of an inoculum, inoculation of nutrient media, counting cultures of pathogenic bacteria and determining the sensitivity of pathogenic bacteria to antibiotics was carried out. The study showed the effectiveness of the new urethral catheter model in patients with long-term bladder drainage. CONCLUSIONS: Conducting multicenter studies evaluating the effectiveness of the proposed urethral catheter model with the inclusion of a larger number of patients will reduce the economic costs, associated with treating patients with prolonged bladder drainage in the long term by reducing the number of nosocomial infection cases and reducing postoperative day.

[POSITIVE EFFECTS OF MODERN IMMUNOCORRECTION ACCORDING TO THE ANALYSIS OF CYTOKINES AND SLPI IN PATIENTS WITH PYELONEPHRITIS].

The peculiarities of cytokines as compounds of immunogenesis are shown in the patients having acute (A) and chronic (Ch) pyelonephritis (PN). The combination of antibacterial therapy with Nukleinat and Galavit promotes the positive changes of cytokin-producing ability of immunocompetent cells and decrease in the level of proinflammation cytokines in blood and urine, secretory leucocyte protease inhibitor (SLPI) in urine. In children with PN and adult patients with diagnostically elevated titres of antibodies (IgG) to Herpes simplex virus, Cytomegalovirus are shown the positive effects of Kanephron® H and Proteflazidum, accordingly. Clinico-immunological effects of immunomodulators testify to the expediency of this usage in complex therapy with the aim to modulate the cytokine link of immunity for improvement of the effective treatment in APN and the protection against aggravation of kidney functioning in ChPN.

[The anti-viral activity of the complex glycyrrhizic acid-alpha-glutamyl-tryptophan against experimental lethal influenza infection in white mice caused by oseltamivir-resistant strain of the virus].

Influenza virus is a leading causing factor of infectious respiratory human pathology. The ability to implement the antigenic drift and development of drug resistance makes it important to develop novel anti-influenza drugs of wide spectrum of activity. In this work, we present the results of the study of the activity of a combination of glycyrrhizic acid with dipeptide alpha-glutamyl-tryptophan against oseltamivir-reistant strain of the virus Al Vladivostok/2/09 (H1 N1) on the model of lethal influenza infection in white mice. Application of Orvilax was shown to decrease the specific mortality of animals (index of protection 39-67% depending on the dose of the virus and drugs combination), to increase the mean day of death to 3.7-5.0 days and decrease the infectious titer of the virus in lung tissue to 1.3 Ig EID50/20 mg. The corresponding figures for the reference compound Tamiflu were 8-11%, 0.5-1.5 days, and 0.6 Ig EID50/20 mg. The use of Orvilax also led to reliable increase of the titers of interferon in the blood from 30.4 to 56.5 ME/mL. The results obtained allow the drug to be considered as a promising anti-influenza remedy that is active against the drug-resistant virus strains.

[Ingavirin treatment of experimental parainfluenza pneumonia in Syrian hamsters].

Parainfluenza viruses affect the upper respiratory tract in all age group patients, in children aged 6 months to 3 years in particular. The most urgent task is to design drugs to treat parainfluenza. This investigation studied the antiviral activity of Ingavirin (2-(imidazole-4-yl) ethanamide of pentandioic-1,5 acid) on a model of parainfluenza infection in Syrian hamsters. The drug was shown to restrict the infectious process in animal lung tissue. This restriction manifested itself as reductions in the infectious titer of parainfluenza virus in the lung tissue, in the degree of pulmonary edema and tissue cell infiltration, and in virus-specific lesion of bronchial epithelial cells. The in vitro experiments demonstrated the ability of Ingavirin to diminish the infective activity of viral descendants. The finding allows one to consider Ingavirin to be a promising antiviral agent that is active against parainfluenza infection in vivo.

[Effect of antiviral drugs with various mechanisms of action on morphogenesis of infection caused by extremely pathogenic influenza virus strains in animals].

The effect of meglumine salt of acridonoacetic acid (cycloferon) on the in vivo morphogenesis of influenza infection caused by viruses of different origin (avian, swine and human) and variable susceptibility to antivirals (rimantadine and oseltamivir) has been studied. The administration of cycloferon results in stimulation of the immune response, restriction of the foci of post-influenza pneumonia, and normalization of the structure of respiratory zones independently of the susceptibility or resistance of infectious virus to the drugs. Among virions formed in the lungs of cycloferon-treated mice, prevalence of irregular-shaped virions with defects of surface glycoproteins was observed. The data obtained suggest that cycloferon is a drug with the complex mechanism of activity.

[Antiviral activity of Ingavirin in experimental lethal influenza due to influenza virus B in albino mice].

The protective activity of Ingavirin against experimental infection caused by influenza B virus was studied on albino mice vs. Arbidol. Oral use of Ingavirin was shown to decrease the infectious titers of the virus in the animal lung tissue, to normalize the body weight dynamics, to lower the mortality and to increase the average lifespan vs. the placebo-treated animals. The activity of Ingavirin was higher than that of the reference drug. The results allowed to consider Ingavirin as a prospective agent for the treatment of influenza infection in humans.

[Protective activity of Ingavirin in experimental lethal influenza due to pandemic influenza virus A (H1N1)v in albino mice].

Despite obvious success in the vaccine development and chemotherapy of influenza, it remains a poorly controlled infection leading to emergence of new pandemic variants of the virus with high morbidity and mortality. We investigated the protective activity of Ingavirin against the lethal influenza A (H1N1) 2009 virus infection on albino mice. Oral use of Ingavirin resulted in sharp decreasing of the mortality (index of protection up to 57%), slight decreasing of the infectious titer of the virus in the lungs (up to 40-fold), normalizing of the body weight dynamics and the lung tissue structure vs. the placebo-treated control. The degree of the bronchial epithelium damage was also strongly decreased. The results allow to consider Ingavirin as an effective antiviral against the current pandemic influenza virus.

[Molecular cloning, expression, and characterization of human mini-antibodies against enterotoxin C1 of Staphylococcus aureus].

We describe here the cloning, expression, and production of specific single-chain antibodies (scFv) against the recombinant enterotoxin C1 of Staphylococcus aureus. High-affinity scFv were selected from the phage library of human mini antibodies; afterwards, the cells of E. coli trxA gor double mutant were infected with a product obtained by fusion of DNA encoding of these mini antibodies with the trxA gene to induce soluble scFv synthesis in cell cytoplasm. The scFv obtained displayed high enterotoxin C1 affinity. Analysis for cross reactivity showed that mini-antibodies interacted also with SEA- SEB-, SED-, SEE-, SEG-, and SEI-type enterotoxins, but they failed to interact with ricin, diphtheritic, and cholera toxins, or with both lethal and protective factors of the anthrax toxin. This property may be helpful in screening for staphylococcus enterotoxins.

[Production and characteristics of monoclonal antibodies to the diphtheria toxin].

Monoclonal antibodies to the diphtheria toxin were produced without cross reactivity with the thermolabile toxin (LT) from Escherichia coli; ricin; choleraic toxin; the SeA, SeB, SeE, SeI, and SeG toxins of staphylococcus; the lethal factor of the anthrax toxin; and the protective antigen of the anthrax toxin. A pair of antibodies for the quantitative determination of the diphtheria toxin in the sandwich variation of enzyme-linked immunosorbent assay (ELISA) was chosen. The determination limit of the toxin was 0.7 ng/ml in plate and 1.6 ng/ml in microchip ELISA. The presence of a secretion from the nasopharynx lavage did not decrease the sensitivity of the toxin determination by sandwich ELISA. The immunization of mice with the diphtheria toxin and with a conjugate of the diphtheria toxin with polystyrene microspheres demonstrated that the conjugate immunization resulted in the formation of hybridoma clones which produced antibodies only to the epitopes of the A fragment of the diphtheria toxin. The immunization with the native toxin caused the production of hybridoma clones which predominantly produced antibodies to the epitopes of the B fragment.

[Preparation and characterization of monoclonal antibodies to the cholera toxin].

Monoclonal antibodies to cholera toxin were obtained. They do not cross-react with the termolabile toxin (LT) of Escherichia coli, ricin, diphtherial toxin, staphylococcus enterotoxins of SEA, SEB, SEI, SEG, or the lethal factor and protective antigen of the anthrax toxin. Pairs of antibodies for the quantitative measurement of the cholera toxin in sandwich enzyme immunoassay (EIA) were selected. The detection limit of the toxin is 0.2 ng/ml for plate EIA and 0.44 ng/ml for microchip EIA. The presence of milk, broth, or surface water in the toxin samples does not reduce the sensitivity of EIA.

[Isolation and characterization of the ALP1 protease from Aspergillus fumigatus and its protein inhibitor from Physarium polycephalum].

It is known that Aspergillus fumigatus secretes a serine protease ALP1 of the subtilisin family in the presence of extracellular protein substrates. We found conditions of A. fumigatus culturing that provide a high ALP1 activity inside cells without induction by extracellular proteins. The identity of the properties of the secreted and intracellular enzymes was shown. A thermostable protein inhibitor of the ALP1 protease was isolated from the plasmodium of the myxomycete Physarum polycephalum. Its molecular mass is 32-33 kDa. The inhibitor inhibits the ALP1 protease activity with IC50 of 0.14 microM. This protein was also shown to be a less efficient inhibitor of the activity of HIV-1 protease (IC50 2.5 microM). The English version of the paper: Russian Journal of Bioorganic Chemistry, 2005, vol. 31, no. 3; see also http://www.maik.ru.

[Construction and molecular cloning of hybrid plasmids containing specific fragments of Escherichia coli DNA]. / Konstruirovanie i molekuliarnoe klonirovanie gibridnykh plazmid, soderzhashchikh opredelennye fragmenty DNK Escherichia coli.

In the paper a convenient procedure for the isolation of specific Eco RI-fragments of E. coli genome and their amplification on Km-resistance plasmid vector CKdelta11 is described. Plasmid CKdelta11 contains Col E1 replicon and has only one Eco RI site. The hybrid molecules were constructed in vitro using Eco RI-digestion followed by ligation. Then appropriated E. coli strain (polyauxotrophic strain E. coli K12 AB 2463) was transformed with ligated DNA mixture and hybrid plasmids, containing arg, leu, his and thr chromosomal markers were selected by molecular cloning and isolated from the obtained E. coli clones. The hybrid plasmids have two Eco RI sites and consist of one Eco RI-fragment of initial plasmid CKdelta11 and one Eco RI-fragment of El coli DNA. The method described allows to isolate and amplify on hybrid plasmids DNA fragments, containing any selectable genes or genes adjacent to the selectable ones.

[Physical mapping of the plasmid R205 and identification of a site responsible for increased UV-induced mutability of the bacterial host]. / Fizicheskoe kartirovanie plazmidy R205 i identifikatsiia raiona, otvetstvennogo za povyshenie UF-indutsiruemoi mutabil'nosti bakterii-khoziaina.

A physical map of the conjugative IncN plasmid R205 (56.1 kb) was constructed. The distribution of cleavage sites for investigated restriction enzymes is asymmetric. It was found that R205 suppresses the mutant phenotype of E. coli K12 umuC or umuD strains deficient in UV-induced mutagenesis. A mini-derivative of R205, designated pMU4 (15.1 kb) preserves the ability of the parent plasmid to increase the survival and induced mutagenesis of UV-irradiated host cells. A region of R205 located between 0 and 2.0 kb-on the plasmid map seems to contain information necessary for complementation of mutation in the host genes umuD/C, Hybridization between this region of pMU4 and plasmid pGW1700 bearing mucAB genes of pKM101 was observed.

[Transforming activity of Escherichia coli DNA fragments obtained following endonucleolytic splitting by EcoRI restrictase]. / Transformiruiushchaia aktivnost' fragmentov DNK Escherichia coli, poluchennykh posle éndonukleoliticheskogo rasshchepleniia restriktazoi EcoRI

Escherichia coli DNA with molecular weight of 20 - 10(6) daltons was digested by restriction endonuclease EcoR1, and the transforming activity of resctricts was studied. The transforming activity of restricts for two markers (Leu and Arg) was 2-5 fold increased, for two other markers (Thr and His) was not changed, and for one marker (Pro) was completely absent. The molecular weight of E. coli DNA restricts was 7,5-10(6) daltons. An increase and a decrease of the transforming activity for different markers appeared to be the result of two effects: 1) more efficient uptake of low molecular weight DNA into the cell and 2) the inactivation of markers as a result of location close to EcoR1 induced break.

[The antiviral activity of deitiforin in experimental para-influenza infection]. / Protivovirusnaia aktivnost' deitiforina pri éksperimental'noi paragrippoznoi infektsii.

Deitiforin in HEp-2 cell culture was shown to inhibit replication of the reference PIV-3 strain when administered 1 hour before virus inoculation. The most marked effect of the drug was observed in the first 4 days of observation. In experimental newborn mice infected with parainfluenza virus 3 deitiforin protected the animals from developing the infection. In humans given deitiforin reaction to vaccination was found to develop 5-6 times more rarely than in the control group, PIV-3 could be isolated twice as rarely, and a diagnostically significant rise of specific antibody levels was observed less frequently.

[Involvement of the afterbirth in influenza]. / O porazhenii posleda pri grippe.

Light- and fluorescence-microscopy examinations of 186 placentas were carried out, with concurrent determination of placental interferon, isolation of influenza virus, and serologic tests of maternal and fetal blood, and amniotic fluid. In 32 of the cases, placentitis caused by serotype A or B of influenza virus was present, characterized by hyperplasia and subsequent destruction of amniotic cells, trophoblast, decidual cells, and vascular endothelium, by the presence of influenza antigens, fuchsinophilic cytoplasmic inclusions in the affected cells, and lymphoid infiltrates, and/or by circulatory disturbances. In addition, an interferon possessing properties of viral, immune, and placental interferon was detected in a number of placentas. A or B influenza virus was isolated from 3 placentas. Diagnostic titers of anti-influenza antibody occurred in fetal blood and amniotic fluid samples. The present results indicate that influenza virus may persist and replicate in placental cells.

[Characterization of secondary immune deficiency in patients with chronic glomerulonephritis]. / Kharakteristika vtorichnogo immunodefitsita u bol'nykh khronicheskim glomerulonefritom.

A study was made of the nature of secondary immuno-deficiency, peculiarities of disordered function of regulatory T-lymphocytes (helpers and suppressors), and their interrelationships in different clinical variants of chronic glomerulonephritis (CGN). Altogether 94 CGN patients were examined, of them 34 were with urinary syndrome and 60 with nephrotic syndrome. With relation to a clinical variant of disease the absolute amount of T-lymphocytes was significantly lowered, the amount and function of B-lymphocytes showed a tendency to an increase. Function of nonspecific Concanavalin A-induced T-suppressors using PHA as a mitogenic indicator (T-T-suppression) was insignificantly raised, and when lipopolysaccharide (T-B-suppression) was used it was considerably suppressed. T-lymphocyte helper activity in CGN patients was on an increase.

[Activity of non-specific T-suppressors regulating the proliferation of B- and T-lymphocytes in patients with fibroadenomatosis, breast cancer and stomach cancer]. / Aktivnost' nespetsificheskikh T-supressorov, kontroliruiushchikh proliferatsiiu B- i T-limfotsitov, u bol'nykh fibroadenomatozom, rakom molochnoi zhelezy i zheludka.

A decrease in proliferative rate of blood-circulating lymphocytes in response to LPS and PHA was registered in patients with fibroadenomatosis and cancer of the breast and stomach cancer. The said cells preincubated with Concanavalin A showed a weak inhibitory action on B-cells. The inhibition of T-cell proliferation by lymphocytes either remained unchanged or became less apparent in stage II breast cancer and slightly increased in stage III gastric cancer. Since no correlation was established between proliferative levels of T- and B-lymphocytes, two separate subpopulations of non-specific lymphocytes (T-T and T-B) were suggested.